Abstract:Species of Microtus fortis, are always associated with rivers, flooded areas and wetland, and it is considered a good swimmer. To understand the adaptability of M. fortis to the aquatic movement, we observed and tested their swimming behavior, waterproof property of fur and ability in the laboratory and got the following results: M. fortis was able to swim on the water and underwater, but mainly swimed on the surface (Fig. 1), their swimming speed was about 0.37 m/s. The propulsion in water was maximized through a drag-based paddling motion. Their fur was waterproof that not only providing buoyancy, but also supporting they swimming longer time. They could swim as long as 5 h at 20 ℃ in this study. The waterproof quality of their fur was better in females than in males (P < 0.05), therefore female owned a bettter swimming ability (P < 0.05, Fig. 3). The long-term swimming ability of the vole is mainly determined by its fur waterproof performance (P < 0.01, R2 = 0.682, Table 3). At the same time, the resistance of fur to water had a negative correlation with their age and weight (P < 0.01, R2 = 0.805, Table 3). These results indicated M. fortis has well-waterproof fur and excellent long-term swimming ability. This species has evaluated adaptation to the aquatic movement.

Abstract:The Tibetan Plateau Toad (Bufo tibetanus) lives in the plateau regions at 2 400﹣4 300 m above sea level. This is the first report to analyze the call characteristics and auditory sensitivity of male toad. Calls of 6 individuals were recorded by directional microphone connected to a digital audio recorder. Auditory sensitivity of 13 males was measured by auditory brainstem response (ABR). The waveform and spectrogram of advertisement calls were drawn using Praat software (Fig. 1). The acoustic properties were obtained using Adobe Audition software. The advertisement call of male toad consists of a single note with a dominant frequency of 1 150 ± 99 Hz. ABR to tone pips were characterized by valley-peak waveforms (Fig. 2). The audiogram shows that the most sensitive frequency region is 1.4﹣2.0 kHz, but the ABR thresholds are above 70 dB across 0.6﹣6.0 kHz which means that this toad has a poor auditory sensitivity compared with other species (Fig. 3). The power spectra of the male call overlap the audiogram at 1.0﹣1.4 kHz, which supported the matched filter hypothesis, although the best auditory sensitive frequency (1.6 kHz) is slightly higher than the dominant frequency of the male call.

Abstract:Daya Bay is a provincial aquatic resources nature reserve, which is not only rich in biological resources, but also an important spawning and feeding ground for many economic fishes (Fig. 1). In order to protect the fishery resources of Daya Bay, four otter trawl surveys of fishery resources were conducted in April (spring), August (summer), October (autumn) and December (winter). Combining distance to the nearest mainland, distance to the nearest larger island, water depth, water temperature, salinity, chlorophyll a, total nitrogen, total phosphorus, maximum standard length of fish and catch per unit of effort, we discussed the nested patterns of fish assemblages in Daya Bay and its influencing factors (Appendix, Table 1). The result showed that the community composition of fishes in Daya Bay was significantly nested. The maximum standard length was significantly associated with their nested matrix ranks (Table 3), indicated that selective colonization was the main driver of nestedness of fishes in Daya Bay. All year round, the fish in Daya Bay tend to choose to inhabit the sea areas of Yangmeikeng and northern Dalajia Island (Table 2). This suggested that the sea areas of Yangmeikeng and northern Dalajia Island were the main distribution areas of fish. The Yangmeikeng was the main fattening area of fish, and the sea area of northern Dalajia Island was the fish spawning ground, both sea areas should pay prior attention to protection and management, moreover, both were the most suitable sites for fishery stock enhancement.

Abstract:Chemical signals are generally applied by spiders for intra- and interspecific communication. It is probably the first way that brought female and male spiders together for reproduction. In this study, choice preferences of adults and spiderlings in the funnel-web spider Pireneitega luctuosa to conspecific webs and body volatiles were tested using the two-choice arena system (Fig. 1). The Chi-square test was applied to analyze the preferences of tested spiders in the two-choice experiments using software SPSS 19.0 and Origin Pro 8. The results showed as the following: ① Both of the males and females displayed significant preference to webs (P < 0.001, Fig. 2 & Fig. 3) and body volatiles (P < 0.05, Fig. 5 & Fig. 6) of opposite sex in P. luctuosa. They stayed longer and showed more probing and weaving behavior in the chambers which were occupied by webs or body volatiles from the opposite sex. ② Both of the males and females were repelled by the body volatiles released from conspecific spiderlings (P < 0.05, Fig. 5 & Fig. 6). They preferred to stay and weave their webs in the control chambers instead of the chambers occupied by body volatiles of conspecific spiderlings. ③ Spiderlings of P. luctuosa were attracted by webs waved by both conspecific spiderlings and males (P < 0.001, Fig. 4). They were also attracted by body volatiles released from conspecific spiderings (P < 0.05, Fig. 7). Our results suggest that spiderlings of P. luctuosa preferred to live close to each other, which is probably the critical factor to aggregated distribution of this spider species. Our study verified that webs and body volatiles elicit preference or repellence behavior of conspecifics in the funnel-web spider P. luctuosa, thus confirming that semiochemicals associating with spider webs and bodies are applied for intraspecific communication and behavior orientation in P. luctuosa.

Abstract:Although pond culture is the major culture method for swimming crab Portunus trituberculatus, the growth and gonadal development pattern of pond-reared males remain unclear. Firstly, this study was conducted to investigate the changes of the growth parameters, the overall appearance and composition of mature reproductive system of male P. trituberculatus. Then, the changes in testis and vas deferens histology, gonadosomatic index, testis index, vas deferens index, hepatosomatic index and the gonadal development pattern during the process of pond culture were described. The results showed that: (1) The highest growth rate and specific growth rate of body weight and carapace width of male P. trituberculatus were found during the period of July to August (Fig. 2, 3). Furthermore, a significantly positive correlation was found between body weight and carapace width for pond-reared male P. trituberculatus (Fig. 4). (2) The mature reproductive system of male P. trituberculatus consisted of external and internal parts: the external part included the penis and petasma consists, while the internal part mainly included testis and vas deferens (Fig. 5). (3) Based on the changes of morphology, histology, and indices of testis and vas deferens, the gonadal development process of P. trituberculatus could be divided into three stages. Spermatogenesis (stage Ⅰ): the dominant types of gametocytes in testis were spermatogonia and primary spermatocytes, the testis index of stage Ⅰ ranged from 0.07 to 0.16, and the vas deferens appeared transparent and was hard to be found (Fig. 6, 7). Spermatophore formation (stage Ⅱ): secondary spermatocytes and spermatids were the dominant types of gametocytes in testis (Fig. 6, 7), and the testis index of stage Ⅱ ranged from 0.10 to 0.51. The vas deferens was filled with a large amount of fluid and spermatophores, and the vas deferens index of stage Ⅱ ranged from 0.01 to 0.41. Mature stage (stage Ⅲ): the gametocytes in testis were mainly composed of spermatids and spermatozoa (Fig. 6, 7), and the testis index of stage Ⅲ ranged from 0.10 to 0.41. The vas deferens was further enlarged and the vas deferens index of stage Ⅲ ranged from 0.20 to 0.65. (4) During the gonadal maturation, the gonadosomatic index and vas deferens index increased significantly, while the testis index showed a trend of increasing first and then decreasing (Fig. 8, 9). Furthermore, there was no significant correlation between gonadosomatic index and hepatosomatic index. (5) The gonadal development of pond-reared male P. trituberculatus was not synchronous. In July, the gonadal development of pond-reared male P. trituberculatus was mainly in stage Ⅰ (Fig. 11). At the end of August, the percentage of males reached stage Ⅱ was 76% (Fig. 11). At the end of September, the percentage of males reached stage Ⅲ was 47% (Fig. 11). From mid-late October to December, the gonadal development of pond-reared male P. trituberculatus was mainly in stage Ⅲ (Fig. 11).

Abstract:In October 2017, some Craspedacustas were discovered in Fuquan County, Guizhou province. In order to determine its taxonomy, we first observed and measured its morphological structures，and then amplified and sequenced its 18S rRNA, ITS and COI by PCR and DNA sequencing,, then finally compared its 18S rRNA, ITS and COI gene sequences with other Craspedacustas’ gene sequences in GenBank. The results showed that the umbrella diameter, level of tentacles, shape of statocyst and other morphology structures of Craspedacusta in Fuquan were highly similar to those of Craspedacusta sowerbyi (Table 1 and Table 5); the 18S rRNA and COI sequences of Craspedacusta in Fuquan was highly similar to those of C. sowerbyi (KY077287.1, MF000493.1), and the similarity rates were 100%, 99% respectively; the ITS sequences of Craspedacusta in Fuquan was highly similar to that of C. ziguiensis (AY513637.1), and the similarity rate was 99%. The genetic distances of 18s rRNA, ITS, COI gene sequences between Craspedacusta in Fuquan and C. sowerbyi were 0.001, 0.023, 0.012 respectively (Table 2﹣4). Based on 18s rRNA and COI gene region，the Neighbor-Joining (NJ) trees showed that Craspedacusta in Fuquan was the same species with C. sowerbyi; Based on ITS gene region，the NJ trees showed that Craspedacusta in Fuquan was the same species with C. ziguiensis (Fig. 6﹣8). Taken consideration of all the results of morphological observation and molecular identification, we determined that the Craspedacusta in Fuquan, Guizhou province was C. sowerbyi. We discussed some standards of morphological structure index of Craspedacusta, the possibility of using difference between umbrella diameters as a new classification standard in morphological taxonomy and give some opinions on methods for analyzing morphological structure.

Abstract:Tibetan sheep live in high altitude areas and possess good adaptability to high-altitude hypoxia. As the critical organs of spermatogenesis and sperm transportation, testicles are extremely sensitive to environment. The testicular and epididymal arterioles are regulated by the autonomic nervous system and control the blood flow in the entire microcirculation duct. Here, twenty testes of Tibetan sheep and Small-Tailed Han sheep were collected from their living areas including Qinghai and Gansu, respectively. The morphological characteristics of arterioles in testicular lobule and epididymides were investigated by using the blood perfusion technology and scanning electron microscopic method. The results demonstrated that arterioles of testicular lobule and epididymis exhibited a degree of curvature, and small arteries from the centripetal and centrifugal parts showed “dendritic” distribution（Fig. 1）. It was found that spiral of knot-like artery in Tibetan sheep showed tighter and more branches of small arteries than that in Small-Tailed Han sheep. The diameters of centripetal, Knot-like, centrifugal arteries and caput epididymal artery in Tibetan sheep were larger than those of Small-Tailed Han sheep (Table 1). Moreover, there were shallow imprints of the smooth muscles on the surface of testicular lobule and caput epididymis casts, but those in Small-Tailed Han sheep were deeper. Meanwhile there were more and intense precapillary constrictions in testicular lobule than those in Small-Tailed Han sheep (Fig. 2). The study suggested that the arteriole of testicular lobule and epididymis in Tibetan sheep contribute to vasoconstriction and sperm maturation, which became important anatomical characteristics for their adaptation to the plateau environment.

Abstract:We used light microscope to observe the histological characteristics of the dorsal, ventral skins and lungs of Bombina maxima, Megophrys glandulosa, Bufo gargarizans, Hyla gongshanensis, Rana chaochiaoensis, Dianrana pleuraden, Gynandropaa yunnanensis, Rhacophorus gongshanensis, Polypedates megacephalus, Microhyla fissipes, Kaloula verrucosa (Table 1). The results showed that the blood vessels in the back and the abdomen were interlaced into reticular veins. The morphological characteristics of dorsal skins, abdominal skins and lungs of these species were different (Table 2). The skin consisted of the epidermis and dermis (Table 3). Except for Hyla gongshanensis, the remaining 10 species had lamina calcarea distributed between the stratum spongiosum and stratum compactum of the dermis (Fig. 1). The pigment cells were located in the stratum spongiosum, with the dorsal pigment layer more developed. The alveolar sac, shaped hollow with dense blood vessels (Fig. 3), were divided into the right and left lobules of the same size. The lung was composed of pulmonary cyst wall, septum (Table 4), capillary and pulmonary alveoli. The pulmonary cyst wall consisted of the pleural layer, the middle layer and the inner layer: the pleural layer was composed of flat cells; the middle layer was composed of connective tissue; the inner layer was composed of pulmonary alveoli and capillaries; and the septum was composed of capillaries and connective tissues (Fig. 4). In these 11 species, individuals with more developed lungs tended to have reduced mucous glands in the skin structure. Individuals with more developed lungs had reduced mucous glands in their skin structure, and their histological features showed greater adaptation to the environment and less influence from phylogenetic relationships.

Abstract:At present there is no research on the establishment of Chinese zokor’s (Eospalax fontanierii) fibroblast cell lines and their biological characteristics. In this study, we selected 9 kinds of the tissues of Chinese Zokor, a representative animal genetic resource in the Mongolian Plateau, for the experiment. Fibroblast cell lines were successfully established from three tissues, including trachea, lung and xiphoid cartilage, and their biological characteristics were analyzed. The attachment rate and survival rate of the cells were calculated before cryopreservation and after resuscitation, and their chromosome karyotypes were analyzed. The fibroblast-like cells derived from primary culture of trachea, lung and xiphoid cartilage tissues were found in the third or fourth day of culture, and there was more than 90% confluence of adherent cells on the 11th, 16th and 17th day of culture, respectively. The morphology of these three kinds of somatic cells showed fibroblast characteristics (Fig. 1﹣4). The attachment ability of the trachea fibroblast cells was the greatest, and the attachment rate was 98.10% when cultured for 24 h, and the attachment ability of the lung fibroblast cells and xiphoid cartilage fibroblast cells were 95.28% and 94.88% respectively (Fig. 5﹣7). The analysis of the growth curve of fibroblast cells from three sources showed that the proliferative capacity of tracheal fibroblast cells was the best, the pulmonary fibroblast cells were the second and the xiphoid chondro fibroblast cells were the weakest. Tracheal fibroblast cells and pulmonary fibroblast cells entered the logarithmic growth stage (Fig. 8, Fig. 9) on the 6th to 7th day after inoculation. Xiphoid chondro fibroblast cells entered the logarithmic growth stage on day 2﹣3 (Fig. 10). The survival rate of cryopreserved fibroblasts from three different tissue sources decreased significantly. The growth curves of three fibroblasts are all in an “S” type. The multiplication capacity of the trachea fibroblast cells was the strongest, and the maximum proliferation number was 2.435 × 104 per well of a 24-well plate; the maximum proliferation numbers of lung fibroblast cells and xiphoid cartilage fibroblast cells were 1.813 × 104 and 1.521 × 104, respectively. The results of karyotype analysis showed that the chromosome number of fibroblast was 2n = 62 in Chinese Zokor (Fig. 11). To sum up, this study successfully established the fibroblast cells lines from Chinese zokor. The basic biological characteristics of fibroblast cells in this species were revealed, which provided basis for further studying molecular biology and physiological mechanism of Chinese zokor to adapt to the low oxygen and high carbon dioxide tunnel habitat, and provided important experimental materials and references for the further study of its heredity and species evolution.

Abstract:Human cytomegalovirus (HCMV) is popularly prevalent and extremely harmful, which has seriously hindered the researches on its pathogenic mechanism, drugs and vaccines, owing to restriction of species specificity and lacking animal models. In this study, we have explored the death of primary dermal fibroblasts (TSDF) isolated from the Chinese tree shrew (Tupaia belangeri) that infected with HCMV-Towne. The cytopathic effect and cellular death after infection were observed, and cell viability was measured using CCK (cell counting kit). The differences in transcriptional levels of apoptosis-related factor genes bax, bcl-2, and an unfolded protein response (UPR) relevant factor genes chop, atf4, xbp1s were detected using qRT-PCR. Western blot was also used to detect principal viral proteins IE1 and UL44 as well as apoptosis-related factors including Bax, Caspase-9, Caspase-3, and PARP. Additionally, apoptosis was detected by AV-PI double staining. The results showed that the cytopathic effect and cellular death gradually worsen as the infection progressed (Fig.1 a?e), accompanying with significant decrease of cell viability (Fig. 2c, P < 0.001). Moreover infection caused up-regulation of the transcription levels of bax, bcl-2, chop, atf4, xbp1s, and the bax and bcl-2 transcription levels showed a significant antagonistic trend (Fig. 3). Meanwhile, protein expression and progressive activation of Bax, Caspase-9, Caspase-3, and PARP were also up-regulated. Nevertheless viral proteins IE1 and UL44 were up-regulatied to the end of a complete virus replication cycle (Fig. 4). In summary, our study indicates that HCMV can induce apoptosis through cross-species infecting the tree shrew primary dermal fibroblasts, which is closely related to endoplasmic reticulum and mitochondria.

Abstract:Coilia nasus is an anadromous fish species with commercial importance and high market value in China, and it has a potential for aquaculture. In order to understand the value of changes in fatty acid compositions during embryonic and early post-embryonic development of C. nasus, the fatty acid compositions and contents at different developmental stages (embryos at 7﹣9 h of fertilization, newly hatched larvae at the age of 0-day, larvae at the age of 3-day, and larvae at the age of 5-day before feeding ) were collected and analyzed by biochemical analysis methods. The experimental data were statistically analyzed with variance analysis. The results showed that the total lipid percent contents of C. nasus larvae at different developmental stages were high (53.10%﹣60.97%, Table 1), while decreased significantly with ontogenesis (P < 0.05, Table 1). The individual total lipid percent contents decreased sharply with ontogenesis (P < 0.05), and the value dropped dramatically from 43.62 μg/ind in embryos to 16.27 μg/ind in 5-day-old larvae (Table 1); while, the moisture increased significantly with ontogenesis (P < 0.05, Table 1). A total of 6 saturated fatty acids (SFA), 4 mono-unsaturated fatty acids (MUFA), and 8 poly-unsaturated fatty acids (PUFA) were found in the dry sample at different developmental stages (Table 2). The C18:1n9c content of C. nasus was the highest and had an absolute advantage at different developmental stages (50.39%﹣57.00%, P < 0.05, Table 2), the C16:1 content was rich and stable (13.77%﹣14.24%, P > 0.05, Table 2), the C16:0 content was also rich (7.45%﹣9.15%) (Table 2). Simultaneously, the MUFA had an absolute advantage (65.14%﹣72.26%), the ratio of n-3 series poly-unsaturated fatty acids (∑n3PUFA)/ n-6 series poly-unsaturated fatty acids (∑n6PUFA) was low (1.78﹣2.38) (Table 2). During the incubation period, the degree of actual reduction of MUFA was higher, especially in C18:1n9c (13.21 μg/ind and 55.49%) and C16:1 (3.30 μg/ind and 53.12%)(Fig. 2), while, the degree of actual reduction of C20:5n3 (eicosapentaenoic acid, EPA) + C22:6n3 (docosahexenoic acid, DHA) was lower (1.44 μg/ind and 38.41%), especially in DHA (0.95 μg/ind and 36.52%, Fig. 2). After hatch, at the endogenous feeding stage, the utilization rate of MUFA was lower (1.94 μg/ind and 14.17%), especially in C18:1n9c (13.21 μg/ind and 12.41%) and C16:1 (0.63 μg/ind and 21.81%, Fig. 3), while the utilization rate of EPA + DHA was higher (1.04 μg/ind and 45.10%)，especially in DHA (0.71 μg/ind and 42.61%, Fig. 3). Therefore, EPA+DHA are preserved by priority during the C. nasus embryo incubation period, EPA + DHA (especially DHA) are largely consumed before the C. nasus larvae feeding, which is close to the fatty acids utilization characteristics of freshwater fish. In antepartum and postpartum cultivation of C. nasus brood stock, in order to improve the nutrition accumulation of brood stock and the recovery of physiological function of postpartum brood stock, diets enriched with the MUFA (especially C18:1 and C16:1) is suggested. In early larval breeding of C. nasus, in order to improve the survival rate of larvae, diets enriched with DHA and EPA (e.g., porphyridiophyceae and yolk) is also suggested.

Abstract:Oratosquilla oratoria is a common economic marine crustacean in China, which is known for its umami taste and high nutritional value. Due to offshore environment destruction and overfishing, the capture production of O. oratoria no longer meet the market demand, making it urgently important to pay closer attention to artificial culture. However, the pond culture of O. oratoria always showed defects in growth rate and gonadal development, and thus promoting its gonadal development had become a key technical problem in artificial culture. To solve this problem, this study investigated effects of different diet and water temperature on the survival and growth of O. oratoria during the overwintering period. In total 600 individuals (initial body weight: 21.99 ± 4.86 g) were used for this study, and sixteen samples (8 males and 8 females) were randomly selected and anatomized for each group on the day 0th, 45th, 90th to explore the influences of two kind foods (content of clam and frozen fresh fish) under two different temperature named natural (7.7 ℃ to 13.6 ℃) and heated (17.1 ℃ to 20.9 ℃) on the survival rate, body length, body width, body weight, gonad index, muscle index, and total edible rate. Results showed that: (1) The survival rate of the water heated group significantly decreased compared to that of natural temperature group (P < 0.05), which implied water temperature was one of the most important factors that influenced the survival rate during the overwintering period; (2) The body length, body width, body weight and other biological indices of O. oratoria kept an increasing trend throughout the whole process, without any significant difference found between any each groups (P > 0.05), and the best feeding pattern on growth effects was group of frozen fresh fish at natural temperature, while the worst was group of clam at natural temperature; (3) the gonad development condition under natural water temperature was better than that under heating water temperature in winter, and the optimal gonad development condition was achieved in group of clams at natural water temperature, with the gonad index 14.53% for females and 7.69% for males; (4) The muscle index showed an increasing tendency during the whole process with the highest muscle index (31.75%) detected in group of frozen fresh fish at natural temperature. For the edible rate, the only finding of significant difference was between the groups of clams at natural temperature and at heating temperature, respectively (P < 0.05). The highest edible rate (39.96%) was achieved in group of clams at natural temperature, while the lowest (36.08%) was found in group of clams at heating temperature. (5) The interaction results of different diets and water temperatures showed that diets had significant effects on ovary index (P < 0.01). In general, natural water temperature, especially ranging from 10 ℃ to 13 ℃, could meet the basic requirements for the growth and fattening performance of O. oratoria in winter. Compared to frozen fresh fish, clam was more beneficial for the substance accumulation in gonad of O. oratoria.

Abstract:We conducted a survey on the composition of amphibians and reptiles in Taihang Mountains of the western Xingtai with the purpose to promote management more effectively from July and August, 2018 in the 5 sampling areas (Fig. 1). Totally, we surveyed 20 transects as long as 3 km with 10 meter width each and 8 point samples whose set at the location with rocks only. As the results, we recorded 18 species, 5 species of amphibians and 13 species of reptiles. Those species belongs to 12 genus, 2classes, 2 orders (Appendix). The amphibians and reptiles in this area are relatively poor both in species and quantity. However, the dominant species have obvious quantitative advantages, they distributed in the high altitude distribution. The composition of the fauna is dominantly by Palaearctic realm species. Among the 18 species, 8 are the key protected in the region of Hebei Province, one is listed as endangered species and 3 vulnerable species in the “Red List of China’s Vertebrates”.

Abstract:Distribution and morphological characteristics of mast cells in gastrointestinal tract of Tilapia Galilaea were studied by modified toluidine blue staining. The results showed that the mast cells scattered or concentrated in the lamina propria mucosa and submucosa, especially around small blood vessels (Fig. 1). Mast cells varied in their sizes and shapes. They were round, oval, fusiform or rhombic in shape, and their nuclei were in dark blue when stained with toluidine blue, and the granules were in purple red with varying shades of color (Fig. 2). Statistical analysis (Table 1) by one-way ANOVA of SPSS 24.0 showed that the number of mast cells in stomach (9.7 ± 1.9), pyloric caecum (8.3 ± 2.8), hindgut (8.3 ± 1.8), foregut (6.3 ± 1.9) and midgut (5.1 ± 1.8) of Tilapia Galilaea decreased in turn. The number of mast cells in gastric and pyloric caecum was significantly higher than that in foregut or midgut (P < 0.05), but there was no significant difference from that in hindgut, and the number of mast cells showed no significant difference in the foregut, midgut and hindgut. In conclusion, mast cells were mainly distributed in the lamina propria and tunica submucosa of various gastrointestinal tissues, with different sizes and shapes, and the number of mast cells decreased successively in the order of stomach, pyloric caecum, hindgut, foregut and midgut.

Abstract:The aim of this study is to establish a method for purifying and culturing cerebral microglial cells of tree shrew (Tupaia belangeri), providing new experimental materials for future research. Newborn tree shrews were used in this experiment. The cortex of cerebrum was isolated and disaggregated by trypsin digestion. The microglia cells were purified by three methods after culturing for 9﹣10 days: upright hand clapping, mild digestion with trypsin and constant temperature oscillation. Meanwhile，we also used differential attachment method to further purify cells. At last, the number of purified cells was counted using a cell counter. Purified microglial cells were labeled with specific marker CD11b and observed with fluorescence microscope. The results showed that microglial cells were in a resting state on the third day of isolation and culture, with irregular shapes such as spindle, rod and branch (Fig. 2). Purified microglial cells were positively labeled with specific marker CD11b as revealed by fluorescence microscopy (Fig. 3). Data statistics was performed using SPSS statistical software. Analysis of cellular immunofluorescence intensities and counting of cells purified by different purification methods showed that the cell yield obtained by the upright hand clapping method was significantly higher than that obtained by constant temperature oscillation method, and the positive cell rate was significantly higher than that obtained by mild trypsin digestion method (Fig. 4). Primary microglial cells of tree shrews with high yield and purity were obtained by upright hand clapping.

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