BACKGROUND: Diabetic retinopathy is the common complications of diabetes mellitus. Taurine is free aminoacid with the mostly abundant content in retina, and it is the necessary nutrition factor for regeneration and development of retina. Deficiency of taurine will cause structural and functional change of retina.OBJECTIVE: To probe into the effect of taurine on the expression of retinal glial fibrillary acidic protein (GFAP) mRNA DESIGN: A randomized and controlled observation SETTING: Third Military Medical University of Chinese PLA MATERIALS: This experiment was carried out at the Third Military Medical University of Chinese PLA from March 2003 to December 2003.Totally 54 closed group SD rats, with body mass of 250 g, were chosen.The successful diabetic rat model were randomly divided into 6 groups: diabetic group of 1 month, taurine-treated diabetic group of 1 month, diabetic group of 2 months, taurine-treated diabetic group of 2 months, diabetic group of 3 months, taurine-treated diabetic group of 3 months, with 9 rats in each group. Another 15 rats were chosen homeochronously as normal control group.METHODS: The animals were raised in separate cages. They were free to access to food and water, and fasted for 12 hours before experiment. Streptozotocin was used to induce diabetes mellitus .When Tes-Tape showed urine glucose reached more than (+++), and blood glucose concentration of venous blood from tail measured ＞16.7 mmol/L, models were established successfully. Expressions of GFAP mRNA and protein were measured with immunohistochemistry, RT-PCR, Western blotting and other technical methods. The effect of taurine on retinal neurons and glial cells in rats with diabetes mellitus was observed.MAIN OUTCOME MEASURES: Expressions of GFAP mRNA and protein RESULTS: ① Immunohistochemical analysis showed that: When rats suffered from diabetes mellitus for 1 month, taurine can inhibit the immunoreactivity of GFAP. With the elongation of intervention time of taurine, the immunoreactivity of GFAP is weakened. ②RT-PCR detection: Starting from the beginning of 2 months, the expression of GFAPmRNA in diabetic group began to increase; taurine obviously down-regulated the expression of GFAP mRNA. ③ In taurine-treated diabetic group of 3 months,the expression of GFAP was the strongest. When diabetic rats were treated by taurine for 2 months, the expression of GFAP was decreased.CONCLUSION: Taurine can down-regulate the expression of GFAP mR-NA and protein, indicating that it can protect retinal pathological change in rats with diabetes mellitus.

AIM To research the antiasthmatic effects of Zuogui Pills,Yougui Pills and Yupingfeng Powder and their mechanism of action.METHODS Male Brown Norway rats were randomly divided into five groups.OVA was used to induce the model of chronic asthma.Five groups of asthmatic rats were given Zuogui Pills,Yougui Pills,Yupingfeng Powder by gavage once daily for 14 d,respectively.The normal and the model group were given normal saline.Penh value,serum Thl,Th2 inflammatory cytokines,and CROT and ACTH protein levels were determined.Pathological changes of airway remodeling were measured.RT-PCR method was used to measure the expression of CRH mRNA in hypothalamus,and TGF-β1 and Smad3 mRNA in lung tissues.RESULTS Penh values of Yupingfeng Powder group and Yougui Pills group were significantly lower than that of Zuogui Pills group.The Thl inflammatory cytokine of Yupingfeng Powder group was significantly higher than that of Zuogui Pills group,while the Th2 cytokine was significantly lower than that of Zuogui Pills group.In Yougui Pills group,only IL-5 was significantly lower than that of Zuogui Pills group.Serum CROT,ACTH protein and CRH mRNA of Yupingfeng Powder group were significantly higher than those in Zuogui Pills group.No significant difference was found between Yupingfeng Powder group and Zuogui Pills group with regard to TGF-β1 and Smad3 mRNA.Smad3 mRNA of Yougui Pills group was significantly higher than that of the model group.Airway remodeling was significantly reduced by Yupingfeng Powder and Zuogui Pills.The number of goblet cells in Yougui Pills group was significantly lower than that in Zuogui Pills group.CONCLUSION All the three formulaes possess significant antiasthma effects.Regulation effects of Yupingfeng Powder on Thl/Th2 balance,HPA and TGF-β1/Smad3 pathway are strong.Compared with Zuogui Pills,Yougui Pills possesses weaker TGF-β1/Smad3 regulative effects and stronger HPA regulative effects.

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

AIM:To investigate radiosensitization effect of apatinib, a vascular endothelial growth factor (VEGF) receptor2 tyrosine kinase inhibitor, on human gastric carcinoma cell line SGC-7901 and its mechanism.METHODS:SGC-7901 cells were divided into control group, apatinib group, radiotherapy group and combination group.The cell viability was measured by CCK-8 assay.The changes of cell apoptosis and cell cycle were analyzed by flow cytometry.The protein levels of cell apoptosis biomarkers, such as PARP, cleaved caspase-9, cleaved caspase-3 and Bcl-2, and cell proliferation biomarkers, p-PLCγ1 and p-ERK1/2, were detected by Western blot.γ-H2AX expression was detected by immunofluorescence.RESULTS:Compared with apatinib group and radiation group, the cell viability was inhibited after treatment with both apatinib and X-ray (P<0.01).The protein levels of cell proliferation markers p-PLCγ1 and p-ERK1/2 were down-regulated.The cell apoptosis was enhanced (P<0.01).The protein levels of cell apoptosis makers such as PARP, cleaved caspase-9 and cleaved caspase-3 were up-regulated, while Bcl-2 was down-regulated.The disappearance of γ-H2AX foci in the nucleus was delayed, indicating that apatinib impaired the repair of radiation-induced DNA double-strand breaks.The proportion of G2 phase was significantly increased (P<0.01).The combination treatment had more significant effect on SGC-7901 cells than treating with apatinib or radiotherapy alone.CONCLUSION:Apatinib increases the radiosensitivity of gastric cancer cells via blocking VEGF pathway.

Objective To Compare characteristics between the psychological aids hotline callers who had single attempted suicide and those callers who had repeated attempted suicide. Methods The analysis was conducted on the da?tabase of Beijing psychological aids hotline from Dec., 2002 to Dec., 2008. All the effective callers were consulted by pro?fessional hotline operators. During the calling, the callers were interviewed on their attempted suicides, depression and 11 correlates associated with caller's suicidal behaviors. Results Among 4519 callers who had attempted suicide, 2441 (54%) of them had attempted suicide once, and the other 2078 (46.0%) callers had repeated suicide attempts. After ad?justed for gender, age, and the other 4 demographic variables, history of being abused (OR=1.35, 95%CI: 1.08~1.67), afraid of being attack (OR=1.35, 95%CI: 1.01~1.59), relatives or associates with prior suicidal behavior history (OR=1.17, 95%CI: 1.001~1.36), hopelessness (OR=1.20, 95%CI: 1.02~1.40), and psychological treatment history (OR=0.73, 95%CI:0.62~0.86) were associated with repeated suicide among hotline callers. Conclusions To improve the effective?ness of hotline based suicide prevention, assessment of risk of repeated attempted suicide among the callers with a histo?ry of prior attempted suicide should focus on the history of being abused, afraid of being attack, hopelessness, relatives or associates with prior suicide history, and psychological treatment history.

Objective To study the differences in intestinal flora of normal and type 2 diabetic mice, the effect of alcoholic extract of Coreopsis tinctoria Nutt on mouse intestinal flora, and explore the possible relationship between alcoholic extract of Coreopsis tinctoria Nutt, intestinal flora and type 2 diabetes mellitus (T2DM) in mice.Methods Stool samples were collected from the normal control group (A), high dose (1.8 g/kg) (B) and moderate dose (1.2 g/kg) (C) alco-holic extract of Coreopsis tinctoria Nutt model groups, metformin (0.2 g/kg) treatment group (D) and blank control (E) group.16S rDNA real-time quantitative PCR assay was used to determine the levels of Clostridium coccoides and Bacteroides thetaiotaomicron in the stool samples.Pearson analysis method was used to analyze the correlation between the levels of tar-get bacterial species and the fasting blood glucose ( FBG) in the mice.Results 1.Compared with the normal control group, the levels of Clostridium coccoides and Bacteroides thetaiotaomicron in the T2DM model group were significantly low-ered (P=0.017, P=0.002).2.Compared with the model group, the levels of Clostridium coccoides and Bacteroides the-taiotaomicron of the high dose Coreopsis tinctoria Nutt alcohol extract group were significantly different ( 2 weeks: P =0.027, P=0.006;4 weeks:P=0.007, P=0.012).3.The levels of Clostridium coccoides and Bacteroides thetaiotaomi-cron were positively correlated with the FBG level in the mice.Conclusions The alcohol extract of Coreopsis tinctoria Nutt has certain effect on the intestinal flora in type 2 diabetic mice and there is certain correlation between the effect of alcohol extract of Coreopsis tinctoria Nutt and their blood glucose level.

Objective:To investigate the effects of intravascular hypothermia combined with early post-pyloric feeding on the neurological function and prognosis in patients with severe ischemic stroke, and to provide a theoretical basis for clinical decision-making on the optimal nutritional support strategy for patients with severe ischemic stroke during intravascular hypothermia treatment.Methods:This was a retrospective, non-randomized, controlled study. A total of 78 patients with first severe ischemic stroke who were admitted to the ICU of Neurology Department, Xuanwu Hospital, Capital Medical University from January 2018 to December 2021 were selected. General information and clinical data of the patients were collected and grouped according to intrvascular hypothermia combined with nutritional support. Patients were divided into early post-pyloric feeding group of 52 cases and early parenteral nutrition group of 26 cases. The neurological prognosis, disease prognosis, nutritional status and complications related to nutritional support of the two groups were retrospectively analyzed.Results:The Glasgow score at 30th day after intravascular hypothermia in the early postpyloric feeding group was (11.25 ± 4.92) points, which was higher than that in the early parenteral nutrition group (8.40 ± 5.53), and the difference was statistically significant ( t=-2.45, P<0.05). After treatment, the serum total protein and hemoglobin of early postpyloric feeding group were (59.56 ± 5.09) g/L and (131.06 ± 19.58) g/L, respectively, which were higher than those of early parenteral nutrition group (56.52 ± 7.94) g/L and (122.07 ± 17.72) g/L. The difference was statistically significant ( t=-2.03, -1.91, P<0.05). The clinical pulmonary infection score of the early postpyloric feeding group was (7.33 ± 0.96) points, which was lower than that of the early parenteral nutrition group (9.42 ± 2.11). The mechanical ventilation time and ICU stay time were (17.46 ± 10.47) days and (28.89 ± 12.59) days, respectively. Compared with the early parenteral nutrition group (25.77 ± 15.20) days and (37.07 ± 17.15) days, the differences were statistically significant ( t=3.28, 2.83, 2.52, all P<0.05). There were no significant differences in catheter-associated bloodstream infection and ICU hospitalization mortality between the two groups (both P>0.05). Conclusions:Intravascular hypothermia combined with early post-pyloric feeding can improve the nutritional status of patients with severe ischemic stroke, effectively control pulmonary infection, shorten mechanical ventilation and hospital stay, and promote neurological repair.

Objective The study aims to optimize the conditions for constructing orthotopic nude mouse models of liver cancer by injecting human liver tumor cell lines and to explore appropriate timings for drug administration. Methods Human hepatocellular carcinoma Hep3B and hepatoblastoma HepG2 cell lines, which stably expressing the luciferase reporter gene (LUC), were selected. The linear correlation between the luciferase luminescence intensity and the number of liver tumor cells was analyzed using a Small Animal In Vivo Imaging system to verify the luminescent efficiency of the human liver tumor cells. Different concentrations (8×10⁶, 2.4×10⁷, 7.2×10⁷ cells/mL) and resuspension media (PBS, Matrigel) of human liver tumor cell suspensions HepG2-LUC and Hep3B-LUC were orthotopically inoculated into the liver lobes of 5-week-old female BALB/c nude mice (12 groups, 7 mice each) to construct human liver tumor nude mouse orthotopic cancer models. Every 7 days, the weights of mice were recorded, and the growth of orthotopic tumors was monitored using the Small Animal In Vivo Imaging system. On day 35 post-cell inoculation, mouse livers were dissected, and pathological slices were prepared for HE staining to observe histopathological changes in liver tissues. Results The luminescence intensity of human liver tumor cell lines was positively correlated with the number of cells (R²=0.983 1, R²=0.970 5), indicating their suitability for orthotopic model construction. Successful modeling was achieved in the high-concentration groups of HepG2-LUC, the low-, medium-, and high-concentration groups of HepG2-LUC+Matrigel, the medium- and high-concentration groups of Hep3B-LUC, and the low-, medium-, and high-concentration groups of Hep3B-LUC+Matrigel. For both HepG2-LUC+Matrigel and Hep3B-LUC+Matrigel groups, mice in the high-concentration groups exhibited significantly reduced body weight compared to the low- and medium-concentration groups (both with P<0.05). The luminescence intensity of successfully modeled mice increased exponentially over time (R²>0.950 0), and reached a minimum of 1.0×10⁷ p/(s·cm²·sr) by day 14 post-transplantation. Mice in the low- and medium-concentration groups of HepG2-LUC and the low-concentration group of Hep3B-LUC showed no significant pathological changes, while the other groups exhibited evident liver tumors and hepatocyte lesions. Conclusion For the HepG2-LUC cell line, the recommended injection volume is 50 µL with a cell density of 2.4×10⁷ cells/mL, resuspended with Matrigel, followed by drug administration or prognostic measures on day 7 post-modeling. For the Hep3B-LUC cell line, the recommended injection volume is 50 µL with a cell density of 7.2×10⁷ cells/mL, not resuspended with Matrigel, with administration or prognostic measures on day 14 post-modeling.

ObjectiveTo observe the clinical effect of modified Yinqiaosan combined with antibiotics in the treatment of acute tonsillitis in children with wind-heat invading lung syndrome. MethodA total of 96 children with acute tonsillitis of wind-heat invading the lung syndrome were randomized into control group and observation group. The control group was treated with routine antibiotics， and the observation group was treated with modified Yinqiaosan and antibiotics for 7 days. The scores of major symptoms （sore throat， erythmatous throat， dysphagia， erythmatous tonsils and suppuration） and minor symptoms （fever， cough， stool， and tongue） and the levels of inflammation- and immune-related indexes ［white blood cell （WBC）， C-reactive protein （CRP）， serum amyloid A （SAA）， interleukin-6 （IL-6）， and interferon-γ （IFN-γ）］ were compared between two groups. ResultThe data of 92 children were statistically analyzed： 45 in the observation group and 47 cases in the control group. The total clinical effective rate of the observation group was 95.56%， as compared with the 93.62% of the control group. After treatment， the scores of major symptoms in the observation group were lower than those in the control group （P<0.05）， and the scores of cough， defecation， and tongue in the observation group were lower than those in the control group （P<0.05）. The levels of inflammation- and immune-related indexes after treatment in the observation group were lower than those before treatment （P<0.05）. Except IFN-γ， the levels of the inflammation- and immune-related indexes in the control group were lower than those before treatment （P<0.05）. After treatment， the levels of SAA and IL-6 in the observation group were lower than those in the control group （P<0.05）. ConclusionModified Yinqiaosan combined with antibiotics can significantly reduce the expression of SAA and IL-6 in the treatment of children with acute tonsillitis， thereby alleviating inflammation and clinical symptoms and improving prognosis.

ObjectiveTo explore effect of modified Wuhutang on airway inflammation and expression of mucin （Muc） 5AC， signal transducer and activator of transcription 3 （STAT3）， nuclear factor kappa B （NF-κB）， and NOD-like receptor thermal protein domain associated protein 3 （NLRP3） in respiratory syncytial virus （RSV）-infected asthmatic mice. MethodSeventy male BALB/c mice of 6-8 weeks old were randomized into normal control （CON）， asthma （ovalbumin， OVA）， RSV infection-induced asthma （OVA+RSV）， high-， medium-， and low-dose （4.08， 2.04， 1.02 g·kg^-1·d^-1， respectively） modified Wuhutang， and dexamethasone （Dxms， 0.1 g·kg^-1d^-1） groups （n=10）. The model of asthma was established by sensitization and atomization inhalation with OVA. The RSV infection-induced asthma model was established by three consecutive RSV nasal infusions （1.0 × 10⁶ PFU·mL^-1， 50 μL）. Wuhutang was administrated by gavage， and Dxms by intraperitoneal injection. The CON group was given the same amount of normal saline by gavage. The mice were anesthetized with 2.5% pentobarbital sodium 24 h after the last administration， and then the lung tissue was stained by hematoxylin-eosin （HE） and Van Gieson （VG） for observation of airway inflammation. The immunohistochemical assay was employed to detect the expression of Muc5AC. Western blot was employed to determine the protein levels of phosphorylated （p）-STAT3， STAT3， p-NF-κB， NF-κB， and NLRP3. ResultCompared with the CON group， the OVA group presented airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition. The OVA+RSV group showed severer airway inflammatory cell infiltration and tissue hyperemia and edema than the OVA group. Compared with the OVA+RSV group， modified Wuhutang alleviated the airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition， and the high-dose group had the best performance. Compared with the CON group， the OVA group and the OVA+RSV group showed increased expression level of Muc5AC （P<0.01）. Compared with the OVA+RSV group， modified Wuhutang reduced the expression level of Muc5AC， and the reduction was significant in the high-dose group （P<0.05）. Compared with the high-dose modified Wuhutang group， Dxms lowered the expression level of Muc5AC （P<0.05）. Compared with the CON group， the OVA and OVA+RSV groups showed up-regulated protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.05， P<0.01）. Compared with the OVA+RSV group， modified Wuhutang down-regulated the protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.01）. Compared with the high-dose modified Wuhutang group， the Dxms group showed up-regulated levels of p-STAT3， p-NF-κB proteins （P<0.01）. ConclusionModified Wuhutang can reduce airway inflammation and down-regulate the expression of Muc5AC， p-STAT3， p-NF-κB， and NLRP3 in RSV-infected asthmatic mice， which suggests that Wuhutang reduces airway inflammation in RSV-infected asthma by regulating the STAT3/NF-κB signaling pathway.