Objective: To observe the protection of Testudinis Carapax et Plastrum extracts (TCPE) on serum starvation-induced PC12 cell apoptosis and explore its mechanism. Methods: The PC12 apoptosis model was established by serum starvation for 3 d. The cells were randomly divided into four groups: control group, model group, low-dose and high-dose (3 and 30 μg/mL) TCPE groups. In the three days of the treatment, cell absorbance was determined by MTT, ratio of cell apoptosis was examined by Annexin V/PI double stain flow cytometry (FCM), Caspase-3, BMP4, BMPR-IA, and p-Smad1/5/8 signaling molecular expression were detected by Western blotting, and the anti-apoptotic effect of TCPE was observed after blocking BMPs signal pathway. Semi-quantitative analysis of bands was carried out by Bio-Rad Quantity One gel analysis system. Results: MTT and FCM analyses demonstrated that TCPE could increase PC12 cell viability and decrease their apoptotic ratios in a dose dependent manner. Western blotting results showed that TCPE could decrease Caspase-3 expression, promote the expression of BMP4, BMPR-IA, and p-Smad1/5/8. There was statistically significant difference between TCPE (3 and 30 μg/mL) groups and model group (P<0.05, P<0.01) in all above results. While TCPE had no effect on the expression of BMP2, BMP7, and BMPR-II. BMPR-IB hadn't been detected. The anti-apoptotic activity was partially mitigated by neutralizing BMP4 antibody. Conclusion: TCPE has the capacity to inhibit the apoptosis of PC12 induced by serum starvation in a dose dependent manner and its mechanism may be associated with partially activating and up-regulating the expression of BMP4 signaling pathway.

Objective To improve the teaching method of anatomy and explore the application of virtual simulation experimental teaching platform in systematic anatomy. Methods Tencent questionnaire platform was used to carry out the "Questionnaire on How to Learn Systematic Anatomy".A comparative study was conducted among 400 students majored in clinical, nursing, general practice and imaging of 2017 in a medical college, two classes from each specialty.A virtual simulation experimental teaching platform of anatomy only used in the experimental group.The teaching effect was evaluated by theory examination, experiment examination and investigation by questionnaires on students'satisfaction with the teaching. Results The theoretical scores, experimental scores and total scores of the experimental group were higher than those of the control group, and the difference was statistically significant (P<0. 0 5). From the corresponding classes of each group, the theoretical scores, experimental scores and total scores of the same professional experimental group were higher than the control group, and the difference was statistically significant (P < 0. 05). There was a significant positive linear correlation between the theoretical and experimental scores of each class(R2 = 0. 97, P<0.05).The satisfaction rate of the experimental group on the various indicators of teaching effectiveness ranged from 81.00% to 93.00%, and the difference between the two groups was statistically significant (P < 0. 0 5). Conclusion The application of virtual simulation experimental teaching platform can effectively improve the teaching quality of systematic anatomy.

Objective To mvestigate the effect of phospholated-ERKl/2 on NF-κB p65 expresson nn the substania nigra(SN) of l-msthyi-4-phenyi-l,2, 3, 6-tetrahydropyeidine (MPTP)-induced mouse model of Parkinson's disease(PD). Methods PD mouse model was induced by MPTP and the behavoor of mouse was observed. Immunohistochemistry and Western bloiting analysis were used to observe the changes in expression of tyrosine hydroxylase (TH), NF-κB p65 and p-ERKl/2 in the SN of midbrain. Meanwhite, the above changes were also observed after treatment with U0126, a specific inhibitor of ERK. Results 1 h after the third MPTP administration, there were much more p-ERKl/2 positive cells than NF-κB p65 positive cells in the SN. 24 h after the fifth injection of MPTP, NF-κB p65 positive cells were significantly increased and p-ERKl/2 positive celiswere decreased, accompanted by marked loss of TH positive neurons. The above changes were greatly alleviated in animais treated with U0l26. Conclusion ERKl/2 pathway may regulate NF-κB p65 activation in MPTP-induced mouse model of Parkinson's disease, which leads to loss of dopamine neurons.

Objective To investigate the expression and function of aquaporin-4(AQP4) in the olfactory system of mice. Methods The differences of AQP4 expression in olfaction system between wild-type and AQP4-null mice were studied by immunoblotting and immunofluorescence methods. The differences of mouse olfactory functions in the two groups were examined by two olfactory behavioral assays: the busied food pellet test and olfaction maze test, and the odorant-stimulated electroolfactogram (EOG) recording. Results The results of immunoblotting and immunofluorescence showed no AQP4 expression in the olfactory system in AQP4-null mice. Immunofluorescence result also indicated that AQP4 was mainly distributed in the membrane of support cells, duct cells of Bowmann's gland, basal cells of olfactory epithelium, membrane of Bowmann's gland epithelial cells, olfactory sheath cells surrounding the olfactory bundles, and the membrane of cells in the olfactory bundle layer and glomeruler layer. The results of olfactory behavioral assay were significantly different between the two groups at all time points tested in both the olfaction maze test and the buried food pellet test (P<0.05). It was showed that the EOGs under different pressures of saturated trimethylamine had a similar shape in both groups, and the amplitude of EOGs increased with the increase of pressure. While under the same pressure, the EOG amplitude of AQP4-null mice was significantly lower than that of wild-type mice(P<0.05). Conclusion AQP4 is widely distributed in the olfactory system of mice, including the olfactory mucosa, olfactory nerve, and olfactory bulb, which can protect the olfactory neural bundle and facilitate neural signal transfer.

OBJECTIVE: To determine the cardiovascular actions of salusin-α in the caudal ventrolateral medulla (CVLM) in anesthetized rats. METHODS: Sixty-one anesthetic male SD rats were employed in the present study. The dose-dependant effects of salusin-α (0.04-4 pmol) on blood pressure and heart rate in the CVLM were determined by unilateral microinjection of salusin-α or artificial cerebrospinal fluid (aCSF) into the CVLM in 25 rats. In the other 36 rats, a CSF, kynurenic acid (Kyn), atropine, hexametho-nium (Hex) in the CVLM or a CSF/muscimol in the RVLM were given in advance to determine the mechanism of the cardiovascular actions of intra-CVLM salusin-α. RESULTS: Unilateral microinjection of salusin-a into the CVLM produced a dose-dependent hypotension and bradycardia. Prior administration of Kyn (1 nmol) or Hex (120 pmol) into the CVLM did not alter the hypotension and bradycardia induced by intra-CVLM salusin-α (P>0.05). But prior administration of atropine into the CVLM or pretreatment with muscimol within RVLM almost completely abolished the hypotension and bradycardia evoked by intra-CVLM salusin-α (P<0.05). CONCLUSION: Microinjection of salusin-α into the CVLM produces significantly hypotension and bradycardia, which probably originates from suppressing the activities of presympathetic neurons in the RVLM.

Objective: To establish the gene expression profile during early period of smooth muscle cell(SMC) proliferation in macroangiopathy in diabetic rats with spontaneous hypertension. Methods: Spontaneously hypertensive rats (SHR) were taken as control and a hypertensive diabetic rat model was established by streptozotocin (STZ) injection in SHR. The total RNAs were obtained from intima-media of thoracic aortas in 1-week, 2-week and 4-week hypertensive diabetic rats separately. The mRNAs from SHR and hypertensive diabetic rats were reversely transcribed into the cDNAs. The 2 probes were then hybridized to the cDNA microarray containing 4 096 full length genes from rats. After high-stringent washing, the cDNA microarray was scanned and analyzed by computer image analysis. Bioinformatic analysis of altered gene expression was performed to find out the possible genes related to diabetic macroangiopathy. Results: A total of 321 genes were screened out in 1-week hypertensive diabetic rats, with 160 genes down-regulated (ratio<0.5-fold) and 161 up-regulated (ratio>2-fold); 414 genes were screened out in 2-week hypertensive diabetic rats with 128 down-regulated and 176 up-regulated; and 403 genes were screened out in 4-week hypertensive diabetic rats with 202 down-regulated and 201 upregulated. The effective genes were divided into 8 groups by cluster analysis. Thirteen known genes were up-regulated in all 3 kinds of hypertensive diabetic rats, among which CD74, Irf1, GAP43 and CD36 had a steady increase and were related to cell proliferation. Furthermore, CYP2E1 gene was obviously up-regulated (ratio for the 1-week rats was 34. 54,2-week was 24. 82, and 3-week was 33.57). Conclusion: The gene expression profile based on cDNA micrioarray can be used for screening genes related with diabetic macroangiopathy. It is found that CYP2E1 might be an important gene responsible for proliferation of SMC in diabetic macroangiopathy.

Background: Studying the human embryonic and fetal organ systems development patterns and determining their quantitative indicators is of scientific and practical importance in medicine and health in every nation.
Distortions and pathologies during the development of the embryo are the causes of congenital disabilities. Among the congenital malformations, facial malformations are the 3rd place, including cleft lip and palate in 70% and Srouzon's syndrome in 30%. In addition, abnormalities due to changes in the size, shape, and position of the jaw are also mentioned in the 2021.04.21 issue of Morphology magazine in the study "Morphometric parameters of the bones of the skull and face during the development of newborns and fetuses". In our country, Ariuntuul G (2005) determined that cleft lip and cleft palate occur at 0.76/1000 or 1 in 1314 live births, while Ayanga G (2012) found that it occurs at 1 in 1072 live births or 0.93/1000. Moreover, the eye cup dimensions of Mongolian fetuses aged 16 36 weeks have a positive linear relationship with the gestational age determined using ultrasound by Nandintsetseg B (2015) et al. Compared with the other countries, the eyecup is slightly wider, and the outer edge distance is similar, whereas the inner edge distance is shorter. Purpose: To summarize research work and determine the embryonic development of bones involved in the formation of the face and facial parts, the period of bone formation, the point of ossification, and the period of formation. Methods: During fetal development, human organ systems grow and develop at different rates but in a particular relationship. This feature of growth and development is also clearly observed in the structure of the head and facial bones, and the results of researchers who have studied this aspect are selected in the articles. Results: Embryonic and fetal development of bone are clinically significant not only from the point of view of its morphogenesis but also from the point of view of congenital disabilities. Conclusion In the analysis of the sources, most of the works on the prenatal period of the development of the same body have studied the development of specific structures of the face and facial area, such as the palatine bones and nasal bones, or have generally covered the development of particular systems in the embryo and fetus, and face, there are relatively few works that show the entire dynamics of growth and development of facial bones.

Objective To explore the effect of the total saponins of Panax notoginseng (TSPN) on the expression of GFAP in hippocampus and brain water content in rats subjected global cerebral ischemia injury. Methods Using four-vessel occlusion method built the global cerebral ischemia model. Rats were divided into Sham group, vehicle group, and TSPN group. The rats in the TSPN group were administered TSPN intraperitoneally 30 min post-brain ischemia. The dose of TSPN (75 mg/kg) was suspended in 0.9% saline, once per day for days 1, 3, 7, and 14 after reperfusion. While rats in the vehicle group was treated with equal volume of 0.9% saline, one injection per day until the rats were sacrificed at either days 1, 3, 7, and 14 after brain ischemia. The brain water contentwas detected by dry-wet technique and GFAP expression in the dentate subgranular zone (SGZ) was assessed by immunohistochemistry. Moreover, immunofluorescence was applied to detect the GFAP/DCX in the SGZ, and western-blot was adopted to test the protein level of GFAP. Results The brain water content in the TSPN group was significantly lower than the vehicle group (P < 0.05); There was statistical difference in the GFAP+ cells density in SGZ at the 3th, 7th, 14th day in two groups (P < 0.05). Furthermore, compared with the vehicle group, the ratio of the GFAP/DCX to DCX in the SGZ at 7th, 14th d of TSPN group was significantly different (P < 0.05), and the protein level of GFAP on days 3, 7, 14 in the TSPN group was higher (P < 0.05). Conclusion TSPN could play a neuroprotective effect through promoting gliosis, neuroregeneration in the SGZ, and alleviating brain water content of rats following global cerebral ischemia.

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with Gastrodin on learning-memory ability and expression of silent information regulator 2 homologous protein 1(SIRT 1) and peroxisome proliferator activated receptor γ coactivator (PGC-1 ɑ) of hippocampal CA 1 region in Alzheimer's disease(AD) rats, so as to explore its mechanism under-lying improvement of AD. METHODS: Sixty male SD rats were randomly divided into normal control (normal), sham operation (sham), model, EA, Gastrodin and EA＋ Gastrodin groups (n＝10 in each). The AD model was established by intraperitoneal injection of D-Galactose (120 mg•kg－1•d－1) combined with bilateral hippocampal injection of β amyloid 1-40(Aβ 1-40). EA was applied at "Baihui"(GV 20), "Dazhui"(GV 14) and "Zusanli"(ST 36) for 30 min, once daily for 4 weeks. For rats of the Gastro-din group and EA＋ Gastrodin group, intraperitoneal injection of gastrodin(10 mg/kg) was conducted once daily for 4 weeks. Morris water maze tests were used to assess the rat's learning-memory ability. Nissl staining was used to assess the morphological changes of neurons in the hippocampal CA 1 area. The expression of SIRT 1 and PGC-1 ɑ of hippocampal CA 1 region was mea-sured by immunohistochemical staining. RESULTS: 1) Morris water maze tests showed that, compared with the normal and sham group, the escape latency was significantly prolonged (P<0.05), and the percentage of platform quadrant residence duration and the platform crossing times were considerably decreased in the model group (P<0.05). After the intervention, the escape latency was obviously shortened (P<0.05), and the percentage of platform quadrant residence duration and the platform crossing times were markedly increased in the EA, Gastrodin and EA＋Gastrodin groups relevant to the model group (P<0.05). 2) Nissl staining showed that, in comparison with the normal group or sham group, the number of cells in the hippocampal CA 1 area was decreased and the arrangement was disorganized in the model group. The number of cells in CA 1 area was relatively higher in the 3 treatment groups than in the model group. 3) The expression levels of SIRT 1 and PGC-1 ɑ proteins in the hippocampal CA 1 area were significantly down-regulated in the model group than in the normal and sham groups (P<0.05). After the intervention, the expression levels of SIRT 1 and PGC-1 ɑ in the EA, Gastrodin and EA＋Gastrodin groups were significantly up-regulated compared with the model group (P<0.05). The effects of EA＋Gastrodin were significantly superior to those of simple EA and simple Gastrodin in shortening the escape latency, up-regulating the expression levels of SIRT 1 and PGC-1 ɑ as well as in increasing the percentage of platform quadrant residence time and platform crossing times (P<0.05). CONCLUSION: Both EA and Gastrodin can improve the learning-memory ability of AD rats, which may be related to their effects in up-regulating the expression of SIRT 1 and PGC-1 ɑ and reducing neuronal injury in the CA 1 region of hippocampus, suggesting a protective role of EA on hippocampal neurons. The effect of EA combined with Gastrodin is markedly better than that of EA and Gastrodin alone.

Objective: To observe the localization and expression of dopamine receptors in olfactory bulb (OB) of rats and explore the effect of L-levodopa (L-DOPA) treatment on hyposmia in Parkinson' s disease (PD) rat model. Methods: Western blotting, immunohistochemistry and immunofluorescence were used to observe the expression and localization of dopamine receptors in the OB. PD rat model was established by bilateral 6-hydroxy dropamine(6-OHDA) injection to detect the effect of L-DOPA treatment on the hyposmia and the expression of glutamic decarboxylase (GAD) and brain derived neurotrophic factor (BNDF) of PD rats. Results Dl and D2 receptors were the major subtypes in the OB.D1 and D2 receptors were expressed by GAD positive Î3-aminobutyric acid (GABA) ergic neurons in the granule cell layer(GCL) which surrounded by tyrosine hydroxylase (TH) positive nerve fibers. The expression of TH in the GCL layer of PD rats decreased significantly (0. 05±0. 01 vs 0. 01±0. 00,P<0. 001). After L-DOPA treatment, the time of finding food balls in PD rats was significantly reduced [(624. 4±113.4)s vs (312. 4±79. 35) s, P<0.05]and the expression of BDNF in the OB was increased (0. 02±0. 01 vs 0. 07±0. 01, P<0. 01). Conclusion Dl and D2 are expressed in the GABAergic neurons in the GCL layer of OB. L-DOPA treatment alleviates the hyposmia of PD rats, which may be related to the D1-D2 receptor heteromeractivation and its downstream BDNF expression of GABAergic neurons.