1.Clinical detection and identification of seven species human herpesviruses by DNA microarray technique
Yanmin ZHU ; Yidong WU ; Shiqiang SHANG
Chinese Journal of Clinical Laboratory Science 2006;0(05):-
Objective To establish a method for rapid diagnosis and identification of seven species human herpesviruses infection.Methods Primers and oligonucleotide probes were designed and synthesized based on the highly conserved regions of the DNA polymerase genes in human herpesviruses, namely herpes simplex virus type 1 (HSV-1),HSV-2,varicella-zoster virus (VZV),Epstein-Barr virus (EBV),cytomegalovirus (CMV),and human herpes virus 6 (HHV-6A/6B).DNA microarrays were made by printing the oligonucleotide probes on the special glass slides.A total of 282 blood specimens from children with suspected infection were analyzed by this DNA microarray technique,and the results were compared with those of TaqMan PCR.Results The products of the seven human herpesviruses after PCR amplification could be used to identify the virus species with DNA microarrays.The detection limits were 10 copies/?l for HSV-1,HSV-2,VZV,EBV,CMV,HHV-6A,and HHV-6B,respectively.The assay did not show cross-reaction to the DNA extract of hepatitis B virus,staphylococcus aureus,E.coli,Candia albicans and human genome.Among the 282 samples,59 were positive for human herpesviruses DNA.Compared with those of TaqMan PCR,the sensitivity and specificity of the microarray assay were 96.7% and 99.5%,respectively,and the index of accurate diagnosis was 0.962.Conclusions This DNA microarray for identifying human herpesviruses species is specific and sensitive,and may serve as an efficient technique for simultaneous detection and species identification of human herpesviruses in clinical specimen.
2.An experimental study on repairing sciatic nerve defects of rats by human umbilical cord derived mesenchymal stem cells and acellular nerve basal lamina tube
Xuejian WU ; Jiangtao HE ; Shiqiang SUN
Chinese Journal of Microsurgery 2010;33(6):461-464,后插7
Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.
3.Using of 16S rRNA gene chip hybridization in the diagnosis of neonatal sepsis
Jiyan ZHENG ; Shiqiang SHANG ; Yidong WU
Chinese Journal of Infectious Diseases 2001;0(03):-
Objective To improve the speed and accuracy of bacteria detection, and develop the test of 16S rRNA genes PCR amplification plus gene chip hybridization to diagnose neonatal sepsis. Methods Bacterial 16S rRNA genes were detected in blood and CSF samples of 125 suspected neonatal sepsis, and the results were compared with blood culture, CSF culture, and non-specific diagnostic parameters (WBC, PLT, CRP). 30 non-infectious neonates were regarded as the negative control group. Gene chip test were performed by extraction of DNA, primers and probes design, PCR amplification, preparation of gene chip, hybridization, laser scan and reading of the results. 18 specific probes, including universal 1, universal 2, Gram positive bacterial probe, Gram negative bacterial probe 1, Gram negative bacterial probe 2, Staphylococcus aureus, Staphylococcus epidermidis, CoNS (Coagulase Negative Staphylococcus), Escherichia coli, Haemophilus influenzae, Listeria monocytogenes, Streptococcus pneumoniae, Streptococcus agalactiae, Bacteroides fragilis, Bacillus, Meningococcus, Corynebacterium, Propionibacterium, were used in the test. Results The positive rate of PCR test was 51.2% in 125 blood samples, and was significantly higher than the positive blood culture (25.6%), or the indicator of two abnormal non-specific parameters (32.8%) (P
4.Real-time quantitative PCR in detection of neonatal sepsis
Rongshan WANG ; Yidong WU ; Shiqiang SHANG
Chinese Journal of Perinatal Medicine 2000;0(04):-
Objective To explore a rapid diagnostic method in neonatal sepsis and bacterial me- ningitis. Methods The primers and TaqMan probes were designed and synthesized based on the sequences of bacterial 16S rRNA gene. Nineteen bacterial strains, 3 different viruses, fungus and human genomic DNA were tested by FQ-PCR assay. Blood specimens and CSF from 195 cases of suspected septicemia were detected with both TaqMan PCR assay and blood or/and CSF culture. Results The FQ-PCR showed very high sensitivity and specificity and was able to detect at least 10 copies of 16S rRNA gene equivalent to 1~2 copies bacterium. No cross-reaction was found with human genomic DNA, other fungus and viruses. Among the 195 cases, the positive rate by FQ-PCR was 12.8%(25 cases) and 7.1%(15 cases) by blood culture ( P
5.Effect of UK combined with tirofiban in young patients with ST segement elevation acute myocardial infarction
Gaizhen ZHU ; Ling LI ; Shiqiang WEI ; Huijun LIU ; Xinsheng WU
Chinese Journal of Primary Medicine and Pharmacy 2015;(10):1444-1446,1447
Objective To study the thrombolysis effect and safety of UK combined with tirofiban in young patients with ST segement elevation acute myocardial infarction.Methods 76 young patients with ST segement elva-tion acute myocardial infarction were selected as research objects,and they were divided into UK group(control group, n =34)and UK combined with tirofiban group(observation group,n =42)by random number table,then the recanali-zation rate of infarct -related artery and incidence of complications were compared.Results The recanalization rate of infarct -related artery in the observation group was 97.6%,which was 91.2% in the control group,there was no significant difference between the two groups(χ2 =1.564,P >0.05);The vascular infarction related to blood flowⅡlevel of the patients was 90.5% in the observation group,which was significantly higher than 70.6% in the control group(χ2 =4.945,P <0.05);The death,complications incidences had not significant differences between the two groups(χ2 =1.252,2.837,2.837,all P >0.05).Conclusion The thrombolysis effect of UK has a high recanaliza-tion rate of infarct -related artery in young patients with ST segement elevation acute myocardial infarction,and the thrombolysis effect of UK combined with tirofiban is better,and it does not increase the incidence of complications.
6.A retrospective clinical study of urogenital fistula caused by gynecological and obstetrical surgery
Jianhui WU ; Shiqiang YANG ; Yong XU ; Hongshun MA
Chinese Journal of Urology 2014;35(9):686-690
Objective To study the etiology,diagnosis,treatment and prophylaxis strategy of the urogenital fistula caused by gynecological and obstetrical surgery.Methods Data of 64 cases with urogenital fistula,who were admitted into second hospital of Tianjin medical university and Tianjin first central hospital from January 1992 to December 2012,were analyzed retrospectively.In Tianjin first central hospital,those cases include vesicovaginal fistula in 10,ureterovaginal fistula in 7 and urethro-vaginal fistula in one case.In second hospital of Tianjin medical university,those cases include vesicovaginal fistula in 26,ureterovaginal fistula in 18,urethro-vaginal fistula in 1 and ureterouterine fistula in one case.The median age was 42 years old (range 21-53).The history of diseased ranged from 16 days to 30 years.All patients were diagnosed by methylene blue test,cystoscopy,ureteroscopy,intravenous urography,ultrasound,computed tomography (CT) and magnetic resonance urography (MRU).The primary fistula was diagnosed in 50 cases and the recurrence was found in 14 cases.Single fistula existed in 56 cases and multiple fistulas were found in 8 cases.In 36 patients with vesicovaginal fistula,transabdominal repair of vesicovaginal fistula (n =20),transpubic surgery (n=10) and transvaginal surgery (n=6) were chosen.In 25 patients with ureterovaginal fistula,ureterocystostomy (n =10),ureterotomy with holmium laser (n =8),ureteral stent placement (n =6) and ureteral stricture excision and bladder-psoas suspension (n=1) were used.Two patients with urethro-vaginal fistulae were cured by the Latzko technique.One patient had uretero-uterine fistula and cured by ureteral stricture excision,ureterocystostomy and bladder-psoas suspension.Results Fifty-five(86%) cases were cured by single-stage surgical treatment and nine patients experienced more than two times of surgical treatment.The incipient patients have a higher success rate of first surgery than recurrent patients (92% vs.64%,P<0.05).Single and multiple fistulas have no significant difference about the surgical successive rate (88% vs.75%,P>0.05).In cases with vesicovaginal fistula,the success rate of vaginal and abdominal approaches are the same 85% (P>0.05).In cases with ureterovaginal fistula,abdominal and endoscopic approaches were 100% and 85%,respectively (P>0.05).The mean duration of follow was 20 months (range 3-48).There was no recurrence during follow-up.Conclusions Urogenital fistula caused by gynecological and obstetrical operation can be cured by surgery.Recurrent fistula is a challenge for diagnose and treatment,preoperative need reasonable operation mode to improve the success rate of operation.Both open surgery and endourology approaches are effective treatment options in management the urogenital fistula.
7.Simultaneous HPLC determination of 9 effective components ofTanreqing injection
Yonggang CHEN ; Shiqiang WU ; Chuanxue MA ; Bing HU
International Journal of Traditional Chinese Medicine 2015;37(9):817-821
Objective To establish HPLC method for the simultaneous detgrmination of 9 effective components ofTanreqing injection.Methods The Agilent TC-C18 (2) (250 mm×4.6 mm, 5μm) column was used, the mobile phase consisted of acetonitrile (A) acetonitrile-0.1% formic acid aqueous solution (B) in a gradient mode, the flow rate was 1.0 ml/min,the column temperature was 30℃, the injection volume was 10μl, and the detecting wavelength was at 280 nm.Results All the 9 effective components showed a good linear relationship. TheRSDof the precision, reproducibility and stability tests were less than 2%. The average recoveries of the 9 effective components were in the range of 99.3%-103.1%.Conclusion This analysis method is simple, accurate and has a good specificity, which can be suitable for controlling the quality of Tanreqing injection.
8.THE NUTRITIONAL VALUE AND THE EDIBLE SAFETY OF FASTFOOD EUCHEUMA
Liudong LI ; Peiji CHEN ; Xianqing YANG ; Yanyan WU ; Shiqiang DIAO ; Laihao LI
Acta Nutrimenta Sinica 1956;0(04):-
Objective:The nutritional value and edible safety were evaluated for fastfood Eucheuma. Methods:The nutritional composition and the contents of heavy metal element and microbiology index were tested, and the test of acute toxicity and mutation were performed to evaluate the nutritional value and edible safety. Results:The main components of fastfood Eucheuma were dietary fibre and mineral components, and the contents of protein and fat were very low. The gross content of dietary fibre was 88.6%. The contents of Zn and Ca were the highest among the mineral components.The index of heavy metal elements and microbes(cfu) were lower than that of the national standard. It was non-toxic and non-mutagenic. Conclusion:The fastfood Eucheuma is a functional food, and also a safe food of new resource.
9.Application of three-dimensional reconstruction using Mimics software to repair of Pilon fracture
Jian HUANG ; Xiaoping WANG ; Zhicheng DENG ; Weiwei WU ; Luyao CHEN ; Shiqiang HU ; Zhantu WEI ; Sheng GUO
Chinese Journal of Tissue Engineering Research 2015;(44):7167-7171
BACKGROUND:Mimics software is a three-dimensional (3D) image processing and editing tool based on CT scan data. Mimics software can rebuild the data and images gotten in CT, MRI and ultrasound scans into 3D images and display on the computer screen so as to help clinicians understand the type of fracture and the relationship of the 3D structure of the surrounding tissue and to provide a great help in the development of orthopedics operation. OBJECTIVE:To explore the application effect of Mimics software 3D reconstruction on perioperative period of Pilon fracture. METHODS:This study selected 61 cases of Pilon fracture, who received the surgery in the Zhongshan City Xiaolan People’s Hospital from September 2008 to September 2013, as research objects. They were randomly divided into 3D group and control group in accordance with the time of admission. Al patients underwent anterioposterior and lateral X-ray film examination and multi-slice spiral CT scan. Patients in the control group received internal fixation according to above examination results. Patients of the 3D group, on the base of those of the control group, were subjected to internal fixation after three-dimensional entity reconstruction by using Mimics V 10.0 software. Operation time and functional curative effect in the postoperative folow-up were compared between the two groups. RESULTS AND CONCLUSION:The operation time was significantly less in the 3D group than in the control group (P < 0.05). The number of patients with excelent 6-month functional curative effect was more, and the number of patients with poor effect was less in the 3D group than in the control group. The 6-month functional curative effect was better in the 3D group than in the control group (P < 0.05). These results showed that the application of Mimics software 3D reconstruction to the perioperative period of Pilon fractures can give comprehensive assessment of the situation of fracture, optimize and improve the preoperative plan and reduce the risk of surgery, and promote the successful completion of internal fixation.
10.High performance liquid chromatography analysis of porcine platelet-derived gro wth factor and its effect on DNA synthesis of human vascular endothelial cells
Yusheng REN ; Zonggui WU ; Fang CUI ; Guoliang JIA ; Shiqiang YU ; Qiang CHEN
Academic Journal of Second Military Medical University 2001;22(2):155-158
Objective: To determine the molecular weight and p urity of porcine platelet-derived growth factor (pPDGF) and to investigate its effect on DNA synthesis of human umbilical vein endothelial cells. Metho ds: In the present experiment, the high performance liquid chromatograph y was used and the molecular weight and purity of pPDGF were studied. Human umbi lical vein endothelial cells was cultured and effects of pPDGF on DNA synthesis of endothelial cells was observed by 3H-TdR incorporation in vitro. Results: The findings of high performance liquid chromatography showed that the molecular weight of pPDGF was 29 120 and the purity was 89.46%, a nd pPDGF significantly promoted DNA synthesis of quiescent endothelial cells wit h a maximal response at a concentration of 40 ng/ml at 48 h. Conclusion: The molecular weight of pPDGF is 29 120, and it can promote DNA synthes is of cultured human umbilical vein endothelial cells.