OBJECTIVE To investigate whether quinolinic acid(QA)induces autophagy in PC12 cells and its relationship with glycogen synthase kinase-3β(GSK-3β)/β-catenin related signaling path?ways. METHODS PC12 cells were treated with QA 2.5,5.0 and 10.0 mmol·L-1 for 24 h. The cell viability was determined by MTT assay. Autophagy fluorescent spots labelled form of microtubule-associated protein 1 light chain 3(LC3)was examined by LC3 immunostaining. The expressions of GSK-3β,β-catenin,LC3 and Beclin 1 were determined by Western blotting. RESULTS QA inhibited PC12 cell survival in a concentration-dependent manner,and IC50 was 8.7 mmol · L- 1. Compared with normal control group,QA 2.5,5.0 and 10.0 mmol · L-1 increased autophagic intracellular LC3 fluorescence spots,elevated the expression ratio of LC3-Ⅱ/LC3-Ⅰ and expression of Beclin 1 in PC12 cells(P<0.05). In addition,QA enhanced GSK-3βexpression and decreasedβ-catenin expression(P<0.05,P<0.01). CONCLUSION QA induces autophagy in PC12 cells. This mechanism may be associated with the activation of GSK-3β/β-catenin related signaling pathways.

OBJECTIVEThis study investigated the thrombolitic effect of Tianma Xinnao capsule and its preliminary mechanism, as well as the effect on coagulation system.

METHODCharlton's and Tomihisa's methods were modified to investigate the thrombolytic effect of Tianma Xinnao capsule. The activities of type 1 plasminogen activator inhibitor (PAI-1) and tissue-type plasminogen activator (tPA) in rabbit plasma were assayed by use of ELISA. The effects of Tianma Xinnao capsule were also evaluated on euglobulin lysis time (ELT), prothrombin time (PT), kaolin partial thromboplastin time (KPTT), thrombin time (TT), fibrinogen (Fib) and hemorheology.

RESULTThe results showed that Tianma Xinnao capsule had a dose-dependent thrombolytic effect in rats. Tianma Xinnao capsule at 0.6 and 1.2 g x kg(-1) produced 40% and 50% of reperfusion rate, while obtained 50% and 40% of reocclusion rate; 0.3 g x kg(-1) of Tianma Xinnao capsule, however, had no effect on the reperfusion or reocclusion rate. Tianma Xinnao capsule significantly inhibited PAI-1 activity, while elevated tPA activity in rabbit plasma. Tianma Xinnao capsule markedly prolonged ELT, PT, and KPTT, and decreased Fib level. Tianma Xinnao capsule showed no significant influence on TT or high, medium, or low sheering viscosity.

CONCLUSIONIt is indicated that Tianma Xinnao capsule inhibited PAI-1 activity and increased tPA activity, and this property of Tianma Xinnao capsule is assigned to be responsible for the thrombolytic effect.

Animals ; Blood Coagulation ; drug effects ; Blood Viscosity ; drug effects ; Capsules ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rabbits ; Rats ; Thrombosis ; blood ; drug therapy ; metabolism ; Tissue Plasminogen Activator ; metabolism

Objective To explore the change of miRNA-16(miR-16) level in the patients with multiple my-eloma(MM ) and its relationship with the Mayo risk stratification and prognosis judgment .Methods Each 10 mL of bone marrow samples was collected from 31 cases of MM in the hematology department of the First Af-filiated Hospital of Guangxi Medical University from August 2016 to January 2017 ,and each 5 mL of bone marrow sample was collected from 5 healthy bone marrow donors as the healthy control .The real time fluores-cence quantitative PCR(qRT-PCR) was used to analyze relative the change of miR-16 relative expression level and fluorescence in situ hybridization(iFISH) was used to detect the abnormal gene .Then the relationship be-tween miR-16 relative expression level with Mayo based risk stratification ,and the difference of miR-16 rela-tive expression level between before and after self treatment were statistically analyzed .Results The miR-16 relative expression level in MM patients was increased and positively correlated with the Mayo risk stratifica-tion .Meanwhile in the MM patients with disease progression ,the miR-16 expression was increased . Conclusion miR-16may serve as the indicator for judging the recurrence or progression of MM ,and guides the clinical personalized therapy of MM according to its expression level change .

Objective: To evaluate the clinical observation of rectal prolapse by partial perineal rectal sigmoidectomy. Methods: Sixty patients with rectal prolapse who met the diagnostic criteria of rectal prolapse in Yiwu Central Hospital from April 2013 to June 2015 were selected as the research subjects.They were divided into two groups according to random number table, with 30 cases in each group.The observation group used Altemeier, and the control group underwent triple surgery (rectal mucosal ligation, periorbital injection, and anal ring retraction). The total effective rate after surgery was compared and analyzed. Results: The operation of the two groups was successfully completed.The total effective rate of the observation group was 96.67%, which was significantly higher than 80.00% of the control group, the difference was statistically significant(χ²=5.856, P=0.048). The Wexner constipation score of the observation group was (1.4±1.6)points, which was significantly lower than that of the control group[(3.2±2.0)points, t=3.849, P<0.05]. The Wexner anal incontinence score of the observation group after 24 months was (2.9±1.8)points, which was significantly lower than that of the control group [(4.4±2.7)points, t=2.532, P<0.05]. The proportions of anastomotic bleeding, pelvic swelling and perianal eczema in the observation group were 0.00%, 0.00% and 3.33%, respectively, which were significantly lower than those in the control group(20.00%, 23.33% and 30.00%, χ²=4.630, 5.822, 7.680, all P<0.05). Conclusion Altemeier can significantly improve the quality of life of patients, with a high cure rate and good clinical results.

Objective: To explore the difference of liver inflammation and fibrosis in patients with chronic hepatitis B virus (HBV) infection and chronic hepatitis C virus (HCV) infection, and to investigate the relationship between hepatic pathology and alanine aminotransferase (ALT). Methods: 57 patients with chronic HCV infection and 346 patients with chronic HBV infection who were hospitalized at Shengjing Hospital of China Medical University from January 2012 to September 2016 were enrolled. In chronic HBV infection, including 88 cases whose ALT were more than two times of upper limited of normal (ALT≥2×ULN) and 258 cases whose ALT were less than two times of upper limited of normal (ALT < 2×ULN).All the patients were underwent liver biopsy. Chronic HBV infection (ALT≥2×ULN and ALT < 2×ULN) and chronic HCV infection were compared respectively. Statistical analyses were performed using a Univariate χ²-test and Mann–Whitney U test for comparison. Correlations between variables were analyzed using Spearman's rank correlation. Results: In chronic HBV infection group, 169 cases (48.8%) had inflammation grade≥2 (G≥2), 98 cases (28.3%) had fibrosis stage≥2 (S≥2), 81 cases (23.4%) with G≥2 and S≥2.In the ALT < 2×ULN group, there were 109 cases (42.2%) with G≥2, 62 cases (24%) with S≥2, 49 cases (19%) with G≥2 and S≥2. In the ALT≥2×ULN group, 60 cases (68.2%) with G≥2, 35 cases (39.8%) with S≥2, 31 cases (35.2%) with G≥2 and S≥2. The grade of inflammation and fibrosis have significantly different between ALT≥2×ULN group and ALT < 2×ULN group (χ² = 17.66, χ² = 8.06, P < 0.01). In chronic HCV infection group, 47 cases (82.5%) with G≥2, 20 cases (35.1%) with S≥2, 20 cases (35.1%) with G≥2 and S≥2. ALT had no correlation with inflammation and fibrosis (P > 0.05). The grade of inflammation was significantly different between chronic HCV infection and chronic HBV infection whose ALT < 2×ULN (χ² = 30.19, P < 0.01) but the fibrosis have no difference (χ² = 2.96, P > 0.05). Compared with chronic HBV infection whose ALT≥2×ULN, both inflammation and fibrosis had no significantly different (χ² = 3.65, χ² = 0.32, P > 0.05 respectively). Conclusion In chronic HBV infection whose ALT < 2×ULN, about 30%-40% liver tissue with significant necroinflammation and /or fibrosis. About 80% chronic HCV infection with significant necroinflammation, and the grade of inflammation has no correlation with ALT. The grade of inflammation has significantly different between chronic HCV infection group and chronic HBV infection group whose ALT < 2×ULN.

Precision medicine is an emerging medical strategy that takes into account individual molecular variations of disease to guide accurate prevention and treatment. Tumor molecular markers closely related to aggressive behavior and treatment response will be identified by integrating clinical information and multiple?omics, and will be verified in well?designed clinical trials. Breast cancer is a highly heterogeneous disease. Its treatment based on molecular subtyping has achieved initial success. However, drug resistance and tumor heterogeneity are still major challenges. This review will focus on the recent progress and future prospects of clinical research on breast cancer in the era of precision medicine.

Precision medicine is an emerging medical strategy that takes into account individual molecular variations of disease to guide accurate prevention and treatment. Tumor molecular markers closely related to aggressive behavior and treatment response will be identified by integrating clinical information and multiple?omics, and will be verified in well?designed clinical trials. Breast cancer is a highly heterogeneous disease. Its treatment based on molecular subtyping has achieved initial success. However, drug resistance and tumor heterogeneity are still major challenges. This review will focus on the recent progress and future prospects of clinical research on breast cancer in the era of precision medicine.

OBJECTIVE：To evaluate the dissolution behavior consistency between the generic drugs and original drugs of Oxcarbazepine scored tablets ，and to compare the appearance ，the friability of the split portions ，loss of mass of the split portions as well as crystal form and morphology of raw material from different enterprises. METHODS ：HPLC method was adopted. The paddle method （rotation speed of 60 r/min，the temperature of 37.0℃）was adopted to determine accumulative dissolution rate of generic and original drugs in 4 mediums [ 0.6％ SDS hydrochloric acid solution （pH＝1.2），0.6％ SDS acetate buffer solution （pH＝4.5），0.6％ SDS phosphate buffer solution （pH＝6.8）and 0.6％ SDS water solution]. The similarity factor method was used to evaluate the similarity of dissolution curves as well as intra-batch uniformity of the split portions and whole tablets. The friability tester and electronic balance were used to determine the friability and the loss of mass of the split portions. X-ray diffractometer and scanning electron microscope were used to observe the crystal form and crystal morpho logy of the raw materials of different enterprises. RESULTS ：The linear range of oxcarbazepine was LOD was 0.04 μg/mL；RSDs of precision ，stability，reprodu- cibility and durability tests were lower than 2.0％；the reco- veries were 99.80％-101.63％（RSD＝0.37％-0.91％，n＝3）. The average cumulati ve dissolution rate of generic drug A ， generic drug B and original drug in 4 different dissolution media at 90 min were 92％，87％，90％ [0.6％ SDS hydrochloric acid solution（pH＝1.2）]；94％，94％，90％ [0.6％ SDS acetate buffer solution （pH＝4.5）]；95％，95％，91％ [0.6％ SDS phosphate buffer solution （pH 6.8）]；97％，98％，95％（0.6％ SDS water solution ）. The similarity factors of generic drug A ，generic drug B and original drug in 4 kinds of different dissolution media were 66 and 81，71 and 69，71 and 61，59 and 39. In the first 15 min，the difference of dissolution rate of split portions and whole tablets were －3％-13％，－2％-24％ and －3％-7％ for generic drug A ， generic drug B and original drug ，respectively. RSDs of accumulative dissolution rate of split portions and whole tablets were 6％-14％ and 2％-9％ for generic drug A （n＝12），4％-10％ and 1％-8％ for generic drug B （n＝12）and 2％-7％ and 2％-8％ for original drug. The appearance of the original drug was fusiform ，and the notch was deep ；the shape of the generic drug was different from each other ，and the notch of the generic drug was significantly shallower than that of original drug. The friability ， the loss of mass of the split portions for generic drug A and generic drug B ，original drug were 0.62％and 0.67％，0.12％ and 0.11％，0.08％ and 0.05％. The domestic raw materials possessed irregular lumps and debris ，while the raw materials produced by original drug enterprises possessed regular flat cuboids and regular strips with little debris ；but X-ray diffraction peaks of them were basically the same. CONCLUSIONS ：The dissolution behavior of generic drug A in 4 medium is consistent with that of the original drug；dissolution behavior of generic drug B in water containing 0.6％SDS is different from that of the original drug ；there is no significant change in the homogeneity of the original drug before and after splitting ，but the homogeneity of the generic drug A and B after splitting is lower than that of the whole tablet ；the fragility of generic drugs and loss of mass of split portions are higher than those of the original drugs ；two kinds of raw material have the same crystal form but different crystal morphology.

Objective: To buildSERPING1-knockout porcine fetal fibroblasts(PFFs) based on CRISPR/Cas9 technology and provide cell experimental materials for the construction of hereditary angioedema models. Methods: Firstly, protein structure prediction software was used to analyze the amino acid homology between human and pigSERPING1. Secondly, the eighth exon was selected as the knockout target by screening the effective coding region of the pigSERPING1gene. A single guide RNA targeting pigSERPING1was designed, synthesized and linked to pX330 vector containing Cas9 endonuclease. G418 was used to obtain the positive monoclonal cells after transfection into PFFs. Finally, the circumstance of CRISPR/Cas9 mediated knockout was assessed by the T7 EN1 enzyme digestion assay and the genotypes of monoclonal cells were identified by sequencing analysis Results: Bioinformatic analysis revealed that theSERPING1protein of human and pig had high homology, amino acid sequence identity reached 65.87%. A vector targetingSERPING1was constructed successfully and transfected into cells. Monoclonal cells with knockoutSERPING1gene were obtained through drug screening. Sequencing confirmed the mutant genotype. Conclusion The human and pigSERPING1sequences and their protein structures are highly homologous, and it is suitable for the construction of disease model. CRISPR/Cas9 expression vectors were constructed to achieve theSERPING1gene targeting in PFFs.SERPING1-knockout monoclonal cells were obtained, which could contribute to the construction of theSERPING1knockout miniature pig model.

OBJECTIVE：To es tablish a method for the content determination of heavy metals [lead （Pb），cadmium（Cd）， copper （Cu）， mercury （Hg）] and harmful elements [arsenic （As）] in Pediatric paracetamol artificial cow-bezoar and chlorphenamine maleate granules. METHODS ：The samples were conducted pretreatment by microwave digestion instrument and determined by inductively coupled plasma mass spectrometry （ICP-MS）using elements germanium ，indium，bismuth as internal standard. RESULTS ：The linear ranges of Pb ，As，Cu，Cd and Hg were 1-20，0.5-10，5-100，0.5-10 and 0.2-4 ng/mL， respectively （all r＞0.997）. The limits of detection （LODs） were 0.041 1，0.013 2，0.057 3，0.009 0，0.005 4 ng/mL， respectively. The limits of quantification （LOQs）were 0.137 0，0.044 0，0.191 0，0.030 0，0.018 0 ng/mL，respectively. RSDs of precision and repeatability tests were all less than 6％. RSDs of stability tests （28 h）of Pb ，As，Cu and Cd were all less than 5％， and that of stability test （28 h）of Hg was less than 7％. The average recoveries were 89.44％（RSD＝5.87％，n＝9），99.56％ （RSD＝5.46％ ，n＝9），96.12％（RSD＝4.62％ ，n＝9），105.82％（RSD＝2.80％ ，n＝9）and 90.23％（RSD＝3.59％ ，n＝9）， respectively. Five elements were all detected in 63 batches of samples ，and the contents of them were 0.191 0-1.527 6，0.002 5- 0.047 4，0.034 1-1.549 0，0.001 5-0.078 8 and 0.001 9-0.005 4 mg/kg，respectively. CONCLUSIONS ：The method is simple ， sensitive and accurate. It is suitable for simultaneous determination of 5 elements in Pediatric paracetamol artificial cow-bezoar and chlorphenamine maleate granules.