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 observe the neural protection of 3-n-butylphthalide (NBP) injection in focal cerebral ischemia-reperfusion rats. Methods 160 male Sprague-Dawley rats were randomly divided into sham group (n=10), ischemia-reperfusion group (IR group, n=50), high-dose NBP treatment group (high-dose group, n=50), middle-dose NBP treatment group (middle-dose group, n=25) and low-dose NBP treatment group (low-dose group, n=25). The later 4 groups were occluded the middle cerebral artery for 2 hours and reperfused. The sham group was sacrificed 24 hours after operation, and the other groups at 6, 12, 24, 48 and 72 hours after reperfusion, in which 5 of them were stained with TdT mediated dUTP Nick End Labeling (TUNEL) to observe the neuronal apoptosis, and immunohistochemistry to observe the expression of silent information regulation 2 homolog 1 (SIRT1) and peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α);the other 5 of sham group, IR group and high-dose group were observed with quantitative real-time PCR of SIRT1 and PGC-1α. Results Compared with the IR group, the number of apoptotic cells decreased and the SIRT1 and PGC-1αpositive cells increased in all NBP groups at each time (F>160.60, P<0.001), and it was the least of apoptotic cells and most of SIRT1 and PGC-1α positive cells in the high-dose group (P<0.05), while there was significant difference between the low-dose group and the middle-dose group, excluding 6 hours after reper-fusion (P<0.05). Compared with IR group, the expression of SIRT1 and PGC-1αmRNA increased in the high-dose group at each time (t>4.13, P<0.01). Conclusion NBP can protect brain from apoptosis in focal cerebral ischemia-reperfusion rats, which may relate to more ex-pression of SIRT1 and PGC-1α.

Objective To evaluate clinical application value of polymerase chain reaction (PCR) detection for bacteria in peritoneal dialysis associated peritonitis (PDAP).Methods Peritoneal dialysis fluid specimens were collected from January 2014 to December 2014 in The First Affiliated Hospital of Anhui Medical University.Conventional bacterial culture and PCR detection were used respectively.According to the bacterial 16S rRNA gene, universal primers were devised and designed, based on reference, the specific primers of 17 kinds of experimental bacteria.Real-time fluorescent PCR (Real-time PCR, qPCR) amplification was implemented.The establishment of standard strain DNA extract was used as positive control;sterile double distilled water was used as negative control.Results (1) The traditional bacterial culture results showed that positive proportion was 26/40 in specimen of 40 cases, gram-positive strains accounting for 18/26.Main species were epidermis staphylococcus (5/26), hemolysis staphylococcus (4/26), escherichia coli (4/26), and streptococcus viridans (3/26).(2) The PCR detection results showed that total positive rate was 33/40 in 40 patients specimens, among which 2 cases of positive samples ended up with no specific strains being detected;the main bacteria strains in PCR were not different from ordinary culture results.(3) With bacterial culture as the gold standard, the detection sensitivity of PCR technology for PDAP pathogenic bacteria was 96.15％ and specificity was 42.86％;the detection positive rate was significantly higher than ordinary culture method.(4) PCR technology for detecting pathogenic bacteria could produce results within 4-6 hours, while reported positive results in the traditional bacterial culture would take (77.88±15.53) hours, which was significantly longer than PCR.Conclusion Compared with traditional bacteria culture method, PCR method is more sensitive, simple, and quick.Bacteria detection using PCR technique is of clinic applied value in PDRP.

[Abstract ] Objective The purpose of this study was to observe the effects of dl-3n-butylphthalide (NBP) sodium chloride injection post-processing on the expressions of X-inhibitor of apoptosis (XIAP) and Bcl-2/adenovirus E1B19kDa interacting protein 3 (BNIP3) in the hippocampus CA1 neurons of focal cerebral ischemia reperfusion (IR) rats, and to investigate the brain-protection mechanisms of NBP. Methods A total of65 adult male Sprague-Dawley rats were divided into five groups of equal number, sham op-eration, IR, and low-,medium -and high-dose NBP, according to the random number table. The IR models were established by modified ligation of the middle cerebral artery.The animals in the NBP groups received intra-abdominal injection of NBP at 2, 4, and 6 mg/kg, re-spectively.All the rats were sacrificed at 24 hours after modeling,neurological scores obtained by Zea Longa, the volume of infarction measured by TTC staining, the number of apoptotic cells counted by TUNEL, and the expressions of XIAP and BNIP3 detected by immunohistochemistry and real-time PCR. Results The neural function defect scores were markedly lower in low-, medium-and high-dose NBP groups than in IR model rats (P<0.05), with statis-tically significant differences among the three dose groups (P<0.05).The volume of infarction was remarkably higher in the low-dose than in the medium-and high-dose NBP groups (P<0.05).The number of apoptotic cells in the hippocampus CA1 neurons was de-creased in the NBP groups as compared with the IR models (P<0.05).The XIAP-and BNIP3-positive cells were significantly in-creased in the IR model rats as compared with the sham operation group ([22.31 ±0.94] and [60.13 ±2.59]/HP vs [3.07 ±1.43] and [5.78 ±0.44]/HP, P<0.05).In comparison with the IR models, the NBP-treated rats showed a progressively increased number of XIAP-positive cells in low-, medium-, and high-dose groups ([28.70 ±1.18], [32.79 ±0.88], and [37.01 ±1.24]/HP) (P<0.05) but a decreased number of BNIP3-positive cells in the three dose groups ([52.07 ±1.02], [40.30 ±2.00], and [31.04 ± 0.43]/HP) (P<0.05).Similarly, the expression of XIAP mRNA was up-regulated while that of BNIP3 mRNA down-regulated in the NBP treatment groups as compared with the IR model rats, both in a dose-dependent manner (P<0.05). Conclusion NBP post-processing has a neuroprotective effect on IR rats, which is associated with its impact on the expressions of XIAP and BNIP3.

The serum level of CXCL10 was determined in both patients with Graves' disease (GD) and normal control subjects.Serum CXCL10 levels in untreated and relapsed GD groups were higher compared with the remission group and control group( P＜0.01 ).A significant difference was observed between the former two GD groups (P＜0.01 ),but no difference was found between latter two groups( P＞0.05 ).Serum CXCL10 levels were positively correlated with FT3 and FT4,and negatively correlated with TSH by multiple linear regression analysis( P＜0.01 ).

Objective To investigate the pathogens and their antibiotic resistance in patients with peritoneal dialysis‐related peritonitis .Methods The clinical data including pathogens ,antibiotic resistance profile of 213 patients with peritoneal dialysis‐related peritonitis who were treated in our peritoneal dialysis center from January 2011 to December 2013 were analyzed retrospectively .Results Dialysate culture was positive for 132 (62 .0% ) of the 213 cases ,resulting in a total of 140 strains of microorganisms ,including 84 strains of gram positive cocci ,37 strains of gram‐negative bacilli ,10 strains of fungus and 9 strains of gram positive bacilli . Coagulase‐negative Staphylococcus was the most common gram positive bacteria while Escherichia coli was the most common gram negative bacteria isolated from the effluent .The prevalence of methicillin‐resistant S .aureus and methicillin‐resistant coagulase negative Staphylococcus was 14 .3% (1/7) and 43 .2% (19/44) ,respectively . About 44 .4% (8/18) of the E .coli and K . pneumoniae isolates produced extended spectrum beta‐lactamases .All the gram‐positive cocci were sensitive to vancomycin and linezolid and slightly resistant to chloramphenicol (6 .3% ) , moxifloxacin (8 .5% ) , and rifampicin (9 .5% ) , but highly resistant to cefazolin (90 .0% ) ,followed by ampicillin (76 .7% ) ,oxacillin (71 .2% ) and penicillin (69 .7% ) . Coagulase negative Staphylococcus isolates were sensitive to vancomycin , linezolid , tigecycline , quinupristin‐dalfopristin and daptomycin ,but all resistant to cefazolin and ampicillin ,and highly resistant to penicillin (91 .9% ) and oxacillin (82 .5% ) .All the gram‐negative bacilli were sensitive to meropenem ,ertapenem ,cefoperazone‐sulbactam and tigecycline .About 80 .6% and 65 .5% of the gram‐negative bacilli were resistant to ampicillin and peperacillin ,respectively .E .coli isolates were sensitive to meropenem ,ertapenem and piperacillin‐tazobactam but highly resistant to ampicillin (81 .3% ) and piperacillin (71 .4% ) . Conclusions Gram‐positive cocci especially Staphylococcus and gram negative bacteria E .coli are major pathogens in peritoneal dialysis‐related peritonitis .Adequate microbiological culture and suitable antimicrobial therapy are key to successful treatment of the peritonitis associated with peritoneal dialysis .

Seventy-three patients with type 2 diabetes mellitus were divided into atheroselerosis(AS) group and non-AS group according to the intima-media thickness(IMT)of the carotid artery.The plasma visfatin level in AS group was higher than that in non-As group[(44.95±10.14 vs 34.52±9.08)μg/L,P<0.05],and both of them were higher than that of the control [(24.46±7.18)μg/L,both P<0.05 ].The visfatin level Was positively correlated with IMT,waist-to-hip ratio,visceral fat thickness,fasting insulin,and HOMA insulin resistance index.Age,duration of diabetes,HbA_(1C),and visfatin level were the major risk factors for IMT of the carotid artery.

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.

BACKGROUNDHyperglycemia may accelerate liver fibrosis. Currently, there is no effective treatment for liver fibrosis induced by type 2 diabetes. The study aim was to investigate whether RhoA/Rho kinase (ROCK) pathway is involved in liver fibrosis in the rats with type 2 diabetes and define the protective effects of fasudil on livers.

METHODSA rat model of type 2 diabetes was established by high fat diet combined with streptozotocin (30 mg/kg, intraperitoneal injection). Animals were randomly assigned to 3 groups: control rats, untreated diabetic rats that received vehicle and fasudil-treated diabetic rats that received ROCK inhibitor fasudil hydrochloride hydrate (10 mg/kg per day, intraperitoneal injection, for 14 weeks). The morphological features of liver were observed by HE staining. Accumulation of collagen in livers was determined by Masson staining and the measurement of hydroxyproline. The mRNA expression of transforming growth factor-β1 (TGFβ1), connective tissue growth factor (CTGF), type-I, and type-III procollagen was assessed with real-time polymerase chain reaction. The phosphorylation of myosin phosphatase target subunit-1 (MYPT1) and the protein levels of TGFβ1 and α-smooth muscle actin (a-SMA) were evaluated by Western blotting.

RESULTSCompared with control rats, untreated diabetic rats showed higher values of collagen and hydroxyproline in livers (P < 0.01), the phosphorylation of MYPT1 and the protein levels of TGFβ1 and α-SMA were increased (P < 0.01), and the mRNA expression of TGFβ1, CTGF, type-I, and type-III procollagen was upregulated (P < 0.01); compared with untreated diabetic rats, treatment with fasudil signifcantly reduced values of collagen and hydroxyproline (P < 0.01), and decreased the phosphorylation of MYPT1 and the levels of TGFβ1 and α-SMA (P < 0.01), concomitant with the downregulation of TGFβ1/CTGF, type-I, and type-III procollagen mRNA expression (P < 0.01).

CONCLUSIONSFasudil ameliorates liver fibrosis in rats with type 2 diabetes at least partly by inhibiting TGFβ1/CTGF pathway and α-SMA expression. Inhibition of RhoA/ROCK may be a novel therapeutic target for liver fibrosis in diabetic non-alcoholic steatohepatitis.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine ; analogs & derivatives ; therapeutic use ; Animals ; Connective Tissue Growth Factor ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; Female ; Liver Cirrhosis ; drug therapy ; enzymology ; metabolism ; Protein Kinase Inhibitors ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; rho-Associated Kinases ; antagonists & inhibitors