2.Construction of a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein
Zhenghao TANG ; Huihui MA ; Gang LI
Chinese Journal of Infectious Diseases 2001;0(06):-
Objective To construcr a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein. Methods The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by RT-PCR from peripheral blood Lymphocytes mRNA of patients with positive serum hepatitis B virus core protein antibody and randomly combined through a DNA linker encoding the peptide (Gly 4Ser) 3 to construct single-chain variable fragments (ScFv)gene. In the presence of helper phage M13KO7, the ScFv were displayed on the surface of recombinant phages. Results Repertoire antibody library was obtained with 10 6 clonies resistant to ampicillin and kanamycin. Conclusions A typical phage display library of human single-chain Fv antibodies has been constructed Using RT-polymerase chain reaction (RT-PCR) and phage display technique, human single-chain Fv antibodies can be screened from this library.
3.Energy calibration and electrical system design of HK.ESWL-008 child extracorporeal shock wave lithotripter
Shihong MA ; Guangqing LI ; Shuai YAN ; Gang LI ; Ming CHEN
Chinese Medical Equipment Journal 2017;38(6):46-50
Objective To design an electrical system of extracorporeal shock wave lithotripter for children and adults uses.Methods The shock wave kernel parameters of child energy section of Dornier Delta Ⅱ lithotripter were tested such as peak sound pressure,pressure pulse width and pressure pulse rise time,and then energy calibration was executed for HK.ESWL008 child extracorporeal shock wave lithotripter to make the kernel parameters of the two lithotripters the same or similar to each other.An electrical system was designed with PLC system as the core control unit,involving in wave source 3D motion,large and small C-arms motion,water circulation system,B-type ultrasound positioning system,high-frequency X-ray imaging system,high-voltage condenser charging and discharging system,console and bedside-box-controlled display system as well as electromagnetic compatibility.Results Within A,B,C,1 and 2 energy sections for child lithotripsy the maximum and average differences between the kernel parameters of the two lithotripters were 7.1% and 3.8% respectively.There was no difference between the clinical experience by the two lithotripters,and HK.ESWL-008 child extracorporeal shock wave lithotripter passed EMC and safety detections by Guangdong Medical Devices Quality Surveillance and Test Institute of CFDA.Conclusion HK.ESWL-008 child extracorporeal shock wave lithotripter can be used for children lithotripsy clinically.
4.Comparison study on lung function impairment degree between stage II+ and III.
Gang CHEN ; Guo-Xuan MA ; Yan-Jun LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(1):47-48
Adult
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Aged
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Humans
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Male
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Middle Aged
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Pneumoconiosis
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pathology
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physiopathology
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Respiratory Function Tests
5.Clinical Observation of Levofloxacin and Capreomycin Combined with Chemotherapy Regimen in the Treat-ment of Multi-drug Resistant Tuberculosis
Ming LIU ; Yinglian MA ; Yonggui GANG ; Quanlu ZHANG ; Weihong LI
China Pharmacy 2016;27(27):3788-3790
OBJECTIVE:To observe the efficacy and safety of levofloxacin and capreomycin combined with chemotherapy regi-men in the treatment of multi-drug resistant tuberculosis(MDR-TB). METHODS:84 MDR-TB patients were randomly divided in-to observation group (42 cases) and control group (42 cases). Observation group received 0.75 g Capreomycin sulfate for injec-tion,addint into 100 ml 0.9% Sodium chloride injection,intravenous infusion,once a day+0.4 g Levofloxacin hydrochloride tab-let,orally,once a day+0.2 g Protionamide tablet,orally,3 times a day+0.3 g Pasiniazid tablet,orally,3 times a day+0.5 g Pyra-zinamide tablet,orally,4 times a day. Control group received 0.4 g Amikacin sulfate injection,adding into 100 ml 0.9% Sodium chloride injection,once a day,intravenous infusion+0.3 g Ofloxacin tablet,orally,twice a day+Protionamide tablet (the same dose with observation group)+Pasiniazid tablet (the same dose with observation group)+Pyrazinamide tablet (the same dose with observation group). All patient were given 0.1 g Glucuronolactone tablet,orally,3 times a day. The treatment course for both group was 12 months. Sputum negative conversion rate,negative conversion time,symptom improvement time,lesion absorption and lung cavity closing,and cell immune indexes (CD4+CD25+/CD4+,CD4+CD25+CD127low/CD4+),IL-17 level before and after treatment,and the incidence of adverse reactions in 2 groups were observed. RESULTS:The sputum negative conversion rate,ab-sorption rate,lung cavity closing and narrowing cases in research group after 3,6,9,12,18 months treatment were significantly higher than control group,sputum negative conversion time,symptom improvement time in observation group were significantly lower than control group,the differences were statistically significant(P<0.05). Before treatment,there were no significant differ-ences in CD4+CD25+/CD4+,CD4+CD25+CD127low/CD4+,IL-17 level in 2 groups(P>0.05). After treatment,CD4+CD25+/CD4+,CD4+CD25+CD127low/CD4+ in 2 groups were significantly lower than before,and observation group was lower than control group,IL-17 level was significantly higher than before,and observation group was higher than control group,the differences were statistically significant (P<0.05). And there was no significant difference in the incidence of adverse reactions in 2 groups (P>0.05). CON-CLUSIONS:Levofloxacin and capreomycin combined with chemotherapy in the treatment of MDR-TB,it can reduce T regulatory cells,increase IL-17 level,do not increase the incidence of adverse reactions.
6.Application of self-made gastroscope outer casing in treatment of esophagogastric variceal bleeding
Zhiyin LI ; Zhonglue WANG ; Gang ZHOU ; Huilin SHEN ; Jumei MA
Chinese Journal of Digestion 2009;29(8):510-513
Objective To estimate the success rate of emergency endoscopic treatment in patients with esophagogastric variceal bleeding using self-made gastroscope outer casing and to investigate bow to prevent complications. Methods A total of 11 patients with esophagogastric variceal bleeding underwent endoscopic treatment with self-made gastroscope outer casing, which consisted of a wire reinforced plastic casing (1.05 cm in inner diameter, 1.35 cm in outer diameter and 50 cm in length), a 3.8 cm×3.8 cm balloon fixed on the casing head and its cavity connected to the gas pipeline placed in casing tail, an anti-reflux pad in the insert entrance of the endoscope tail as well.as a flushing hole. The endoscopy was introduced to the esophagus or stomch together with outer casing to wash and pump the hematocele out from the side of flushing hole. In patients with hemorrhagic lesion in esophageal vein, the balloon was used to press bleeding vein and intravascular sclerotherapy was performed simutanously. In those with hemorrhagic lesion in gastric vein, the balloon was pressed on esophageal vein near to the cardia and intravascular sclerotherapy was performed. Results During the process of examination, the hemorrhagic lesion was fully exposed because of flushing, and bleeding was controlted in all patients after sclerotherapy. No complication such as aspiration or perforation was seen. Conclusions For emergency endoscopic treatment of esophagogastric variceal bleeding, gastroscope outer casing is helpful in removing intragastric hematocele and maintaining a clear field of vision. It can be effectively improving the efficacy of sclerot herapy.
7.Simultaneous determination of tryptophan and kynurenine in plasma by HPLC with UV detection
Li MA ; Mingjun ZHANG ; Qianna ZHEN ; Gang TIAN ; Min DING
Chinese Journal of Laboratory Medicine 2011;34(12):1057-1061
ObjectiveTo establish an accurate method for simultaneous determination of plasma Kyn and Trp by HPLC-UV detection.Methods Kyn and Trp were separated on Agilent Hypersil ODS column using 3-nitrotyrosine as internal standard.The mobile phase consisted of 15 mmol/L sodium acetateacetic acid (pH 5.5):acetonitrile 94∶ 6(v/v) at a rate of 0.8 ml/min.The chromatographic separation was performed at 25 ℃.The eluate was monitored with programmed wavelength setting at 360 nm from 0 to 4 min for Kyn and at 302 nm from 4 to 5 min for Trp.The method was applied to determination of plasma Kyn and Trp in 8 chronic glomerulonephritis,10 idiopathic thrombocytopenic purpura,15 chronic hepatitis B virus patients and 15 healthy controls from September to December in 2010.The differences were compared using ANOVA and SNK methods.Results The retention time of Kyn and Trp were 2.9 min and 4.4 min,respectively.For Kyn,the assay was linear from 0.44 μmol/L to 18.30 μmol/L.For Trp,the linearity was from 3.67 μmol/L to 470.00 μmol/L.The detection limits were 0.014 μmol/L for Kyn and 0.122 μmol/L for Trp,respectively.The within-day CVs were < 3% and the between-day CVs were < 4%.The mean recoveries yield were in the range of 92.29 to 104.40.The plasma concentrations of Kyn were ( 1.59 ± 0.28),(2.73 ± 0.56),(2.69 ± 0.44) and ( 1.54 ± 0.48 ) μmol/L,the plasma concentrations of Trp were (59.8 ± 10.0),(46.1 ± 11.7),(58.5 ±8.0) and (41.4±13.1) μmol/L,the Kyn/Trp were (0.027 4±0.007 5),(0.061 6 ±0.016 5),(0.046 7 ±0.009 1) and (0.038 3 ±0.007 5)in controls,chronic glomerulonephritis patients,idiopathic thrombocytopenic purpura patients and chronic hepatitis B virus patients,respectively.There were significance difference of Kyn,Trp and Kyn/Trp amony the four groups (F=23.734,8.463,20.921,all P<0.01).Conclusion The method is simple,fast,and suitable for applicability to clinical measurement.
8.Laparoscopic distal pancreatectomy
Lei LI ; Dianrong XIU ; Bin JIANG ; Zhaolai MA ; Gang WANG
Chinese Journal of General Surgery 2013;(5):332-336
Objective To study the safety and feasibility of laparoscopic distal pancreatectomy in different modalities.Methods From September 2005 to October 2012,57 patients with masses located at pancreatic body/tail were treated by laparoscopic distal pancreatectomy.According to the operation procedure,patients were divided into four groups:distal pancreatectomy with splenectomy; Spleenpreserving distal pancreatectomy with conservation of the splenic artery and vein; spleen-preserving distal pancreatectomy and without conservation of the splenic artery and vein and laparoscopic to open distal pancreatectomy.Data of each group for operation time,blood loss and complications were collected and statistically analyzed.Results In 57 patients,5 cases were converted to open distal pancreatectomy.52 cases underwent successful laparoscopic operation (including 1 cases of hand-assisted).22 cases of spleen preserving operation (39%),and 30 cases of splenectomy operation (61%).Among the 4 groups of patients operation time,intraoperative blood loss,postoperative length of stay were not significantly different (all P > 0.05).There was no operative mortality.Two cases suffered from B level postoperative pancreatic leakage,there was no C level pancreatic leakage.Conclusions Laparoscopic distal pancreatectomy with or without splenectomy is safe for the treatment of body and tail tumors of the pancreas.
9.Morphology damages of rat articular cartilage induced by different doses of T-2 toxin
Fan-gang, MENG ; Wan-cheng, MA ; Li-hua, WANG
Chinese Journal of Endemiology 2011;30(5):498-501
Objective To study the damage of rat articular cartilage induced by different doses of T-2 toxin, and to explore the relationship between mini-dose T-2 toxin and articular cartilage damage. Methods A total of 120 Wistar rats, weighing 50 - 70 g, were randomly divided into four groups according to their body weights: T-2 toxin group 0(control), 100, 200, 300 μg/kg, 30 rats in each group. Animals in the control group were fed standard rat chow, and animals in the three T-2 toxin groups were fed T-2-toxin-contaminated chow (the dose was 100, 200, 300 μg/kg, respectively). After 6 months, rats were euthanized by ether asphyxiation. The bilateral knee joints were collected and section prepared. The articular cartilage was examined by light and electronic microscope. Results Light microscope showed, the rat articular chondrocytes were clear and arranged orderliness in the control group. The rat articular chondrocytes were disarranged in 100 μg/kg T-2 toxin group.Degeneration and necrosis were found in 200 μg/kg group. Chondrocytes were shrunken with hypereosinophilia cytoplasm and fragmented pyknotic nuclei, extensive areas of chondrocyte loss and chondrocyte clones were visible in 300 μg/kg group. Scanning electronic micrograph(SEM) showed, the rat articular chondrocytes were clear, well formed and arranged tidy in the control group. The surface of articular cartilage was rough in 100 μg/kg group.Collagen fasciculi ruptured and stacked up in 200 μg/kg group. Presented a typical articular dryness phenomenon,the cartilage surface collapsed and many pits appeared in 300 μg/kg group. Transmission electronic microscope (TEM) showed that chondrocytes were abundant with cytoplasm, well-developed rough endoplasmic reticulum in the control group; agglomerate chromatin scattered along the karyotheca, nuclear membrane was thickening, with vacuolar degeneration of the endoplasmic reticulum in the 100 μg/kg group; endoplasmic reticulum expended, with protein retention and organelles breaks in the 200 μg/kg group. A large number of chondrocytes lost organdles, the membrane structures disrupted and the cartilage matrix stromatolyzed in the 300 μg/kg group. Conclusions Within the range of 100 - 300 μg/kg, T-2 toxin induces dose-related articular cartilage injury, the greater the dose, the more serious damage.