Objective To establish the quality control method for Youqing Granules. Methods Qualitation discrimination of Rhizoma Cyperi and Raidix Paeoniae in this formula was identified by TLC. Quantitive analysis of Astragaloside A which is the effective element of the formula was adopted by RP-HPLC-ELSD. Result The specks in TLC were clear and easy to discriminate. The precision and reproducibility of the RP-HPLC-ELSD method were fine. In 1.032～5.16 ?g, the content of Astragaloside A had good linear relationship. The average recovery rate of sample was 99.0516 %, and RSD=1.05%. Conclusion The quality standard established on this study can control the quality of the products, the method is viable.

Objective To investigate the prevalence of Helicobacter hepaticus (H. hepaticus)infection in various species of mice from different regions of China in order to find the role of H. hepaticus in development of hepatitis, liver cancer and tumors in lower digestive tract in mice.Methods One hundred and fourteen mice, including C57BL/6 mice (n= 39), BABL/C mice (n=45),SCID mice (n=14) and C3H mice (n=18), were collected from different regions of China. The serum anti-H, hepaticus-IgG and fecal H. hepaticus antigen were determined by using ELISA. Polymerase chain reaction analysis (PCR) was used to screen Helicobacter genus-specific 16SrRNA and H. hepaticus species-specific 16SrRNA. The feces were cultured and identitied for Helicobacter infection in 114 mice. The H. he paticus infection was identified as one of above tests being positive.Results Of 114 mice, 25 (21.9%) mice were infected with Helicobacter species. The mice infected with H. hepaticus accounted for 44. 0% (11/25) with SCID and C3H mice in high prevelence.Meanwhile, the PCR examination revealed that the rest 56.0% (14/25) mice were infected with other Helicobacter species. Conclusion Besides H. hepaticus infection, the other Helicobacter species infections are also existed in China.

AIM:To determine four bioactive components in Tongda Granule ( Radix Paeoniae alba,Flos Carthami,Radix Scutellariale,etc. ),namely hydroxy safflower yellow A,peoniflorin,ferulic acid,and baicalin. METHODS:The isolation was performed on an Alltech Apollo C18 (250 mm ? 4. 6 mm,5 ?m) with the mobile phase consisting of methanol-0. 2% (w)phosphoric acid for gradient elution. The flow rate was 1. 0 mL/min and the column temperature was set at 30 ℃. RESULTS:This method was successfully applied to determining four bio-active components in Tongda Granule. The standard curves were linear over the ranges of 5. 20 -104 mg/L for hydroxy safflower yellow A(r =0. 999 1),25. 6 -512 mg/L for peoniflorin (r =0. 999 6),7. 72 -154. 4 mg/L for ferulic acid (r =0. 999 5),32. 5 -650 mg/L for baicalin (r = 0. 999 5),respectively. The average recoveries were 101. 1% ,99. 6% ,98. 9% and 101. 9% ,RSD were 2. 3% ,1. 8% ,2. 6% and 1. 3% ,respectively. CONCLUSION:The method is simple and can be used as a quality control method for Tongda Granule.

ObjectiveTo obtain stable animal models and observe Helicobacter hepaticas (Hh)colonization and pathological claracteristics,through infecting different mice strains with Hh.Methods SPF-class male BABL/c Cr,SCID/Cr and C57BL/6 Cr mice were inoculated 0.2 ml Hh standard strain ATCC51450 bacterial suspension (1 × 108CUF/ml),inoculated for 3 times with 48 hours intervals,the control group was fed with the same volume of PBS.Mice were executed at 4 weeks,8weeks and 16 weeks since last Hh inoculation,and mice esophagus,stomach,jejunum,ileum,cecum,colon,liver and pancreas tissue were taken for histopathology examination,Micro-aerobic bacteria isolation,culture and identification and Hh specific 16S rRNA gene amplification.Results The colonization rates of Hh in cecum after inoculated in BALB/c Cr mice and SCID/Cr mice at 4 weeks,8 weeks and 16 weeks were all 8/8,colonization rates in colon at 4 weeks,8 weeks and 16 weeks were 4/8,5/8,5/8 and 3/8,6/8,5/8 respectively,colonization rates in ileum and jejunum at 16 weeks were 1/8,colonization rates in liver at 8 weeks and 16 weeks were 2/8,3/8 and 2/8,2/8respectively.The colonization rates of Hh in cecum after inoculated in C57BL/6 Cr mice at 4 weeks,8weeks and 16 weeks were 1/8,2/8 and 2/8 respectively,colonization rates in colon at 8 weeks and 16weeks were 1/8,2/8 respectively.Compared with C57BL/6 Cr mice,the inflammatory changes in liver,cecum and colon were more significant in Hh infected BALB/c Cr and SCID/Cr mice (P＜0.01),and histological scores gradually increased as infection time extended (P＜0.05,P＜ 0.01 ).The histological scores were significantly higher in those with colon and liver Hh bacterial colonization than those without Hh bacterial colonization (P＜0.05).The histopathological score of cecal tissue was positively correlated with the density of Hh colonization.ConclusionDifferent mice strains are with different susceptibility to Hh,and better Hh infection model can be obtained in Hh inoculated BALB/c Cr and SCID/Cr mice.

BACKGROUND:Numerous clinical studies have confirmed that the microenvironment at a spinal cord injury site can be obviously improved through hyperbaric oxygen therapy; however, what effect does hyperbaric oxygen have on the microenvironment of the injured brain? OBJECTIVE:To investigate the effect of hyperbaric oxygen therapy on nerve regeneration microenvironment and the recovery of rat nerve function after focal cerebral infarction. METHODS:Rat models of focal cerebral infarction were established by occlusion of the middle cerebral artery and subjected to hyperbaric oxygen therapy. Sham group and model group were established as comparison. In the sham group, rat models of focal cerebral infarction were established but did not receive any treatment. Rats in the model group were placed in a hyperbaric oxygen therapy chamber but the pressure and oxygen concentration were not administered. RESULTS AND CONCLUSION:Compared with the model group, the score of rat limb function at 16 days after treatment and the expression of growth associated protein 43 in the rat cerebral infarcted area at postoperative 14 days were significantly increased , but infarct volume at postoperative 24 hours was al significantly decreased in the hyperbaric oxygen therapy group (alP < 0.05). These results confirmed that hyperbaric oxygen therapy can improve nerve regeneration microenvironment and promote the recovery of rat nerve function after focal cerebral infarction.

BACKGROUND:Studies have shown that human telomerase reverse transcriptase (hTERT) has the ability to enhance cel proliferation, maintain telomere length, prolonged cel life cultured in vitro. OBJECTIVE:To observe the effect of hTERT gene-modified bone marrow mesechymal stem cel transplantation on neural function recovery of rats with cerebral infarction. METHODS:Rat models of middle cerebral artery occlusion were established and randomized into model group, cel transplantation group and hTERT-modified cel transplantation group, with 20 rats in each group. Rats in the three groups were respectively injected via tail vein with 1 mL PBS, passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L) and hTERT-modified passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L), respectively. Modified neurological severity scores were determined before and after transplantation; RT-PCR and western blot assay were used to measure hTERT expression at gene and protein levels; TUNEL method was adopted to detect cel apoptosis in the brain. RESULTS AND CONCLUSION:hTERT-modified bone marrow mesenchymal stem cels had prolonged cel cycle, and with the increase in passage number, the cels showed good growth with no changes in morphology. The expressions of hTERT mRNA and protein were superior in the hTERT-modified cel transplantation group than the cel transplantation group, and there was a significant difference (P < 0.05). Modified neurological severity scores and number of apoptotic cels were decreased significantly in the hTERT-modified cel transplantation group compared with the other two groups (P < 0.05). These findings indicate that hTERT-modified bone marrow mesenchymal stem cels can promote neural functional recovery of rats with cerebral infarction.

Objective:To evaluate the expression of semaphorin 5B (SEMA5B) in gastric adenocarcinoma and its relationship with prognosis.Methods:In November 2019, the clinicopathological characteristics and SEMA5B mRNA expression data of 341 patients with gastric adenocarcinoma were collected through TCGA database. The relationship between SEMA5B expression in gastric adenocarcinoma tissues and clinical pathologic features and overall survival were analyzed. Gene Set Enrichment Analysis (GSEA) was used to analyze the signaling pathways regulated by SEMA5B.Results:The expression level of SEMA5B mRNA in 341 gastric adenocarcinoma tissues was 0.577±0.587, in adjacent normal tissues was 0.132±0.075, the difference was statistically significant ( P<0.001). The median survival time of 109 patients with high expression of SEMA5B mRNA was 14.5 months, 232 patients with low expression of SEMA5B mRNA was 17.9 months ( P=0.047). Univariate analysis showed that the expression of SEMA5B mRNA was correlated with histological grade and T stage ( P<0.05). The multivariate analysis revealed that age<65 years remained independently associated with overall survival, with a hazard ratio( HR) of 1.042 (95% CI: 1.021-1.064). The multivariate analysis revealed that high expression of SEMA5b mRNA remained independently associated with overall survival, with a HR of 1.195 (95% CI: 0.925-2.551). GSEA showed that malignant tumor signaling pathways ( P=0.008), MAPK signaling pathways ( P=0.047) and Notch signaling pathways ( P=0.029) were differentially enriched in SEMA5B highly expressed phenotype. Conclusions:SEMA5B expression may be a potential prognostic molecular marker for prognosis of GAC patients. Moreover, malignant tumor signaling pathway, MAPK signaling pathway and Notch signaling pathway may be the key pathway regulated by SEMA5B in GAC.

Objective:To evaluate the expression of semaphorin 5B (SEMA5B) in gastric adenocarcinoma and its relationship with prognosis.Methods:In November 2019, the clinicopathological characteristics and SEMA5B mRNA expression data of 341 patients with gastric adenocarcinoma were collected through TCGA database. The relationship between SEMA5B expression in gastric adenocarcinoma tissues and clinical pathologic features and overall survival were analyzed. Gene Set Enrichment Analysis (GSEA) was used to analyze the signaling pathways regulated by SEMA5B.Results:The expression level of SEMA5B mRNA in 341 gastric adenocarcinoma tissues was 0.577±0.587, in adjacent normal tissues was 0.132±0.075, the difference was statistically significant ( P<0.001). The median survival time of 109 patients with high expression of SEMA5B mRNA was 14.5 months, 232 patients with low expression of SEMA5B mRNA was 17.9 months ( P=0.047). Univariate analysis showed that the expression of SEMA5B mRNA was correlated with histological grade and T stage ( P<0.05). The multivariate analysis revealed that age<65 years remained independently associated with overall survival, with a hazard ratio( HR) of 1.042 (95% CI: 1.021-1.064). The multivariate analysis revealed that high expression of SEMA5b mRNA remained independently associated with overall survival, with a HR of 1.195 (95% CI: 0.925-2.551). GSEA showed that malignant tumor signaling pathways ( P=0.008), MAPK signaling pathways ( P=0.047) and Notch signaling pathways ( P=0.029) were differentially enriched in SEMA5B highly expressed phenotype. Conclusions:SEMA5B expression may be a potential prognostic molecular marker for prognosis of GAC patients. Moreover, malignant tumor signaling pathway, MAPK signaling pathway and Notch signaling pathway may be the key pathway regulated by SEMA5B in GAC.

Objective To investigate the effects of MCSO on physiological behavior and antioxidant index in D.melanogaster.Methods One-day-old wild type D.melanogaster was divided into control group,0.25％,0.5％,1％,2％and 4％dose groups,as well as male and female groups.The control group was exposed to the base medium,and each dose group was exposed to the MCSO medium added with 0.25％,0.5％,1％,2％and 4％concentrations,respectively.The optimal dosage concentration and time of administration were investigated by climbing experiment.Then the flies were divided into control group,model group and MCSO group.The model group was established by depriving the flies of sleep through repeated nocturnal light stimulation.Period of drug treatment,appetite test,negative geotaxis ability test,stress test,olfactory memory test,and sleep-wake rhythm detection were used to explore the effects of MCSO on their physiological behavior.The activities of super oxidase dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)were detected by enzyme-linked immunosorbent assay.Results MCSO enhanced the locomotory ability of 30-day-old D.melanogaster(P＜0.01),increased the activity of SOD and CAT(P＜0.01),and decreased the concentration of MDA(P＜0.01).Improve olfactory memory of senile fruit flies.After sleep deprivation,the night sleep time of female Drosophila model group was reduced(P＜0.05),and that of male Drosophila model group was reduced(P＜0.01).After feeding MCSO,the night sleep time of female drosophila model group was extended(P＜0.05),and that of male drosophila model group was extended(P＜0.01).Conclusions MCSO had a certain antioxidant effect,prolonging the sleep time and improving the olfactory memory of sleep-deprived Drosophila.

Insomnia has become a common central nervous system disease. At present, the pathogenesis of insomnia is not clear. Animal models can help us understand the pathogenesis of the disease and can be used in transformational medicine. Therefore, it is very necessary to establish an appropriate model of insomnia. Clinical data show that insomnia patients with high levels of thyroxine and often accompanied by cardiovascular problems, a common mechanism underlying all of these physiological disruptions is the sympathetic nervous system. Combined with the characteristics of chronic onset of clinical insomnia, an insomnia model induced by long-term intraperitoneal injection of thyroid hormone has been created in our laboratory. In this paper, the insomnia-like state of the model was evaluated based on three validity criteria. Face validity has been demonstrated in metabolism, the Morris water maze, electrocardiogram (ECG) and electroencephalogram (EEG). Structure validity has been proved by the results of targeted metabolomics. After treatment with diazepam, a commonly used clinical anti-insomnia drug, the above physiological and pathological disorders were reversed. The results of comprehensive analysis show that the established thyrotoxicosis-associated insomnia model meets the validity requirement to establish an appropriate animal model of insomnia. The model presented in this article might help to study pathogenetic mechanisms of clinical insomnia, as well as to test promising methods of insomnia treatment.