OBJECTIVETo observe the effect of Xinfeng Capsule (XFC) on ankylosing spondylitis (AS) patients' symptoms and signs, serum immunoglobulin levels, peripheral blood lymphocyte autophagy protein, autophagy gene, and to explore its mechanism.

METHODSTotally 59 AS patients were assigned to the treatment group (39 cases) and the control group (20 cases) according to random digit table. Patients in the treatment group received XFC, 0.5 g each pill, three pills each time, 3 times per day, while those in the control group received sulfasalazine (SASP), 0.25 g per tablet, 4 tablets each time, twice per day. Three months consisted of one therapeutic course. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) were statistically calculated. Serum immunoglobulins (IgG1, IgG2, IgG3, IgG4, IgA , SIgA, and IgM) were detected using ELISA. Changes of Beclin1, LC3-II, phosphatidylinositol 3-kinase (PI3K), Akt, the mammalian target of rapamycin (mTOR) were detected using Western blot. Serum autophagy related genes such as Atg1, Atg5, Atg12, Atg13, and Atg17 were detected using the polymerase chain reaction (PCR). The correlation between immunoglobulin subtypes and autophagy gene in AS patients using Spearman correlation.

RESULTSCompared with before treatment, BASDAI, IgG1, lgG3, and IgA decreased (P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.01); ATG1, ATG12, ATG13, and ATG17 mRNA expressions decreased, ATG5 mRNA expression increased (P < 0.01) in the treatment group. But BASDAI, IgG1, and IgA levels decreased (P < 0.05, P < 0.01); PI3K, Akt, and mTOR protein expressions decreased (P < 0.05); ATG1 and ATG13 mRNA expressions decreased (P < 0.05, P < 0.01) in the control group. Compared with the control group, BASDAI, IgG1, and IgA levels decreased (P < 0.05); PI3K, Akt, mTOR protein expressions decreased (P < 0.01); ATG12 and ATG17 mRNA expression decreased, ATG5 mRNA expression increased (P < 0.01) in the XFC group. Correlation analysis showed AS patients' IgG1, IgG2, IgG3, IgA, SIgA, IgM had negative correlation with ATG17; IgG4 and ATG17 were positively correlated (P < 0.05, P < 0.01).

CONCLUSIONXFC could elevate clinical efficacy of AS patients and enhance their autophagy, which might be achieved by acting on PI3K/Akt/mTOR signal, affecting autophagy gene and autophagy protein expression, taking part in the regulation of proliferation and differentiation of lymphocyte B, and strengthen humoral immunity.

Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; Beclin-1 ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Lymphocytes ; drug effects ; Membrane Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Spondylitis, Ankylosing ; drug therapy ; Sulfasalazine ; therapeutic use ; TOR Serine-Threonine Kinases ; metabolism

Objective To explore the effect of "peer mentor" teaching mode on nursing students'psychological stress in early clinical practice (three months).Methods One hundred and sixty nursing students were randomly divided into the study group and the control group,each with 80 cases.The study group received "peer mentor" teaching mode,and the control group received convention teaching mode.The Symptom Checklist-90 (SCL-90),Self-rating Anxiety Scale (SAS),Self-rating Depression Scale (SDS) were used to evaluate the students' psychological state.The clinical comprehensive skill was assessed after 12 weeks.Results The cases of normal,mild anxiety,moderate anxiety,severe anxiety according to SAS were respectively 51,20,8,1 in the study group,and 33,30,13,4 in the control group respectively,and the difference was statistically significant (x2 =8.275,P ＜ 0.01).The cases of normal,mild depression,severe depression,moderate depression numbers according to SDS were respectively 51,20,8,1 in the study group,and 34,31,12,3 in the control group respectively,and the difference was statistically significant (x2 =4.971,P ＜ 0.05).The differences were statistically significant in the interpersonal sensitivity (x2 =5.71,P ＜ 0.05),hostile (x2 =5.13,P ＜ 0.05)and paranoid (x2 =4.78,P ＜ 0.05) between the two groups.In the clinical practice,cooperation consciousness,love consciousness,psychological ability,interpersonal communication ability,health education ability and the evaluation feedback in the study were better than those of the control group,and the differences were statistically significant (x2 =9.49,10.25,11.62,6.32,8.07,7.17,respectively; P ＜ 0.05).Conclusions Peer mentor teaching mode can reduce students' psychological stress in early clinical practice,and help to improve practice quality.

Objective To evaluate the antibody titer distributions after primary vaccination by different sequential schedules of Sabin strain-based inactivated poliovirus vaccine(sIPV) and bivalent oral attenuated live poliomyelitis vaccine against types 1 and 3 (bOPV) in Drug Candy(DC) form or liquid dosage form. Methods Eligible infants of 2 months old selected in Liuzhou were assigned randomly in a ratio of 1:1:1:1 to 4 groups as following: sIPV+2bOPV(DC), sIPV+2bOPV(liquid), 2sIPV+bOPV(DC), 2sIPV+bOPV(liquid), and were vaccinated at 0, 28, 56 days. Polio neutralizing antibody titers against poliovirus types 1, 2 and 3 were tested prior to Dose 1 and at 28 days after Dose 3. Results The antibody titer distribution for type 1 was statistically different between sIPV+2bOPV(DC) and sIPV+2bOPV(liquid) (Z=-2.589, P=0.010) while no significant differences were detected between the two groups for type 2(Z=-0.331, P=0.741) and type 3(Z=-1.556, P=0.120). There were no significant differences between 2sIPV +bOPV(DC) and 2sIPV+bOPV(liquid) for the distributions(All P>0.05) (type 1: Z=-1.249, P=0.212; type 2: Z=-1.658, P=0.097; type 3: Z=-1.436, P=0.151). In the same dosage forms with different sequential schedules, the antibody titer distributions were significantly different between 2 doses sIPV and 1 dose sIPV groups(All P<0.05)(sIPV+2bOPV(liquid) vs 2sIPV+bOPV(liquid): type 1: Z=-2.766, P=0.006; type 2: Z=-9.137, P<0.001; type 3: Z=-5.529, P<0.001. sIPV+2bOPV(DC) vs 2sIPV+bOPV(DC): type 1: Z=-3.748, P<0.001; type 2: Z=-7.660, P<0.001; type 3: Z=-6.030, P<0.001). Conclusions Different dosage forms have similar immune effects, so appropriate dosage forms should be selected for vaccination according to the effectiveness, characteristics of subjects and the population density. In the case of sufficient supply of sIPV, 2 doses sIPV sequential program should be the first choice to complete the primary immunization.

Objective To study the chemical constituents from Bletilla ochracea. Methods The compounds were extracted by 95％ alcohol and isolated by column chromatography on silica gel and Sephadex LH-20. Their structures were identified by spectroscopic analysis (～1 H NMR and 13 CNMR).Results Nine compouds were obtained and identified as lusianthridin (1), 1, 2, 7-trihydroxy-4-methoxy-9, 10-dihydroxyphenanthrene (2), nudol (3), coelonin (4), batatasin Ⅲ (5), 3, 7-dihydroxy-2, 4-dimethoxyphenanthrene (6), daucosterol (7), β-sitosterol (8), stigmasterol (9).Conclus ion Compounds 2, 3, 5 were isolated from this plant for the first time.

Objective: To explore GRACE (global registry of acute coronary events)score on short term prognosis of ST-segment elevation myocardial infarction (STEMI)in patients elder than 75 years with primary percutaneous coronary intervention(PCI). Methods: A total of 104 STEMI patients elder than 75 years with primary PCI in our hospital from 2011-11 to 2014-01 were studied. Based on GRACEscore at admission, the patients were divided into 2 groups: Lower/mid risk group, n=72 patients with GRACEscore at 112-154 (136.5±10.6) and High risk group, n=32 patients with GRACE score at 155-202(167.8±12.3). The baseline condition and outcomes were compared between 2 groups and the primary endpoint was 1 year mortality. Predictive value of GRACEscore on 1 year mortality was evaluated by ROC curve, the relationships between Lower/mid risk group, High risk group and clinical outcomes were assessed by log-ranksurvive curve andunivariate Cox regression analysis. Results: The area under ROC curve for GRACEscore predicting 1 year mortality was 0.788 with the sensitivity at 70.0%and specificity at 84.0 %.Univariate Cox regression analysis indicated that compared with Lower/mid risk group, High risk group had the higher risk of 1-year death (HR=5.75, 95% CI 1.486-22.256, P=0.0113); log-rank survive curve presented that High risk group had the higher 1 year mortality (21.9% vs 4.2%, P=0.0039). Conclusion: GRACE score may further distinguish the lower/mid risk and high risk populations in elder STEMI patients; it may also predict 1 year clinical prognosis.

Objective:To investigate the association of cadmium exposure with liver function among adults in a non-ferrous metal mining area in Guangxi.Methods:A total of 310 residents aged 18 and above were recruited from 5 heavy metals polluted villages in a non-ferrous metal mining area in Guangxi from 2013 to 2014. The general demographic characteristics, blood cadmium levels and indicators of liver function index [Total bilirubin (TBIL), Glutamic oxaloacetic transaminase (AST), Alanine transaminase (ALT) and Glutamine transaminase (GGT)] were obtained by using questionnaire, physical examination and laboratory test. The blood cadmium levels were divided into quartiles as Q1- Q4 groups (using Q1 group as the reference).Multivariate logistic regression model was used to analyze the correlation between the blood cadmium level and functional liver index. Results:The age of subjects was (49.2±15.4) years, and 112 (36.1%) subjects were male residents. The prevalence of abnormal rates of TBIL,AST,ALT and GGT were 17.4% (54), 19.7% (61), 10.7% (33) and 11.9% (37), respectively. The geometric mean value of cadmium levels in adults was 3.72(95% CI: 3.43-4.02) μg/L. After adjusting for age, gender, body mass index (BMI), smoking, drinking, total cholesterol, hypertriglyceridemia and other factors, the risk of abnormal AST index in the highest concentration of blood cadmium group ( Q4) was higher than that in the lowest concentration of blood cadmium group ( Q1) ( OR=2.92, 95%CI:1.07-7.98). Conclusion:The level of blood cadmium exposure is higher than the reference value of general population in China, and the elevated cadmium exposure is related to the increasing risk of AST abnormality.

Krüppel-like transcription factor 2 (KLF2) plays a key regulatory role in endothelial inflammation, thrombosis, angiogenesis and macrophage inflammation and polarization, and up-regulation of KLF2 expression has the potential to prevent and treatment atherosclerosis. In this study, trichostatin C (TSC) was obtained from the secondary metabolites of rice fermentation of Streptomyces sp. CPCC 203909 as a KLF2 up-regulator by using a high throughput screening model based on a KLF2 promoter luciferase reporter assay. TSC significantly inhibited the adhesion of tumor necrosis factor-α (TNFα) induced monocytes (THP-1) to human umbilical vein endothelial cells (HUVECs). Western blot results showed that TSC decreased TNFα induced the protein expression increase of vascular cell adhesion molecule-1 (VCAM-1), and thereby inhibited endothelial inflammation. The results of histone deacetylase (HDAC) overexpression and molecular docking experiments showed that TSC upregulated the expression of KLF2 by inhibiting subtypes of HDAC 4/5/7. In conclusion, this study suggests that TSC up-regulates the expression of KLF2 through inhibiting HDAC 4/5/7 and thus inhibits TNFα induced endothelial inflammation, and it has the potential to prevent and treat atherosclerosis.

Objective:To investigate the association of cadmium exposure with liver function among adults in a non-ferrous metal mining area in Guangxi.Methods:A total of 310 residents aged 18 and above were recruited from 5 heavy metals polluted villages in a non-ferrous metal mining area in Guangxi from 2013 to 2014. The general demographic characteristics, blood cadmium levels and indicators of liver function index [Total bilirubin (TBIL), Glutamic oxaloacetic transaminase (AST), Alanine transaminase (ALT) and Glutamine transaminase (GGT)] were obtained by using questionnaire, physical examination and laboratory test. The blood cadmium levels were divided into quartiles as Q1- Q4 groups (using Q1 group as the reference).Multivariate logistic regression model was used to analyze the correlation between the blood cadmium level and functional liver index. Results:The age of subjects was (49.2±15.4) years, and 112 (36.1%) subjects were male residents. The prevalence of abnormal rates of TBIL,AST,ALT and GGT were 17.4% (54), 19.7% (61), 10.7% (33) and 11.9% (37), respectively. The geometric mean value of cadmium levels in adults was 3.72(95% CI: 3.43-4.02) μg/L. After adjusting for age, gender, body mass index (BMI), smoking, drinking, total cholesterol, hypertriglyceridemia and other factors, the risk of abnormal AST index in the highest concentration of blood cadmium group ( Q4) was higher than that in the lowest concentration of blood cadmium group ( Q1) ( OR=2.92, 95%CI:1.07-7.98). Conclusion:The level of blood cadmium exposure is higher than the reference value of general population in China, and the elevated cadmium exposure is related to the increasing risk of AST abnormality.

Chemokine signal pathways are important for the regulation of tumour metastasis. Chemokine receptors CXCR4 (C-X-C chemokine receptor type 4) and XCR1 (chemokine XC receptor 1) are involved in the metastasis of breast cancer, while the interaction between them remains unclear. Here we first identified the interaction between CXCR4 and XCR1 based on membrane protein yeast two-hybrid assays. Bioluminescence resonance energy transfer (BRET) showed that XCR1 could competitively bind to CXCR4 to form a heterodimer (P < 0.01). Results of wound healing assays via transient transfection of XCR1 and CXCR4 into HEK293 cells showed that 41.55% of the migration area rate in the co-transformation group was lower than 58.75% in the CXCR4-alone group after adding 30 nmol/L S D F-β. The co-expression of XCR1 inhibited the cellular motility, possibly mediated by the SDF-1β (stromal cell-derived factor 1)/CXCR4 signal pathway (P < 0.05). Furthermore, CXCR4 on the cell surface after co-expression of XCR1 in CXCR4-EGFP transgenic HEK293 cells was detected by flow cytometry. And the result suggested that XCR1 could accelerate the internalization of CXCR4 into the heterodimer induced by 30 nmol/L SDF-1β (P<0.05), which increased the internalization rate from 14.38% to 64.10%. Finally, the phosphorylation of Akt and ERK, which were involved in cell proliferation and migration, respectively, were examined. After 10 minutes of SDF-1β stimulation, ERK phosphorylation in the CXCR4-alone group showed a 3.59-fold increase, whereas the increase of ERK phosphorylation in the co-transfected group was only 2.08-fold. Interestingly, heterodimer formation reduced the phosphorylation level of ERK and shortened the activation time, whereas the phosphorylation level of Akt remained unchanged. Collectively, our findings revealed the hetero-dimerization of CXCR4 and XCR1 and its effects on CXCR4-mediated cellular motility, receptor internalization, and ERK pathway phosphorylation. Therefore, XCR1-targeting drugs could be candidates for cross-desensitization of CXCR4 and might represent a possible option for inhibiting breast cancer metastasis.

Objective: To compare the effects of artesunate (Art) and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis. Methods: Eighty C57BL/6 female mice were randomly divided into healthy control group, infection group, Art treatment group and Fuzheng Huayu Decoction treatment group, with 20 mice in each group. Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae. After 6 weeks, praziquantel (300 mg/kg) was used for 2 days to kill the worms. The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day, while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day. After 6 weeks, fresh liver tissues of the four groups were collected. Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A (SDHA) and malate dehydrogenase (MDH2), citrate synthase (CS), ketoglutarate dehydrogenase (OGDH), and target of rapamycin 1 (mTORC1) pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues. The relative expression levels of adenylate activated protein kinase (AMPK) and mitochondrial autophagy pathway kinase (PINK1) were detected. Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate. Two-way ANOVA was used to compare the significance and difference between two sets of samples. Results: Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group. Western blot analysis showed that the infection group (0.82 ± 0.05) had significantly lower relative expression of SDHA protein than the healthy control group (1.00 ± 0.05) (t = 11.23, P = 0.0035), while the Art treatment group (0.73 ± 0.05) had significantly higher relative expression of SDHA protein than the infection group (t = 10.79, P = 0.0073). However, there was no significant change in Fuzheng Huayu Decoction treatment group (0.98±0.05) (t = 1.925, P = 0.1266). The relative expression of p-AMPK protein was significantly higher in the infection group (1.15 ±0.05) than in the healthy control group (0.98 ± 0.07, t = 12.18, P = 0.0029), and the expression of p-AMPK in the Art treatment group (0.50 ± 0.05) was significantly lower than the infection group (t = 11.78, P = 0.0032). The relative protein expression of AMPK was significantly lower in the infection group (0.80 ± 0.05) than in the healthy control group (1.00 ± 0.05, t = 10.53, P = 0.0046). The expression of AMPK was significantly lower in the Art treatment group (0.54 ± 0.05) than in the infection group (T = 13.98, P = 0.0036). The relative expression of p-mTORC1 protein (0.93 ± 0.08) was not significantly different in the infection group than in the healthy control group (t = 2.28, P = 0.065), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1 protein than the infection group (t = 10.58, P = 0.029). The expression of p-mTORC1/ m-TORC1 was not significantly different in the infection group (0.98 ± 0.03) than in the healthy control group (0.97 ± 0.03, t = 0.98, P = 0.085), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1/ m-TORC1 than the infection group (t = 14.58, P = 0. 009). The relative protein expression of PINK1 was significantly lower in the infection group (0.55 ± 0.05) than in the healthy control group (1.00 ± 0.03, t = 13.49, P = 0.0011), while the Art treatment group (1.21 ± 0.05, t = 9.98, P = 0.0046) and Fuzheng Huayu Decoction treatment group (1.31 ±0.35, t = 6.98, P = 0.027) had significantly higher relative protein expression of PINK1 than the infection group. Mitochondrial function tests showed that after adding substrate complex II, the oxygen consumption of the infection group was lower than the healthy control group, while the Art treatment group and the Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. The oxygen consumption was significantly lower after adding the substrate complex III in the infection group than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. Conclusion: Art can alleviate schistosomiasis liver fibrosis by inhibiting AMPK/mTORC1 signaling pathway activity and enhancing mitochondrial oxygen consumption, autophagy and SDHA expression.
Animals ; Artesunate ; Drugs, Chinese Herbal/therapeutic use* ; Female ; Liver Cirrhosis/drug therapy* ; Mice ; Mice, Inbred C57BL ; Mitochondria ; Schistosomiasis