Objective:To optimize the ethanol precipitation technique for Fufang Shenqi soft capsules. Methods: An orthogonal design was used to optimize the technique with the relative density of the concentrated solution, ethanol concentration, standing time, temperature of ethanol precipitation as the influencing factors and the yield of dry extract and the content of total polysaccharides as the indices. Results:The best ethanol precipitation technique was as follows:the relative density of the concentrated solution was 1. 10, 95% ethanol was used to obtain 60% ethanol concentration, and the standing time was 48 h under the temperature of 10-30℃. Con-clusion:The optimized ethanol precipitation technique for Fufang Shenqi soft capsules is simple and practicable, and suitable for prac-tical production.

Objective To screen the serum protein molecular markers of postmenopausal osteoporosis by the proteomicsanalysis using Tandem Mass Tag (TMT) combined with liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS).Methods 20 serum protein samples were recruited (10 cases of postmenopausal patients with osteoporosis and 10 cases of postmenopausal women without osteoporosis)and the high abundance ratios protein was removed,differentiation protein was extracted and labeled with TMT reagent.Then,mass spectrometric detection,data analysis of differentially expressed proteins,and analysis of biological information were carried out.Results 87 significantly differentially expressed proteins were screened from the differentiated protein expression profile by LC-ESI-MS/MS combined with TMT labeling.While 50 proteins were up-regulated,and 37 proteins were down-regulated.Differentially expressed proteins were analyzed by GO annotation,these proteins are mainly involved in 15 kinds of biological processes,7 kinds of cellular component,6 kinds of molecular function.RAB7A,TSP1,GAS6,SPP24 were screenedas candidate proteins which were related to mechanism of bone remodeling of osteoporosis.By STRING 10.0 protein interaction network analysis tools,RAB7A,TSP1,GAS6 were located in the center of the interaction network.SPP24 was located at edge of the network,but it is directly related to the protein BMP-2 of bone remodeling.RAB7A,TSP1,GAS6,SPP24 may be associated with the pathogenesis of postmenopausal osteoporosis.Conclusion These results provide that the proteomics analysis by using TMT coupled with LC-ES1-MS/MS was a feasible method for screening the molecular biomarkers.It suggests that RAB7A,TSP1,GAS6 and SPP24 may be useful biomarkers which can be used both in diagnosis and treatment of postmenopausal osteoporosis.

Objective: To explore the effect of the new vaginal douche device on vaginal irrigation before ovulation extraction in vitro fertilization-embryo transfer. Methods: From August 2017 to April 2018, 100 cases of infertility patients admitted to the first hospital of reproductive and gynecology of Jilin University who had to undergo ovulectomy were selected and divided into 50 cases in the experimental group and 50 cases in the control group according to the random number table. In the test group, a new type of vaginal irrigator was used for vaginal irrigation one day before the operation, while in the control group, a disposable enema was used for vaginal irrigation. The vaginal cleanliness and comfort of the two groups were compared after washing. Results: Vaginal cleanliness was compared between the two groups. Bacterial culture of vaginal secretions was positive in 2 patients in the experimental group, with a positive rate of 4%. In the control group, bacterial culture of vaginal secretions was positive in 14 patients, with a positive rate of 28%. Vaginal cleanliness was better in the experimental group than in the control group (χ²=10.714, P=0.001). In the experimental group, there were 2 cases of discomfort, 10 cases of discomfort, 29 cases of comfort, and 9 cases of comfort. In the control group, there were 8 cases of discomfort and 17 cases of discomfort. The comfort level of the experimental group was higher than that of the control group, and the difference was statistically significant (Z=-2.271, P=0.023). Conclusions The operation method of the new vaginal douche is simple and soft, which can not only effectively and thoroughly clean the vagina, reduce the incidence of infection, but also increase the comfort level of patients, alleviate psychological fear and improve patient satisfaction.

Objective The aim of this study was to investigate the polymorphism of SLA class II genes in Canadian SPF Yorkshire and Landrace pigs.Methods Blood samples were obtained from 15 SPF Yorkshire and 22 Landrace pigs for isolation of peripheral blood mononuclear cells respectively, and the DQB1, DRB1 and DQA genes were amplified by PCR after reverse transcription.SLA class II genes were obtained by analyzing the direct and cloning result.The polymorphism of alleles was analyzed using the DNAsp 5.0 software.Results A total of 25 alleles were identified at three genes, including eight DQB1, ten DRB1 and seven DQA, and three alleles were submitted the complete sequences for the first time.The official allele names were assigned as SLA-DQB1*0212 (KU754590), SLA-DQB1*0203 (KU754591) and DRB1*06:07(KU754601) by the SLA Nomenclature Committee.Three novel DQA alleles were discovered.Five of the 15 amino acids, one of the 16 amino acids and 11 of the 19 amino acids, which bind processing antigens, showed well conserved among the alleles of DQB1, DRB1 and DQA genes in the SPF Yorkshire and Landrace pigs, respectively.Neighbor-joining tree showed that the three genes were divided into two clusters, respectively.There was a close relationship between SPF Yorkshire and Landrace pigs and foreign Yucatan miniature pigs, and it showed no obvious genetic distance with other pigs.Conclusions A total of 25 SLA class II alleles have been identified successfully in this study, and there are more abundant polymorphism for them.There is a widely distribution for SLA class II alleles identified in this study in other pig breeds.It is critical for the eventual future use of SPF Yorkshire and Landrace pigs as classical laboratory animal models.

ObjectiveTo study the effect of low molecular weight heparin sodium (LMWHS) therapy for exertional heat stroke (EHS) patients with pre-disseminated intravascular coagulation (pre-DIC).Methods A prospective randomized controlled trial (RCT) was conducted. Thirty-six patients with EHS with pre-DIC admitted to Department of Critical Care Medicine of 180th Hospital of Chinese PLA from April 2012 to November 2014 were divided into heparin sodium group (n = 20) and LMWHS group (n = 16) in accordance with the random number table. All patients received bundle treatment after being admitted to the hospital, including rapid cooling, fluid resuscitation, organ support (mechanical ventilation, hemopurification if necessary), supplement of pro-coagulation factors, etc. The patients in heparin sodium group were treated with continuous heparin sodium 12 500 U throughout 24 hours with intravenous pump for 5 days, and the patients in LMWHS group were given LMWHS 2 500 U subcutaneously, twice a day for 5 days.The incidence of DIC, incidence of bleeding and mortality of two groups were compared.The platelet count (PLT), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib) and D-dimer of each patient between pre and post treatment times were compared.Results No significant difference was found in the incidence of DIC and mortality between LMWHS group and heparin sodium group (31.2% vs. 30.0%,χ2 =0.007,P = 0.936; 6.2% vs. 5.0%,χ2 = 0.026,P = 0.871). Incidence of bleeding during treatment in LMWHS group was significantly lower than that in heparin sodium group (12.5% vs. 45.0%,χ2 = 4.425,P = 0.035). After treatment,PLT in both LMWHS group and heparin sodium group was significantly increased compared with that before treatment (×109/L: 140.5±17.5 vs. 110.5±16.5, 152.6±21.5 vs. 120.0±20.0, bothP< 0.05) and D-dimer was significantly decreased (mg/L: 0.5±0.1 vs. 3.2±1.2, 0.6±0.2 vs. 4.4±1.8, bothP< 0.05). APTT after treatment in heparin sodium group was significantly prolonged compared with that before treatment (s: 75.3±10.6 vs. 44.1±8.2,P< 0.05) while no change in APTT was found in LMWHS group (s: 38.6±5.5 vs. 42.1±8.4,P> 0.05). No significant difference was found in PT and Fib between pre and post treatment in all the patients.Conclusion When LMWHS was applied in EHS patients in pre-DIC stage, it could not only prevent DIC as efficiently as heparin sodium, but also results in lower incidence of bleeding. So LMWHS is safer.

Objective To investigate the changes of the expression of HIF-1α (hypoxia induciblefactor-1a) mRNA in myocardium of rats after cardiopulmonary resuscitation (CPR) and the intervention effects of exogenous hydrogen sulfide on it. Method Forty five male SD rats were randomly divided into three groups: control group ( n = 15 ), cardiopulmonary resuscitation group ( CPR group, n = 15 ) and NaHS + CPR group (n = 15 ) . Rats of control group were anesthetized and intubated without asphyxia and cardiac arrest. The rats of CPR group and NaHS + CPR group were operated to induce cardiac arrest by asphyxiation. In the rats of NaHS + CPR group, NaHS in dose of 50 ug/kg was administrated via the femoral venous line 1 minute before CPR. Hemodynamic was monitored continuously. The expression of HIF - lα mRNA in myocardium of rats in each group was determined by using RT-PCR and the activity of myeloperoxidase (MPO) in the myocardium of rats in each group was assayed by using a patent reagent box 6 h after CPR. The histopathological changes of myocardium were also observed. The t- test was used for statistical analysis. Results There was no statistically significant difference in hemodynamic changes between CPR group and CPR + NaHS group ( P ＞ 0. 05 ) . When compared with the control group, the activity of MPO and the expression of HIF-1α mRNA in CPR group and CPR + NaHS group were both increased, and those increased in CPR + NaHS group was more significant (P ＜ 0. 05) . When compared with CPR group, the expression of HIF-1α mRNA in CPR + NaHS group was higher, however, the activity of MPO in CPR + NaHS group was lower ( P ＜ 0. 05) . There were various histopathological changes of myocardium of rats found in CPR group and CPR + NaHS group, and the damage of myocardium of rats in CPR group was more obvious than that in CPR + NaHS group. Conclusions The expression of HIF-1α mRNA in myocardium of rats was increased after CPR. Exogenous hydrogen sulfide can protect myocardium cells from resuscitation-perfusion injury, and the protection is associated with increase in expression of HIF 1 αmRNA.

Objective To investigate the effects of hydrogen sulfide on sepsis-induced cardiac dysfunction in rats.Methods Male SD rats were randomly ( random number) divided into 4 groups:control group ( Ⅰ group),sepsis group ( Ⅱ group),sepsis + NaHS group (Ⅲ group),sepsis + PAG group (Ⅳ group). Cecal ligation and puncture technique was used in Ⅱ, Ⅲ, Ⅳ group. Hemodynamic was observed,and H2S synthesis,CSE (cystathionine-r-lyase) mRNA,MPO activity and the level of TNF-α,IL-1β were determined.The morphological changes and infiltration of inflammatory cells in myocardium were also observed.Results Compared with Ⅰ group,H2S synthesis,the levels of TNF-α,IL-1 β,the activity of MPO increased ( P ＜ 0.05 or P ＜ 0.01 ),and the expression of CSE-mRNA increased,and blood pressure of rats decreased significantly in Ⅱ group.Compared with Ⅱ group,H2S synthesis,the levels of TNF-α,IL-1β and the activity of MPO increased (P ＜ 0.05),the expression of CSE mRNA did not change noticeably ( P ＞ 0.05),and blood pressure of rats decreased more significantly in Ⅲ group.Compared with Ⅱ group, H2S level decreased,the levels of TNF-α,IL-1β and the activity of MPO decreased significantly,the expression of CSE mRNA decreased and blood pressure of rats decreased in Ⅵ group (P ＜0.05).Histopathological changes of myocardium were aggravated in the following severity order: Ⅰ ＜ Ⅳ ＜＜ Ⅲ.Conclusions CSE/H2S system of the myocardium was upregulated in sepsis rats.Hydrogen sulfide could raise the levels of MPO,TNF-α,IL-1β,aggravating myocardial injury. Contrarily,the inhibitor of H2S could counteract it.

To investigate the interaction and involvement of sodium hydrosulfide (NaHS), a H(2)S donor, on hippocampus of rats suffering from sepsis-associated encephalopathy, rats were subjected to cecal ligation and puncture (CLP)-induced sepsis. Adult male Sprague-Dawley rats were randomly divided into four groups: Sham group, CLP group, CLP+NaHS group and CLP+aminooxyacetic acid (AOAA, an inhibitor of H(2)S formation) group. The four groups were observed at 3, 6, 9, 12 h after treatment. We examined hippocampal H(2)S synthesis and the expression of cystathionine-β-synthetase (CBS), a major enzyme involved in the H(2)S synthesis in hippocampus. CBS expression was detected by reverse transcription polymerase chain reaction (RT-PCR). The concentrations of inflammatory cytokines (TNF-α, IL-1β) were determined in hippocampus by using enzyme-linked immunosorbent assay (ELISA). Neuronal damage was studied by histological examination of hippocampus. In CLP group, H(2)S synthesis was significantly increased in hippocampus compared with sham group and it peaked 3 h after CLP (P<0.05). Sepsis also resulted in a significantly upregulated CBS mRNA in hippocampus. The levels of TNF-α and IL-1β in the hippocampus were substantially elevated at each time point of measurement (P<0.05), and they also reached a peak value at about 3 h. Administration of NaHS significantly aggravated sepsis-associated hippocampus inflammation, as evidenced by TNF-α and IL-1β activity and histological changes in hippocampus. In septic rats pretreated with AOAA, sepsis-associated hippocampus inflammation was reduced. It is concluded that the rats subjected to sepsis may suffer from brain injury and elevated pro-inflammatory cytokines are responsible for the process. Furthermore, administration of H(2)S can increase injurious effects and treatment with AOAA can protect the brain from injury.

To investigate the interaction and involvement of sodium hydrosulfide (NaHS),a H2S donor,on hippocampus of rats suffering from sepsis-associated encephalopathy,rats were subjected to cecal ligation and puncture (CLP)-induced sepsis.Adult male Sprague-Dawley rats were randomly divided into four groups:Sham group,CLP group,CLP+NaHS group and CLP+aminooxyacetic acid (AOAA,an inhibitor of H2S formation) group.The four groups were observed at 3,6,9,12 h after treatment.We examined hippocampal H2S synthesis and the expression of cystathionine-β-synthetase (CBS),a major enzyme involved in the H2S synthesis in hippocampus.CBS expression was detected by reverse transcription polymerase chain reaction (RT-PCR).The concentrations of inflammatory cytokines (TNF-α,IL-1β) were determined in hippocampus by using enzyme-linked immunosorbent assay (ELISA).Neuronal damage was studied by histological examination of hippocampus.In CLP group,H2S synthesis was significantly increased in hippocampus compared with sham group and it peaked 3 h after CLP (P＜0.05).Sepsis also resulted in a significantly upregulated CBS mRNA in hippocampus.The levels of TNF-α and IL-1β in the hippocampus were substantially elevated at each time point of measurement (P＜0.05),and they also reached a peak value at about 3 h.Administration of NaHS significantly aggravated sepsis-associated hippocampus inflammation,as evidenced by TNF-α and IL-1β activity and histological changes in hippocampus.In septic rats pretreated with AOAA,sepsis-associated hippocampus inflammation was reduced.It is concluded that the rats subjected to sepsis may suffer from brain injury and elevated pro-inflammatory cytokines are responsible for the process.Furthermore,administration of H2S can increase injurious effects and treatment with AOAA can protect the brain from injury.

Objective Grey red-backed voles (Myodes rufocanus) are agile, fierce and hard to catch, thus, it is difficult to judge their gender by external appearance, especially for the juvenile voles. Therefore, it may cause difficulties to their allocation and later breeding in laboratories. The aim of this paper is to establish a rapid, simple and accurate method for gender identification of grey red-backed voles. Methods Fresh hair follicles were taken from 6 adult male voles, 3 adult females and 14 4-week-old juvenile voles, 5 male and 5 female 9-week-old Wistar rats, and 5 male and 3 female 6-week-old BALB/c mice. The genomic DNA was extracted using Chelex-100 resin and the zinc-finger Y/X gene (ZFY/ZFX) and the gene of sex-determining region of the Y (SRY) chromosome were amplified by PCR, and a double PCR amplification method was established. Results The ZFY/ZFX gene and SRY gene were simultaneously amplified from the male voles, while only the ZFY/ZFX gene was amplified from the females. The gender of all 23 voles, 10 Wistar rats and 8 BALB/c mice were correctly identified with this method, and the PCR results were consistent with the phenotypic and autopsy results. Conclusions Using fresh hair follicles as experimental materials for gender identification of grey redbacked voles can alleviate shock and damage to the animals. The established double PCR amplification method is accurate, simple, rapid, and deserves to be used for gender identification of grey red-backed voles.