Suppression subtractive hybridization(SSH) is a good and highly effective method for gene cloning,which posses some character,such as convenient operation,high specificity,low background,good replication etc,and it can separate lower specific fragment.This article principally introduced its main technical points,strength and shortcomings and improvement of SSH,and also reviewed its applications in detecting the effects of radiation,heat,hypoxia,coldness and other environmental factors on gene expression.

Objective To explore the effect of laparoscopic bile duct exploration,balloon dilation,and catheter drainage in the treatment of postoperative recurrent choledocholithiasis.Method The data of 61 patients with postoperative recurrent bile duct stones from August 1999 to August 2009 were retrospectively analyzed.The patients received laparoscopic bile duct exploration (LCDE),laparoscopic papillary balloon dilation (LPBD),and laparoscopic papillary balloon dilation endoprosthesis (LPBDE).ResultSatisfactory outcome was achieved in all the 61 patients.Among the 61 patients,20 patients received LPBD and primary suturing,36 patients received LPBDE and primary suturing,and 5 patients received LPBDE and T-tube drainage.There was no residual stone.There was no perioperative mortality or serious complications including biliary leak and hemobilia.On follow up for 1 to 9.5 years in 56 patients,there was no recurrent choledocholithiasis.ConclusionThe procedures were feasible and safe,and they prevent recurrence of choledocholithiasis.

This paper reviewed the researches of TCM syndromes of rheumatoid arthritis in the past four years.The author believed that more and deeper epidemiological surveys are needed for studying rheumatoid arthritis and hence to improve its clinical effects.

Acute Disease ; Adolescent ; Adult ; Bridged-Ring Compounds ; poisoning ; Cardiomyopathies ; etiology ; therapy ; Creatine Kinase ; blood ; Creatine Kinase, MB Form ; Electrocardiography ; Foodborne Diseases ; blood ; complications ; therapy ; Glutamyl Aminopeptidase ; blood ; Humans ; Isoenzymes ; blood ; Male ; Middle Aged ; Nausea ; etiology ; Seizures ; etiology ; Treatment Outcome

OBJECTIVETo observe the effect of sodium tanshinone II (A) sulfonate (STS) on Ang II -induced atrial fibroblast collagen synthesis and TGF-beta1 activation.

METHODAtrial fibroblasts of neonatal rats were cultured to determine the content of collagen protein. The original synthesis rate determined by the [3H]-proline incorporation method was taken as the index for myocardial fibrosis. The content of active TGF-beta1 and total TGF-beta1 in cell culture supernatants were tested and cultured by ELISA. The expression of thrombospondin-1 (TSP-1) was assessed by using Western blot.

RESULTAng II could significantly increase the content of atrial fibroblast collagen and the collagen synthesis rate, the TSP-1 expression and the concentration of active TGF-beta1, without any obvious change in total TGF-beta1. After the STS treatment, all of the indexes, apart from total TGF-beta1, were obviously down-regulated.

CONCLUSIONSTS could decrease the secretion of Ang II -induced atrial fibroblast collagen and the synthesis rate. Its mechanism is related to the inhibition of TSP-1/TGF-beta1 pathway.

Angiotensin II ; pharmacology ; Animals ; Collagen ; biosynthesis ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Heart Atria ; cytology ; Phenanthrenes ; pharmacology ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Thrombospondin 1 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Objective To analyze the correlation of visual acuity and metamorphopsia with foveal morphology before and after vitrectomy with internal limiting membrane peeling (ILMP) in idiopathic macular epiretinal membrane (IMEM) eyes.Methods This is a retrospective case series of 47 IMEM patients (49 eyes).All the patients underwent 25G pars plana vitrectomy with IMEM removal and ILMP.The best corrected visual acuity (BCVA) was measured using the international standard visual acuity chart,and the results were converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity.The severity of metamorphopsia was measured using M-charts.The central macular thickness (CMT),inner nuclear layer thickness (INT),inner retinal layer thickness (IRT),outer retinal layer thickness (ORT),the status of ellipsoid zone (EZ) were assessed with spectral-domain optical coherence tomography at baseline and each month postoperatively.The differences in BCVA,CMT,INT,IRT,ORT and status of EZ before and after surgery were analyzed,so did the correlations between these indexes at the same time.Results Compared with baseline,the postoperative BCVA was significantly increased (F=6.133,P＜0.001),but the M value,CMT,INT,IRT,ORT were significantly decreased (F=12.481,10.565,15.739,6.046,10.569;P＜0.001);the integrity of EZ was improved significantly (x2=12.309,P＜0.001).Preoperative BCVA was positively related to the CMT (r=0.720) and ORT (r=0.720,0.887;P＜0.05),while negatively related to preoperative integrity of EZ (r=-0.295,P＜0.05).The postoperative BCVA was positively related to the preoperative BCVA and postoperative CMT,ORT (r=0.774,0.754,0.842;P＜0.05),while negatively related to postoperative integrity of EZ (r=-0.676,P＜0.05).The preoperative M value was positively related to the preoperative CMT,INT,IRT,and ORT (r=0.931,0.668,0.840,0.637;P＜ 0.05).The postoperative M value was positively related to the preoperative M value and postoperative CMT,INT,IRT,and ORT (r=0.723,0.722,0.767,0.825,0.387;P＜0.05).Conclusions Vitrectomy with ILMP for IMEM can improve the visual acuity,metamorphopsia and foveal morphology.Both visual acuity and metamorphopsia correlate with foveal morphology.

Resident standardization training is a major part of education of medical students after graduation , and is extremely important for high-level physician training and raising the quality of medical treatment. We trained the residents comprehensively according to the characteristics of echocardiography in terms of theoretical knowledge, clinical manipulative skills, clinical thinking ability, scientific research ability and communication and team cooperation ability, and carried out the relative examination according to the training. On the basis of practice, we explored the establishment of a standardized training mode and evaluation system suitable for echocardiography resident doctors in China.

Objective To evaluate the efficacy and safety of continuous suture by QuillTM SRS self-retaining suture for closing choledoch incision. Methods From January 2015 to October 2015, 66 patients with gall bladder calculi combined with common bile duct stones who received laparoscopic gallbladder excision, common bile duct explo-ration, laparoscopy choledochotomy, suture choledoch immediately were randomly divided into two groups, experi-mental group using QuillTM SRS self-retaining suture for closing choledoch incision and control group using tradi-tional absorbable suture for closing choledoch incision. Then observe the operation time, the risk of bile leakage oc-curred in the operation and the risk of bile leakage after operation. Results All patients successfully completed in laparoscopic surgery without laparotomy. Operation time, the risk of bile leakage occurred during operation in exper-imental group has statistically significant difference compared with control group ( < 0.05). The operation time and incidence of intraoperative bile leakage of the experimental group was less and lower than control group, while the risk of bile leakage after operation show no statistically difference ( >0.05). Conclusion Compared with interrupted suture by normal sutures, continuous suture by QuillTM SRS self-retaining suture for closing choledoch incision has the characteristics of shorter operation time and lower incidence of intraoperative and postoperative bile leakage. It is worthy of promoting.

The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Electrophoresis, Polyacrylamide Gel ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Proteomics ; Rats ; Rats, Sprague-Dawley