OBJECTIVE: To assess whether adjuvant Chinese patent medicines (CPMs) to standard treatment could reduce recurrent bleeding after variceal bleeding in cirrhotic patients. METHODS: This study retrospectively collected 555 consecutive patients who recovered from variceal bleeding. A population-based cohort study was established depending on if adjuvant CPMs were administered to prevent rebleeding. A total of 139 patients who had taken ⩾28 cumulative defined daily doses (cDDDs) of CPMs were included in the CPMs cohort, and 416 patients who used <28 cDDDs of CPMs were enrolled in the non-CPMs cohort. On evaluation of rebleeding incidence, 1:2 propensity score matched was used to estimate for reducing bias. Patients were followed for at least 12 months. The end-point of this study was clinically significant esophagogastric variceal rebleeding. RESULTS: Following multivariate analysis, CPMs therapy was an independent factor for variceal rebleeding [adjusted hazard ratio (AHR)=0.657; 95% confidence interval=0.497-0.868; P=0.003]. After the 1:2 propensity score matching, a significant reduction (23.5%) in the incidence of variceal rebleeding in patients was observed, from 58.3% in the non-CPMs cohort to 44.6% in the CPMs cohort (modified log-rank test, P=0.002) within a year. The AHRs for rebleeding were 0.928, 0.553, and 0.105, for 28-90 cDDDs, 91-180 cDDDs, and >180 cDDDs of CPMs, respectively. The median rebleeding interval in the CPMs cohort was significantly larger compared with the non-CPMs cohort (113.5 vs. 93.0 days; P=0.008). CONCLUSION Adjuvant CPMs to standard therapy can significantly reduce the incidence of variceal rebleeding and delay the time to rebleeding.

AIM:To investigate the expression and localization of autophagy related protein microtublule associated protein 1 light chain 3 (LC3) at various stages of follicular development and atresia in the mice.METHODS:On 0,1,2,3,4 and 5 day after intraperitoneal injection of pregnant mare serum gonadotropin (PMSG),expression and positioning situation of autophagy related protein LC3 and apoptosis related protein cleaved caspase-3 were examined by the method of immunohistochemical staining.The protein levels of cleaved caspase-3 and LC3 were determined by Western blot in cultured mouse granulosa cells after incubation under serum-free conditions in the absence or presence of FSH.LC3 subcellular localization in granulosa cells were studied by the method of immunofluorescence.RESULTS:The LC3 protein expressed in granulosa cells during all developmental stages mainly.Granulosa cells of atretic follicles that showed intense staining of cleaved caspase-3 and LC3.The protein levels of cleaved caspase-3 and LC3-Ⅱ in the granulosa cells significantly decreased at 1 d and 2 d after intraperitoneal injection of PMSG (P ＜ 0.05).The protein levels of cleaved caspase3 and LC3-Ⅱ in the granulosa cells increased in turn on 3,4 and 5 day after intraperitoneal injection of PMSG.The positive correlation between LC3-Ⅱ and cleaved caspase-3 protein levels was observed (r2 =0.8299,P ＜ 0.05).The LC3-Ⅱ protein expressed with punctuate structures in granulosa cell cytoplasm cultured under serum-free conditions in the presence of FSH.CONCLUSION:LC3 is expressed in the follicular granulosa cells with cell specificity and regional specificity.Autophagy is induced mainly in granulosa cells during folliculogenesis and shows positive correlation with apoptosis.Ovarian granulosa cell autophagy and apoptosis are gonadotropic hormone dependent.

Objective To investigate the difference of late-phase of limb ischemia preconditioning (L-LIP) verse early-phase (E-LIP) on patients with percutaneous coronary intervention (PCI).Methods A total of 160 patients with unstable angina pectoris who were planned to undergo PCI were divided equally into two groups at random.The late-phase of limb ischemia preconditioning group (80 patients) were provided with L-LIP (three 5-minute inflations up to 200mmHg by applying the sphygmomanometer cuff around the right upper arm,followed by 5-min intervals of reperfusion,twice a day) 3 days before PCI.The Earlyphase of limb ischemia preconditioning group (80 patients) were provided with E-LIP (method as above)2 hours before PCI.Comparison of procedural parameters during PCI and the levels of cTnT,CK-MB,hs-CRP were made 24 hours after PCI.Estimation of the rate of adverse events at 1 year between the two groups was evaluated by Kaplan-Meier analysis.Results Compared to the E-LIP group,the rates of angina,arrhythmia and TIMI flow ≤ 2 during PCI were significantly lower in the L-LIP group (all P ＜ 0.05).At 24 hours after PCI,the levels of cTnT and CK-MB were declined more significantly in the L-LIP group[(11.52±2.41) pg/ml vs.(27.53±4.78)pg/ml,P =0.021;(14.11±2.87)Iu/L vs.(30.23±5.17)Iu/L,P =0.032].There was no difference in the level of hs-CRP between the 2 groups [(128±0.71)mg/dl vs.(1.33±0.69)mg/dl,P =0.742].The Kaplan-Meier survival curve showed that the incidence rate of adverse events in the L-LIP group at l year was lower than the E-LIP group (3.75％ vs.13.75％,P =0.024).Conclusions L-LIP is more effective to in protecting myocardial cell in patients with unstable angina pectoris undergoing elective PCI and may reduce the rate of future adverse event.

OBJECTIVETo establish a rapid and convenient method of DNA modification by bisulfite sodium for the detection of DNA methylation.

METHODSThrough increasing the bisulfite sodium concentration and the temperature of treatment, cutting down the modification time, besides using glassmilk to adsorb the DNA in the purification and recovery, to improve the methods of DNA modification. Efficiency of cytosine converted to thymine in MAGE-A3 gene and DAP-K gene fragments were analyzed by bisulfite sequencing PCR in order to evaluate the DNA modification effect among the improved method, traditional method and kit method.

RESULTSThe operating time of test was shortened to about 3 hours by the improved method; conversion rate of unmethylated cytosine to thymine was over 99%; compared with the traditional method and kit method, there was no significant difference (χ(2) = 0.0564, P > 0.05); the improved method was only for the unmethylated cytosine conversion modification, and there was no significant difference in process of methylated cytosine converted to thymine comparing with the traditional method (χ(2) = 0.0149, P > 0.05).

CONCLUSIONThe improved method has high efficiency of DNA modification and has no significant effect on excessive modification;meanwhile, it has many advantages such as time-saving and easy to operate etc.

Cytosine ; chemistry ; DNA ; chemistry ; DNA Methylation ; Polymerase Chain Reaction ; Sulfites ; chemistry ; Thymine ; chemistry

OBJECTIVETo evaluate the clinical efficacy and safety of integrative medical program based on blood cooling and detoxification recipe (BCDR) in treating patients with hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) of heat-toxicity accumulation syndrome (HTAS).

METHODSAdopting randomized controlled clinical design, a total of 105 HBV-ACLF patients of HTAS were randomly assigned to the trial group (64 cases) and the control group (41 cases). Patients in the control group were treated with comprehensive Western therapy, while those in the trial group were treated with comprehensive Western therapy plus BCDR. All were treated for 8 weeks and followed up for 40 weeks. Effect and safety of the treatment were assessed, including fatality, liver functions [total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), and aspartate transaminase (AST)], and prothrombin activity (PTA) after treatment and at week 48 of follow-ups.

RESULTSAfter 8-week treatment, there was statistical difference in the overall fatality rate (15.63% vs 34.15%), the fatality rate in the mid-term (25.0% vs 64.7%), TBIL at week 8 (64.54 +/- 79.75), AST [at week 2: (178.97 +/- 44.24) U/L vs (288.48 +/- 58.49) U/L; at week 4: (61.65 +/- 27.36) U/L vs (171.12 +/- 89.11) U/L] and PTA [at week 4: (58.30 +/- 15.29) vs (42.56 +/- 15.27); at week 6: (60.77 +/- 20.40) vs (43.08 +/- 12.79)] (all P < 0.05). At week 48 of the followup, the fatality rate of the trial group (21.88%) decreased by 17. 14% when compared with that of the control group (39.02%; P < 0.05). No obvious adverse event occurred in the two groups during the 8-week treatment period.

CONCLUSIONBCDR could significantly reduce the mortality of HBV-ACLF patients.

Acute-On-Chronic Liver Failure ; drug therapy ; virology ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; End Stage Liver Disease ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Young Adult

Acupuncture Points ; Acupuncture, Ear ; instrumentation ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Spinal Nerve Roots ; physiopathology ; Spondylosis ; physiopathology ; therapy ; Young Adult

OBJECTIVETo analyze CYP17A1 gene mutations in a child patient with 17 alpha-hydroxylase/17, 20-lyase deficiency (17OHD), and to review characteristics of CYP17A1 gene mutations in Chinese patients with 17OHD.

METHODSClinical data were collected. PCR and DNA sequencing were performed to detect mutations in the patient.

RESULTSThe patient has presented classical features of 17OHD including hypertension, hypokalemia, decreased sex hormones and plasma cortisol, and elevated blood adrenocorticotrophic hormone. A compound heterozygous mutation c.987C>A and c.985del was detected in the CYP17A1 gene, which resulted in two premature stop codons at positions 328 and 417.

CONCLUSIONA compound mutation, c.987C>A and c.985del, has been identified in a patient with 17OHD. Among CYP17A1 gene mutations identified in Chinese patients, missence mutations have been most common, and exons 5 and 8 have been the mutation hotspots.

Adolescent ; Adrenal Hyperplasia, Congenital ; enzymology ; genetics ; Base Sequence ; Female ; Humans ; Lyases ; deficiency ; genetics ; Molecular Sequence Data ; Mutation ; Steroid 17-alpha-Hydroxylase ; genetics

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Anxiety Disorders ; therapy ; Female ; Humans ; Male ; Middle Aged ; Qi ; Young Adult

OBJECTIVETo screen the differentially expressing genes between silicotic lung tissue and normal lung tissue, to identify the differentially expressing genes of matrix metalloproteinase-12 (MMP-12) and Cathepsin E and to explore the roles of those genes in silicosis development.

METHODSThirty male SD rats were divided randomly into two groups: control group (6 rats) and exposure group (24 rats) which was exposed to SiO2 by intra-tracheal perfusion. On the 30 th, 60 th and 90 th days after exposure, 8 rats in model group and 2 rats in control group were executed and the lung tissues were obtained. The morphologic changes of lung tissues were observed with HE staining and VG staining under a light microscope. The gene microarrays were used to identify differentially expressing genes of lung tissues in rats exposed to SiO2 for 60 days. Two significantly up-regulated genes, MMP-12 and Cathepsin E, were validated using RT-PCR, immunohistochemistry and Western Blot assay.

RESULTSA total of 338 differentially expressing genes were identified from the 26 962 genes between silicotic rats and normal rats, including 267 up-regulated genes and 71 down-regulated genes. The results of RT-PCR showed that in the lung tissues of exposure group on the 30 th, 60 th and 90 th days, the mRNA expression levels of MMP-12 were 4.306, 5.338, 6.713 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.434, 2.974, 3.889 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the mRNA expression levels of MMP-12 were 1.435, 1.746, 2.069 times higher than those in the control group, the mRNA expression levels of Cathepsin E were 1.372, 1.663, 2.103 times higher than those in the control group, respectively. The results of immunohistochemical showed that in the lung tissues of exposure group on the 30th, 60th and 90th days, the expression levels of MMP-12 protein were 1.214, 1.531, 1.959 times higher than those in the control group, the expression levels of Cathepsin E protein were 1.262, 1.828, 1.907 times higher than those in the control group, respectively. Compared with the control group, the mRNA and protein expression levels of MMP-12 and Cathepsin E in lung tissues of exposure group were significantly up-regulated (P < 0.05).

CONCLUSIONThe differentially expressing genes in rat lung tissues screened by gene chip were validated, which suggested that a complex gene regulatory network may be contributed to occurrence of silicosis. MMP-12 and Cathepsin E genes may be involved in the development of silicotic pulmonary fibrosis by degrading the basement membrane of alveolar wall and participating in the immune response.

Animals ; Cathepsin E ; genetics ; metabolism ; Gene Expression ; Lung ; metabolism ; Male ; Matrix Metalloproteinase 12 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Silicosis ; genetics ; metabolism

Adenine ; analogs & derivatives ; therapeutic use ; Adult ; Bone Density ; drug effects ; Creatinine ; blood ; Hepatitis B, Chronic ; complications ; drug therapy ; metabolism ; Humans ; Male ; Organophosphonates ; therapeutic use ; Pain ; etiology ; Phosphorus ; blood