Objective To investigate sensitivity and specificity of myositis specific autoantibodies (MSAs) in polymyositis/dematomyositis(PM/DM ) and other neuromuscular disease. Methods Taking serum from 63 patients with PM/DM (PM/DM group)and 60 definite neuromusclar disease(non-myositis) patients(control group). All the sera were detected for two kind autoantibodies:anti-Jo-1,anti-SRP by immunoblotting method. Calculating sensitivity,specificity of anti-Jo-1 and anti-SRP autoantibodies for the diagnosis PM/DM. Results Positive ratio for anti-Jo-1 and anti-SRP autoantidodies were 17% and 5% respectively in the PM/DM group, while none of the sera from control group detected positive.The specificity of both autoantibodies diagnosis for PM/DM were 100% and (95% CI:94%~100%).The sensitivity was 22%( 95%CI:13%～34%). Conclusion Anti-Jo-1 and anti-SRP autoantibodies are highly specific to PM/DM diagnosis.

To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA (short hairpin RNA, or shRNA) was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein (EGFP) gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls (P>0.05), while the expression level in shRNA-transfected group decreased significantly (P<0.05). It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC12 cells.
Cloning, Molecular ; Gene Knockdown Techniques/*methods ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Myelin Proteins/*genetics ; Myelin Proteins/metabolism ; PC12 Cells ; Plasmids ; RNA Interference ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; RNA, Small Interfering ; Transfection

Objective　To evaluate the technique and clinical outcomes of modified transperitoneal laparoscopic radical prostatectomy.　Methods　A total of 285 patients received the operation with mean age of 67 years (50-76 years) from January 2008 to April 2012.Mean level of PSA was 15.7 μg/L (1.8 -50.0 μg/L),and mean prostatic volume was 44 ml (26 -74 ml). No lymph node or seminal vesicle involvement was found by CT or MR and radionuclide bone scan revealed no metastasis.271 cases were confirmed diagnosis by prostatic biopsy and 14 were detected through pathological studies of TURP specimens.Gleason score ranged from 6 to 8.14 cases were in clinical stage T1b,29 cases in T1c,214 cases in T2 and 28 cases in T3a.Transperitoneal approach and modified technique involving bladder neck dissection,nervesparing technique and vesicoureteral anastomosis were applied on patients.　Results　Mean operative time was 105 min (55 -150 min).Mean intraoperative estimated blood loss was 240 ml (50-800 ml).Rectal injures occurred in 2 cases and were repaired under laparoscopy.Drainage tube and urinary catheter were removed 48 -72 h and 5 -8 d postoperatively.Postoperative hospital stay was 7 d (5 - 11 d).Positive surgical margin was present in 58 patients.Mean follow-up time was 29 months (3 -50 months).Complete continence were found in 208 patients immediately after catheter removal.68 patient recovered continence within 3 months and 9 patients remained incontinence 3 months after surgery. Normal erection presented in 42 of the 57 cases with nerve-sparing.　Conclusions　Transperitoneal laparoscopic radical prostatectomy is safe and efficient.Higher efficiency and lower complication rate have been achieved through modified laparoscopic technique involving bladder neck dissection,nerve-sparing technique and vesicoureteral anastomosis.

To study the inhibitory effect of Nogo-A shRNA on cell line PC12, the Nogo-A shRNA (short hairpin RNA, or shRNA) was designed and synthesized. The annealed shRNA template was inserted into plasmid pGenesil-1 containing enhanced green fluorescent protein (EGFP) gene by gene cloning technique to generate eukaryotic expression vector. The recombinant plasmid was transfected into PC12 cells by lipofecamine2000 and the mRNA and protein expression level of Nogo-A gene was detected by RT-PCR and Western blotting 48 h after the transfection. Gene sequencing showed that that the Nogo-A shRNA eukaryotic expression vector was successfully constructed. No significant change was found in the Nogo-A mRNA and protein expression level in empty vector-transfected group as compared with controls (P＞0.05), while the expression level in shRNA-transfected group decreased significantly (P＜0.05). It is concluded that the pGenesil-1/Nogo-AshRNA recombinant plasmid can effectively suppress the expression of Nogo-A gene in PC 12 cells.

Studies on lycopene synthesis in Escherichia coli were not only able to gain the strains with high yield and less by-products, but also able to test functions of genes or gene clusters. In this article, the cDNA sequences of tomato LeGGPS2 and LePSY1 as well as the coding sequence of crtI from Erwinia uredovora, each of which was added a ribosome biding site, were controlled by T7 promoter and terminator alone or combined, and expressed in E. coli BL21 (DE3) to induce lycopene synthesis. The results show that only T7::crtI-LeGGPS2-LePSY1 expressed tri-cistronically could produce lycopene, and 2.124 mg/g dry cell weight oflycopene was obtained when fermented for 5 h at 30 degrees C after mixing 80 micromol/L IPTG at the later logarithmic phase while the seed broth of 1:50 (V/V) was inoculated into LB medium (pH 6.8) containing 3% sucrose and cultured for 8 h at 37 degrees C. The results confirmed the function of the prokaryonized LeGGPS2 and LePSY1 and their synergy with crtI, and also laid a foundation to establish an independent lycopene synthetic pathway in the tomato plastid.
Bacterial Proteins ; genetics ; Carotenoids ; biosynthesis ; genetics ; Cloning, Molecular ; Erwinia ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Engineering ; Plant Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVE:To explore new method of prescription comment in medical institutions of Chongqing under the background of"New Medical Reform",and to provide reference for prescription comment innovation and promoting rational drug use in clinic. METHODS:Through analyzing policy background of prescription comment innovation and practice,referring to experience of other regions,the achievements and problems were analyzed by developing centralized prescription comment and prescription comment information construction in municipal and regional (county-level) medical institutions of Chongqing. The countermeasures suggestion was put forward. RESULTS&CONCLUSIONS:The prescriptions of 16 municipal medical institutions were commented centrally,involving more than 300 medical records and 1600 prescriptions or medical orders. The prescriptions of 135 regional (county-level) medical institutions were commented centrally,involving more than 3100 medical records and 12100 prescriptions or medical orders."Chongqing Medical Institution Prescription Comment Key Technology Monitoring System"was established. Above measures innovated centralized prescription comment mode,improved the quality of prescription comment and promoted prescription comment informatization. There were still problems as incomplete coverage of the system,inspection to become a mere formality. It is suggested to strengthen internal quality control of medical institutions by establishing rational drug use scoring management system,developing clinical pharmacist ward round and prescription checking in advance;establish and improve third-party external comment and supervision mechanism;improve the knowledge of prescriptions comment staff in all directions so as to further standardize prescriptions and promote rational drug use in medical institutions.

Objective:To investigate the therapeutic effect of skull drilling and/or grinding combined with artificial dermis and vacuum sealing drainage in repairing scalp defects with skull exposure.Methods:From October 2014 to May 2018, 18 patients with scalp defect and skull exposure were treated in the Department of Burn and Plastic Surgery, the Second Clinical College of North Sichuan Medical College, including 10 males and 8 females, with an average age of 64 years (range, 34-86 years). The patients were divided into two groups: group A (by drilling skull or/and grinding combined with artificial dermis cover and vacuum sealing drainage plus two split thickness skin graft repair) and group B (by drilling skull or/andgrinding combined with artificial dermis cover plus two covering leather grinding stage split thickness skin graft repair), 9 cases in each group. The head wound granulation tissue, postoperative complications, skin graft survival rate and wound healing time were compared between the two groups. Vancouver scar assessment scale (VSS) was used to evaluate the wound healing in the two groups.Results:The time of granulation cultivation in group A and group B was (16.44±1.42) days and (29.11±13.32) days, the difference was statistically significant ( P<0.05); The wound healing time of group A and group B was (26.00±3.32) days and (40.67±14.37) days, the difference was statistically significant ( P<0.05); The postoperative complications of group A and group B were 1 case and 5 cases respectively, the difference was statistically significant ( P<0.05). The skin graft survival rates of group A and group B were (97.11±3.44)% and (95.00±4.74)%, the difference was not statistically significant ( P>0.05); The wound scar VSS scores of group A and group B were (7.67±1.32) points and (8.78±1.99) points, the difference was not statistically significant ( P>0.05). Conclusions:By drilling skull and/or grinding combined with artificial dermis cover and vacuum sealing drainage and two stage split thickness skin graft for repairing scalp defect with skull exposure wound can not only better scalp defect with skull exposure wounds, and reduce the postoperative complications, and significantly accelerate wound healing, but also can effectively improve the quality of wound healing, which is worthy of clinical application.

ObjectiveTo investigate the clinical features of patients with hepatolenticular degeneration （HLD） aged above 35 years. MethodsA retrospective analysis was performed for the clinical data of the patients with HLD， aged above 35 years， who attended West China School of Public Health， Sichuan University， from April 2018 to April 2023， and according to their clinical symptoms， they were divided into mixed type group with 13 patients， liver type group with 12 patients， and brain type group with 5 patients. Related data were collected， including general information （sex， clinical manifestation， age at confirmed diagnosis， time from initial symptoms to confirmed diagnosis， and family history）， laboratory examination （routine blood test， liver and renal function， serum copper， serum ceruloplasmin， urinary copper， and coagulation function）， and radiological examination. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups； the Fisher’s exact test was used for comparison of categorical data between groups. ResultsFor the 30 patients with HLD， the male/female ratio was 3∶1， and the mean age was 46.13±5.88 years； the patients with positive Kayser-Fleischer ring of the cornea accounted for 43.33%， and the patients with liver cirrhosis accounted for 66.67%. There were significant differences between the three groups in globulin， albumin/globulin ratio， alanine aminotransferase， prothrombin time， international normalized ratio， and activated partial thromboplastin time （F=5.893， 4.513， 4.424， 5.029， 5.248， and 4.942， all P<0.05）. ConclusionMost patients are male among the patients diagnosed with HLD after 35 years of age， with the main clinical types of mixed type and liver type， and such patients tend to have poor liver and coagulation functions. For unexplained liver function abnormalities and liver cirrhosis in this age group， the indicators such as serum ceruloplasmin and urinary copper should be screened as early as possible， and liver and kidney function and coagulation function should be monitored.

OBJECTIVETo study the technique and clinical outcomes of laparoscopic radical prostatectomy for high risk prostate cancer.

METHODSA total of 65 patients with high risk prostate cancer were treated with surgery in the First Affiliated Hospital of Nanjing Medical University from January 2011 to June 2013. The mean age was 67 years (range 45-75 years). The mean preoperative prostate specific antigen (PSA) level was 26.7 µg/L (range 11.2-65.5 µg/L). The transrectal biopsy revealed Gleason score of 3+3 in 4 patients, Gleason 3+4 in 27 patients, Gleason 4+3 in 11 patients, Gleason 4+4 in 21 patients and Gleason 4+5 in 2 patients. The bone metastasis was excluded by scintigraphy examination. The surgical procedures were performed through transperitoneal approach. Extended pelvic lymph nodes dissection was performed after the removal of the prostate. Adjuvant radiotherapy or hormonal therapy was administrated according to the pathological results. Serum PSA was detected every 1 to 2 month and urinary continence was evaluated every 3 month in the first year, and then serum PSA was detected every 2 to 3 month.

RESULTSThe mean operative time was (134±21) minutes and the median blood loss was (300±146) ml. Bladder neck reconstruction was performed in 15 cases. The drainage was removed on postoperative day 4 and the catheter was removed on day 7. Pathologic results demonstrated pT2 in 25 patients, pT3a in 28 patients, pT3b in 9 patients and pT4 in 3 patients. Positive surgical margin was presented in 15 patients. A median of 19 lymph nodes (range 11-24 nodes) were retrieved during lymphadenectomy and 11 patients had lymph nodes metastasis with a total of 19 positive nodes. Forty-three patients recovered continence after the removal of catheter. Eleven patients received adjuvant hormonal therapy and 19 patients received adjuvant radiation therapy. With the median of 20 months follow-up (range 12-30 months), 5 patients got biochemical recurrence.

CONCLUSIONSLaparoscopic radical prostatectomy with extended lymph nodes dissection for high risk prostate cancer is safe and technical feasible. It provides accurate information on tumor stage and grade. It is an important component of multimodality for the treatment of high risk prostate cancer.

Aged ; Biopsy ; Humans ; Laparoscopy ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; Postoperative Period ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; diagnosis ; surgery