1.Construction of BMP_(4/7) fusion gene adeno-associated virus and biological effects of transfection on bone marrow stromal cells in rabbits
Shaohui YUAN ; Jian SHANG ; Guojun SHAO
Orthopedic Journal of China 2006;0(20):-
[Objective]To determine the biological effects of transfection of adeno-associated rirus(AAV) BMP4/7 on rabbit bone marrow stromal cells (BMSCs).[Method]The mature peptide of BMP4 and BMP7 were gained by One-Step RT-PCR from the human placenta and the BMP4/7 fusion gene was gained through gene recombinant techniques and then transferred to pGEM plasmid. The BMP4/7 fusion genen was cut down from the pGEM-BMP4/7 and the recombination was successfully completed in colibacillus, and recombinant adeno-assosiated was produced in 293 cells. Rabbit BMSCs were transfected with the recombinant adeno-assosiated virus vectors carrying BMP4/7 fusion gene with differen multiplicitv of infection(MOI) values. Cell growth curves were drawn to evaluate the biologic effect of AAV-BMP4/7 on cyto-activity. The transfection efficiency was measured using MTT method. The ossification of cells was evaluated by investigating the shape change of the cell ability of ALP and OC at 7,14 days after transfection. Cells were then transfected with AAV-BMP4/7 and AAV-EGFP,respectively.[Result]1.We successfully constructed the recombinant adeno-assosiated virus with BMP4/7 fusion gene.2.The ttransfection efficiency of AAV-BMP4/7 was roughly 72% without siginifficant biologic effect on cyto-activity.The ossification of cells was significant after transfection with AAV-BMP4/7. The 1?105 vg/cell MOI value of transfection efficiency was stroger than 5~104 vg/cell MOI value (59.3,8%). 3. The concentrations of ALP and OC in AAV-BMP4/7 transfection groups were significantbly higher than in AAV-EGFP groups (tALP=896.88 P
2.The Comparative Study of Oral Resonance Acoustic Characteristics of School -Aged Children with Down Syndrome and Normal Children
Qin WAN ; Guojun SHAO ; Zhaoming HUANG
Journal of Audiology and Speech Pathology 2013;(5):469-473
Objective To compare the resonance acoustic characteristics between 7~16 years old Down syn-drome children and normal children .Methods A total of 29 cases of 7~16 years old Down syndrome children and 90 cases of 7~16 years old normal children pronuanced /a/,/u/,/i/,each sound for 3 times ,every time 1 to 2 sec-onds .The oral resonance acoustic parameters ,including F1 (a) ,F2 (a) ,F1 (i) ,F2 (i) ,F1 (u) ,F2 (u) ,the distance of tongue and the distance of jaw ,were recorded by real analysis .Analysis of variance was used for the voice acous-tic .Results With age ,there was a significant decrease tendency of the value of F2 (a) and a very significant growth tendency of the values of F2 (i) and the distance of tongue in normal children .The value of F2 (i) inrcreased signifi-cantly in Down syndrome children .The values of every oral resonance acoustic parameters were not different signifi-cantly on sex in normal children .The values of F1 (i) and F1 (u) in Down syndrome boys were significantly less than those of in the Down syndrome girls ,but the the distance of jaw in Down syndrome boys were significantly greater than those of in the Down syndrome girls .The values of F2 (u) and F1 (i) in Down syndrome children were signifi-cantly greater than those of in normal children ,but the distance of tongue was significantly less than those of in nor-mal children .Conclusion With age ,in normal children ,the amplitude of the tongue forward movement and the range of tongue back and forward movement were increased significantly ,the oral focus moved forward gradually .In Down syndrome children ,the amplitude of the tongue back movement ,and the range of tongue back and forward movement ,were poor than those of in normal children .The Down syndrome children were easier to suffer from oral resonance disorder than normal children .
3.Expression, purification and characterization of a thermostable lactate dehydrogenase from Thermotoga maritima.
Guojun QIAN ; Caiping CHEN ; Ruying ZHAI ; Weilan SHAO ; Yanzhen MEI
Chinese Journal of Biotechnology 2014;30(4):545-553
The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular
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Enzyme Stability
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Escherichia coli
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metabolism
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L-Lactate Dehydrogenase
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biosynthesis
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Molecular Weight
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Recombinant Proteins
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biosynthesis
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Temperature
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Thermotoga maritima
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enzymology
4.Epidemiological Situation of Sexually Transmitted Diseases in China:from 1991 to 2001
Xi-Angdong GONG ; Shunzhang YE ; Junyan ZHANG ; Guocheng ZHANG ; Changgeng SHAO ; Guojun LIANG ; Wen-Hua JIANG ; Qiang XIA ; Quanpei WANG
Chinese Journal of Dermatology 1995;0(03):-
ObjectiveTounderstandthetrendsandepidemiologicalcharacteristicsofsexuallytransmitteddiseases(STDs)inChinaandprovidescientificbasisformakingcontrolstrategies.MethodsDuringtheperiodof1991~2001,thecase-reportingdataof8kindsofnotifiableandmonitoringSTDs,collectedfrom31provinces,autonomousregionsandmunicipalities,wereanalyzedwithepidemiologicalmethods.Results①Epidemictrends:Duringthisperiod,theincidenceof8kindsofSTDssteadilyincreasedfrom175528(15.48per100000population)in1991to859040(68.91per100000population)in2000.Theaverageannualgrowthofincidencewas19.30%,witharangeof2.59%~36.88%.However,thereportedcasesin2001were795612withadecreaseof7.38%comparedwiththosein2000,anditwasthefirstdecreasesinceSTDcaseswerereportedfrom1987.②Geographicaldistribution:Thehigh-incidenceareasweretheZhujiangRiverDelta,YangtzeRiverDelta,MinjiangRiverValley,NortheasternChina,andBeijing,Tianjin,andChongqingManicipalities,withtheincidencerateofover70~100casesper100000populationafter1997,andtherewereveryhighratesofincidenceover1000casesper100000populationinsomeareas.Thelow-incidenceareaswerenorthChina,partsofCentralChina,NorthwesternChinaandSouthwesternChina,withtheincidenceratesoflowerthan30~50per100000population.③Populationdistri-bution:Themaletofemaleratiodecreasedfrom1.60∶1~1.69∶1intheearly1990sto1.35∶1~1.40∶1inthelate1990s.STDincidencerateswerehighestinthe20~39agegroup,andthereportedSTDcasesofthisagegroupaccountedforover80%oftotalcases.ConclusionSexuallytransmitteddiseasesinChinahavebecomeaseriouspublichealthproblemandtheeffectiveinterventionprogrammesagainstSTDsmustbeimplementedacrossthecountry.
5.Simultaneous Determination of 8 Components in Total Flavanones of Sedum sarmentosum Bunge by UPLC-MS/MS
Xue WANG ; Zhitao JIANG ; Guojun YAN ; Zhuying SHAO ; Xingyue FENG ; Jinhuo PAN
China Pharmacy 2018;29(9):1222-1226
OBJECTIVE:To establish a method for simultaneous determination of hyperoside,quercitrin,luteoloside, kaempferol,quercetin,rutin,luteolin and isorhamnetin in total flavanones of Sedum sarmentosum Bunge. METHODS:UPLC-MS/MS method was adopted. The determination was performed on ZOBAX SB C18column with mobile phase consisted of methanol-5 mmol/L ammonium formate aqueous solution(45:55,V/V)at the flow rate of 0.4 mL/min. The column temperature was 30 ℃, and sample size was 2 μL. The electrospray ionization source(ESI)was used;ion source temperature was 400 ℃;desolvation temperature was 300 ℃;desolvation gas flow was 600 L/h;capillary voltage was 3 000 V;nebuliser pressure was 45 psi;the work mode was multiple reaction monitoring mode;detection mode was negative ion mode. The established method was used to determine the contents of 8 components in 3 batches of total flavanones of Sedum sarmentosum Bunge. RESULTS:The linear ranges of hyperoside,quercitrin,luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin were 10.0-640.0,0.5-32.0, 4.5-288.0,8.0-512.0,50.0-3 200.0,2.0-128.0,12.5-800.0 and 25.2-1 612.8 ng/mL(r≥0.991 4),respectively. The limits of detection were 5.0,0.25,2.25,4.0,25.0,1.0,6.25 and 12.6 ng/mL,respectively.The limits of quantitation were 10.0,0.5,4.5, 8.0,50.0,2.0,12.5 and 25.2 ng/mL,separately. RSDs of precision,stability(24 h)and reproducibility tests were no more than 4.3%(n=6). The recoveries were 95.9%-100.6%,and RSDs were 1.5%-3.8%(n=6). The contents of hyperoside,quercitrin, luteoloside,kaempferol,quercetin,rutin,luteolin and isorhamnetin in 3 batches of total flavanones of Sedum sarmentosum Bunge were 507.88-560.37,42.95-50.36,63.52-71.80,1 695.10-1 753.27,10 569.28-10 612.99,25.76-30.13,2 795.22-2 877.43 and 4 869.55-4 971.30 μg/g,respectively. CONCLUSIONS:The established method can be used for simultaneous determination of 8 components in total flavanones of Sedum sarmentosum Bunge.
6.Cause analysis and clinical management of postoperative complications of proximal femoral nail anti-rotation in treating intertrochanteric fractures
Tienan SONG ; Xuming WANG ; Xiaojun YU ; Guojun LI ; Yinghao LI ; Ming SHAO
Chinese Journal of Geriatrics 2018;37(12):1324-1327
Objective To investigate the causes and managements of postoperative complications of proximal femoral nail anti-rotation(PFNA)in treating intertrochanteric fractures. Methods A total of 155 patients with intertrochanteric fractures were treated with PFNA from January 2011 to December 2017.Causes and managements of postoperative complications were analyzed and discussed. Results At an average follow-up of 48 months(range ,12~72 months) ,the rate of good postoperative recovery in hip function was 96.0% . Among 155 patients ,the internal fixation-associated postoperative complications occurred in 15(9.7% )patients.Details were as follows :7 cases had spiral blade breakage into the femoral head or inwardly ,in whom 5 cases received artificial joint replacement whereafter.5 cases had coxa vara ,mainly in patients with obvious posterior fracture of the femoral intertrochanter with obviously bone fragments.3 cases had delayed fracture healing due to the fracture damaging calcar femorale ,or extensive soft tissue dissection during the operation on small trochanter. The Harris function score was 82 points. Conclusions The location of the PFNA spiral blade ,as an important factor ,is correlated with postoperative complications ,while severe osteoporosis ,unstable fractures ,and poor fracture reduction increase the incidence of surgical complications.
7. Epidemiological study of occupational diseases in Gansu province, China in 2010-2017
Wenli ZHAO ; Pingtai LIAO ; Zhenxia KOU ; Yuhong HE ; Xiaogang HE ; Guojun SHAO ; Jia XU ; Hui LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(10):789-792
Objective:
To analyze the epidemiological characteristics of occupational diseases in Gansu Province, China in 2010-2017, and to provide a scientific basis for the prevention and control of occupational diseases.
Methods:
The cluster sampling method was adopted to make statistical analysis of 1339 cases of occupational disease reported by "occupational disease and occupational health information monitoring system" in Gansu province from 2010 to 2017, to investigate the diseases, regions and industries of occupational diseases in June 2018.
Results:
A total of 1339 cases of occupational diseases (39 types, 8 classes) were diagnosed and reposed in 2010-2017. The three most frequent diseases were pneumoconiosis (87.53%, 1172/1339), occupational poisonings (5.83%, 78/1339), and occupational ear, nose, and throat (ENT) diseases (3.14%, 42/1339). The cases of silicosis accounted for 54.61% (640/1172) of all cases of pneumoconiosis, the second was coalworker pneumoconiosis, which accounted for 38.57% (452/1172). In the cases of occupational poisonings, 32.05% (25/78) suffered from carbon monoxide poisoning. Patients with occupational diseases were reported in 14 districts of Gansu, mostly in Lanzhou (27.52%, 347/1261), Jinchang (16.57%, 209/1261), and Baiyin (14.20%, 179/1261). The reported cases are mainly concentrated in mining (71.56%, 468/654) and manufacturing (21.87%, 143/654), the types of state-owned economy (55.63%, 692/1244) and private economy (33.68%, 419/1244), large (43.41%, 540/1244) and small enterprises (35.21%, 438/1244) in 2010-2017 in Gansu.
Conclusion
The pneumoconiosis caused by silicious and coal dust and the occupational poisonings caused by carbon monoxide seem to be the main occupational hazards in Gansu province. Occupational diseases occur in all districts of Guangzhou and in various industries. The state-owned economy and private sector, large and small enterprises should be the focuses of occupational health supervision.
8.Preliminary evaluation of sunitinib as first line therapy in treating patients with metastatic renal cell carcinoma
Jianlin YUAN ; Fuli WANG ; Weijun QIN ; Jun QIN ; Xiaojian YANG ; Chunjuan TIAN ; Rongliang QIN ; Chen SHAO ; Lijun YANG ; Fei LIU ; Geng ZHANG ; Ping MENG ; Longlong ZHANG ; Yu ZHENG ; Guojun WU
Chinese Journal of Urology 2015;(10):742-745
[Abstact] Objective To investigate the efficacy and safety of sunitinib as first line therapy in treating those patients with metastatic renal cell carcinoma ( mRCC ) .Methods A total of 66 patients , including 42 male and 24 female cases ,with metastatic renal cell carcinoma were enrolled from January 2009 to June 2014.The median age was 52 years (range 26-75 years).According to American Joint Committee On Cancer (AJCC) staging,there were 35 cases of T3 stage,31 cases of T4 stage.All patients had distant metastasis ,including single organ metastasis in 52 patients and multiple organ metastasis in 14 cases.Sixty-one patients received prior radical nephrectomy ,5 patients received biopsy .Sixty-two patients were diagnosed as renal clear cell carcinoma and 4 patients were diagnosed as renal papillary cell carcinoma .Sunitinib was administered in standard 4/2 regimens.Briefly, patient takes 50 mg once a day orally for 4 weeks.Then the sunitinib will be stopped for 2 weeks.Six weeks was defined as 1 cycle.It should be continued until disease progression or occurrence of intolerable adverse reactions .The efficacy of sunitinib should be evaluated within 2 cycles.Results The duration of following-up ranged from 5 to 66 months.The efficacy could be evaluated in 63 patients.Two patients ( 3.2%) achieved complete remission .Twelve patients ( 19.0%) achieved partial remission.Forty-five patients (71.4%) demonstrated stable disease and 4 patients (6.3%)
developed progressive disease .The disease control rate was 93.7%(59/63) and the objective response rate was 22.2%(14/63).2 (3.2%) patients died due to the progression of disease .The most commonⅠ-Ⅱadverse events included fatigue in 36 cases ( 57.1%) , thrombocytopenia in 36 cases ( 57.1%) , hand-foot syndrome in 32 cases (50.8%),hypertension in 27 cases (42.9%),neutropenia in 15 cases (23.8%), hypothyroidism in 12 cases (19.0%), diarrhea in 6 cases (9.5%) and alopecia in 4 cases (6.3%).Ⅲ-Ⅳ adverse events were hand-foot syndrome in 4 cases ( 6.3%) , hypertension in 2 cases ( 3.2%) , neutropenia in 5 cases (7.9%) and thrombocytopenia in 5 cases (7.9%).Most mild adverse reactions after symptomatic treatment could be alleviated ,did not affect the medication .When the adverse events returned to the Ⅰ-Ⅱdegree, the 37.5 mg sunitinib was resumed once daily by orally.NoⅢ-Ⅳadverse events were reported again.Conclusions Sunitinib was efficacious in the treatment of advanced renal cell carcinoma.Most mild adverse events were tolerable ,and severe adverse events need medical treatment .
9.Study on the accuracy of oxygen concentration of modified oxygen treatment with Venturi and humidity system.
Qiang WEI ; Bingyu QIN ; Guojun HE ; Yuanyuan WU ; Yuan SHI ; Weitao SUN ; Mengjuan JING ; Shichao ZHU ; Huanzhang SHAO
Chinese Critical Care Medicine 2018;30(7):677-680
OBJECTIVE:
To verify the accuracy of oxygen concentration (FiO2) of modified oxygen treatment with Venturi and humidity system.
METHODS:
Patients just after ventilator weaning and before the removal of tracheal intubation/tracheotomy tube, who admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from May 1st to December 15th in 2017, were enrolled. All patients were given a modified oxygen treatment with Venturi and humidity system, and the oxygen flow rate (Flow) of the Venturi device and the oretical value of FiO2 were adjusted according to the patient's condition. Patients were divided into five groups based on doctor's orders: Flow 3 L/min FiO2 0.24, Flow 3 L/min FiO2 0.26, Flow 6 L/min FiO2 0.28, Flow 6 L/min FiO2 0.30, Flow 9 L/min FiO2 0.35. The value of FiO2 at the inhalation end of patients of each group was measured by TSI airflow analyzer, and the consistency between the measured value of FiO2 at the inhalation end and the FiO2 marked value of Venturi was compared and analyzed.
RESULTS:
When the FiO2 theoretical value of Venturi were adjusted to 0.24, 0.26, 0.28, 0.30, and 0.35, the measured values of FiO2 at the inhalation end of patients were 0.38±0.05, 0.38±0.05, 0.40±0.04, 0.41±0.04, and 0.77±0.11, respectively, which were all significantly higher than the theoretical value of FiO2 (all P < 0.01). The difference between the measured value of FiO2 at the inhalation side and the FiO2 value of the Venturi annotated and the difference rate were both "V"-shaped, both of which decreased with the increase in theoretical value of FiO2 to a Flow of 9 L/min and a theoretical value of FiO2 0.35, the accuracy was the worst, with the FiO2 difference of 0.42±0.11, and the FiO2 difference rate of (121.6±36.5)%.
CONCLUSIONS
There is a difference between the measured value and the theoretical value of FiO2 at the inhalation end of the modified Venturi oxygen therapy humidification system, which needs to be paid attention to during clinical oxygen therapy.
Humans
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Humidity
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Oxygen/analysis*
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Oxygen Inhalation Therapy
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Respiration, Artificial
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Ventilator Weaning
10. Study on the accuracy of oxygen concentration of modified oxygen treatment with Venturi and humidity system
Qiang WEI ; Bingyu QIN ; Guojun HE ; Yuanyuan WU ; Yuan SHI ; Weitao SUN ; Mengjuan JING ; Shichao ZHU ; Huanzhang SHAO
Chinese Critical Care Medicine 2018;30(7):677-680
Objective:
To verify the accuracy of oxygen concentration (FiO2) of modified oxygen treatment with Venturi and humidity system.
Methods:
Patients just after ventilator weaning and before the removal of tracheal intubation/tracheotomy tube, who admitted to the intensive care unit (ICU) of Henan Provincial People's Hospital from May 1st to December 15th in 2017, were enrolled. All patients were given a modified oxygen treatment with Venturi and humidity system, and the oxygen flow rate (Flow) of the Venturi device and the oretical value of FiO2 were adjusted according to the patient's condition. Patients were divided into five groups based on doctor's orders: Flow 3 L/min FiO2 0.24, Flow 3 L/min FiO2 0.26, Flow 6 L/min FiO2 0.28, Flow 6 L/min FiO2 0.30, Flow 9 L/min FiO2 0.35. The value of FiO2 at the inhalation end of patients of each group was measured by TSI airflow analyzer, and the consistency between the measured value of FiO2 at the inhalation end and the FiO2 marked value of Venturi was compared and analyzed.
Results:
When the FiO2 theoretical value of Venturi were adjusted to 0.24, 0.26, 0.28, 0.30, and 0.35, the measured values of FiO2 at the inhalation end of patients were 0.38±0.05, 0.38±0.05, 0.40±0.04, 0.41±0.04, and 0.77±0.11, respectively, which were all significantly higher than the theoretical value of FiO2 (all