In order to study the relationship between lengths of homologous fragments and chromsomic recombination rate in Saccharopolyspora erythraea, three homologous sequences, with mutant loci and different flanking sequences, (26bp + 27bp), (500bp + 576bp) and (1908bp + 1749bp), were synthesized by chemical reaction or PCR amplification, and cloned into pWHM3 to construct homologous recombination plasmids, pWHM1113, pWHM1116 and pWHM1119. When the plasmids were transformed into protoplast of Saccharopolyspora erythraea A226 under PEG mediated, on an average 30, 69 and 170 transformants grew on each plate for the three plasmids respectively, but chromosomic integration frequency were 0, 2% and 19% among corresponding transformants. Both pWHM1116 and pWHM1119 could take double crossover recombination, and exchange the mutant loci in the chromosome. It was concluded that when the flanking sequences were equal or more than (500bp + 576bp), they could take effective single and double recombination with Saccharopolyspora erythraea chromosome.
Chromosomes, Bacterial ; genetics ; Polymerase Chain Reaction ; Recombination, Genetic ; genetics ; Saccharopolyspora ; genetics

Adult ; Aged ; Biomarkers, Tumor ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; metabolism ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Liver Cirrhosis ; metabolism ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Retinoblastoma Protein ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo analyze the characteristics of CDR3 of TCRbeta on CD8+ T cells in chronic hepatitis B patients.

METHODSEight patients with chronic hepatitis B (ALT more than 2 ULN) were enrolled in this study. CD8+ T cells were isolated from peripheral blood. RT-PCR was proformed to amplify the CDR3 of TCRbeta, and the PCR products were sequenced and analyzed.

RESULTSThe chronic hepatitis B patients showed obvious clonal expansion of T cell, and three perturbation patterns of T cell expansion were showed in the CDR3 of TCRbeta, including monoclonicity, oligoclonicity and skewed peak patterns. The number of perturbation families of CD8+ subpopulation was significantly higher than that of CD8- subpopulation (10.6+/-4.7 vs. 4.1+/-3.1, t = 6.619, P less than 0.01). In 3 out of 8 patients, the number of perturbation families of CD8+ subpopulation was also higher than that of PBMCs without depleting CD8+ subpopulation.

CONCLUSIONSThe characteristics of CDR3 of TCRbeta may help to understand the inflammatory response in CHB patients.

Adult ; CD8-Positive T-Lymphocytes ; immunology ; Complementarity Determining Regions ; genetics ; Genes, T-Cell Receptor beta ; Hepatitis B, Chronic ; blood ; immunology ; Humans ; Male ; Receptors, Antigen, T-Cell ; immunology ; Young Adult

Adolescent ; Adult ; Aged ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Splenectomy ; methods

R-spondin2 (Rspo2) is a member of protein family RSPOs, which can be coupled to receptor 4/5 (leucine-rich repeat-containing g protein-coupled receptor 4/5, LGR4/5), cell surface transmembrane E3 ubiquitin ligase ZNRF3/RNF43 (zinc and ring finger 3/ring finger protein 43), heparan sulfate proteoglycan (heparan sulfate proteoglycans, HSPGs) and the IQ motif (IQ gap 1) containing GTP enzyme activating protein 1, regulating the Wnt/β-catenin signaling pathway, which is the most widely studied signaling pathway and directly related to basic bone biology. Any problem in this pathway may have an impact on bone regulation. In recent years, it has been found that Rspo2 can act on osteoblast, osteoclast and chondrocytes through Wnt/β-catenin, and take part in occureace and development of some bone diseases such as ossification of the posterior longitudinal ligament (OPLL), osteoarthritis (OA) and rheumatoid arthritis (RA), so the study of Rspo2 may become a new therapeutic direction for bone-related diseases. Based on the latest research progress, this paper reviews the structure and main functions of Rspo2, the mechanism of Rspo2 regulating Wnt/β-catenin signaling pathway and its influence on skeletal system, in order to provide new ideas and ways for the prevention and treatment of bone-related diseases.

BACKGROUNDElectroconvulsive therapy (ECT) can alleviate the symptoms of treatment-resistant depression (TRD). Functional network connectivity (FNC) is a newly developed method to investigate the brain's functional connectivity patterns. The first aim of this study was to investigate FNC alterations between TRD patients and healthy controls. The second aim was to explore the relationship between the ECT treatment response and pre-ECT treatment FNC alterations in individual TRD patients.

METHODSThis study included 82 TRD patients and 41 controls. Patients were screened at baseline and after 2 weeks of treatment with a combination of ECT and antidepressants. Group information guided-independent component analysis (GIG-ICA) was used to compute subject-specific functional networks (FNs). Grassmann manifold and step-wise forward component selection using support vector machines were adopted to perform the FNC measure and extract the functional networks' connectivity patterns (FCP). Pearson's correlation analysis was used to calculate the correlations between the FCP and ECT response.

RESULTSA total of 82 TRD patients in the ECT group were successfully treated. On an average, 8.50 ± 2.00 ECT sessions were conducted. After ECT treatment, only 42 TRD patients had an improved response to ECT (the Hamilton scores reduction rate was more than 50%), response rate 51%. 8 FNs (anterior and posterior default mode network, bilateral frontoparietal network, audio network, visual network, dorsal attention network, and sensorimotor network) were obtained using GIG-ICA. We did not found that FCPs were significantly different between TRD patients and healthy controls. Moreover, the baseline FCP was unrelated to the ECT treatment response.

CONCLUSIONSThe FNC was not significantly different between the TRD patients and healthy controls, and the baseline FCP was unrelated to the ECT treatment response. These findings will necessitate that we modify the experimental scheme to explore the mechanisms underlying ECT's effects on depression and explore the specific predictors of the effects of ECT based on the pre-ECT treatment magnetic resonance imaging.

Adult ; Brain ; pathology ; physiopathology ; Depression ; physiopathology ; therapy ; Depressive Disorder, Treatment-Resistant ; physiopathology ; therapy ; Electroconvulsive Therapy ; methods ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged

OBJECTIVE: To compare the therapeutic effect between thunder-fire moxibustion combined with external applicaion of powder and thunder-fire moxibustion alone for mild and moderate knee osteoarthritis. METHODS: A total of 70 patients with mild and moderate knee osteoarthritis were randomly divided into an observation group and a control group, 35 cases in each group. In the observation group, thunder-fire moxibustion combined with external applicaion of powder were applied, Thunder-fire moxibustion was applied at Xuehai (SP 10), Liangqiu (ST 34), Neixiyan (EX-LE 4), Yanglingquan (GB 34) and point, external applicaion of powder was given to the affected knee after the treatment of thunder-fire moxibustion. Simple thunder-fire moxibustion was given in the control group. All patients in the two groups were treated once a day, 7 days as one course and the consecutive 4 courses were required, with an interval of 1 day between courses. Before and after treatment, the visual analogue scale (VAS) score and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score were used to assessed knee pain, stiffness and physical function in the two groups. In addition, the efficacy was evaluated. RESULTS: Compared before treatment, the VAS scores, total scores of WOMAC, pain scores, stiffness scores and physical function scores were reduced after treatment in the two groups (<0.01), and the scores in the observation group were significantly lower than those in the control group (<0.01, <0.05). The total effective rate was 97.0% (32/33) in the observation group, which was superior to 91.2% (31/34) in the control group (<0.05). CONCLUSION Thunder-fire moxibustion combined with external applicaion of powder are superior to simple thunder-fire moxibustion in improving the symptoms and delaying the development of the disease for mild and moderate knee osteoarthritis.
Acupuncture Points ; Humans ; Knee Joint ; Moxibustion ; Osteoarthritis, Knee ; therapy ; Treatment Outcome

AIM: To evaluate the efficacy of modified folding intraocular lens(IOL)suspension surgery in treatment of traumatic dislocation of lens surgery technique.METHODS: Prospective randomized controlled study. A total of 15 patients underwent the modified folding IOL suspension surgery. Among them, 9 patients chose Akreos AO IOL, and polypropylene sutures were used to thread the haptics of IOL. After guided to puncture out through the sclera, the ends of sutures were thermal expanded and fixed in the sclera. And 6 patients chose Tecnis ZA9003 IOL and no sutures were used. After guided the haptics to puncture out through the sclera, the ends of haptics were thermal expanded and fixed in the sclera. The best corrected visual acuity(BCVA, LogMAR)of all patients and postoperative complication were observed. RESULTS: This study included 15 patients, among them, 7 were male and 8 were female, the mean age was 64.00±9.85 years old, the mean course of diseases was 5.80±3.17 wk. There were no significant differences between the demographic and baseline clinical characteristics. After underwent the modified folding IOL suspension surgery, visual acuity of all patients were obviously improved. After 3mo of the surgery, the BCVA(LogMAR)of patients were improved from 1.28±0.56 to 0.52±0.30. More specifically, the BCVA(LogMAR)of patients who chose Akreos AO IOL were improved from 1.39±0.62 to 0.59±0.25, and those who chose Tecnis ZA9003 IOL of the BCVA(LogMAR)were improved from 1.12±0.45 to 0.42±0.35. Furthermore, there was no severe postoperative complication observed in our study. Only one patient suffered IOL dislocation and the IOL optical surface was mild oblique.CONCLUSION: Modified folding IOL suspension surgery technique resulted in good visual and outcomes with no severe complication, making it an effective option for IOL suspension surgery.

Objective: To evaluate the safety and long-term clinical efficacy of percutaneous coronary intervention (PCI) in patients with in-stent chronic total occlusion (IS-CTO) lesions. Metheds: This is a retrospective analysis. Patients with IS-CTO who underwent PCI in Fuwai hospital from January 2010 to December 2013 were enrolled. A total of 212 patients who met the inclusion criteria were included in the IS-CTO group, 212 matched patients with primary CTO lesions were included in the de novo CTO group. The incidence of complications and the success rate of PCI were compared between the two groups. Successful PCI was defined as successfully implantation of stent(s) at target CTO lesions. The primary endpoint was defined as a composite event of cardiac death and myocardial infarction (MI). Secondary endpoints including PCI success, all-cause death, cardiac death, MI, target vessel related MI, revascularization, target vessel revascularization, heart failure for rehospitalization. The patients were followed up for 5 years after PCI. Results: A total of 424 cases were included. The mean age was (57.8±10.5) years, there were 364 males in this cohort. The left ventricular ejection fraction was significantly lower ((58.7±9.2)% vs. (61.0±7.7)%, P=0.01) and the SYNTAX scores was significantly higher (19.4±8.3 vs. 15.3±10.0, P<0.01) in IS-CTO group than that in de novo CTO group. The proportion of patients with target CTO lesions in left anterior descending artery was significantly higher (42.9% (50/212) vs. 23.6% (91/212), P<0.01) in IS-CTO group than that in de novo CTO group. The rate of successful PCI (71.7% (152/212) vs. 69.8% (148/212), P=0.70) and complication (40.6% (86/212) vs. 36.3% (77/212), P=0.37) was similar between the two groups. The incidence of primary endpoint at 5 years was significantly higher in IS-CTO group (10.8% (23/212) vs. 4.7% (10/212), P=0.02), which was driven by higher incidence of MI (9.0% (19/212) vs. 4.2% (9/212), P=0.05). There were a trend of higher secondary endpoints in IS-CTO group (all P>0.05). Conclusion: The safety and effectiveness of PCI are acceptable in patients with IS-CTO, but the risk of long-term cardiac death and MI is higher among patients with IS-CTO as compared to patients with primary CTO lesions.