1.Clinical study on administration of erythropoietin in patients with acute myocardial infarction
Haipeng GUO ; Yongsheng SONG ; Qizhu TANG
Clinical Medicine of China 2008;24(8):772-774
Objective To study the effect of erythropoietin on cardiac function of the patients with acute mycardial infarction (AMI). Methods 48 patients with AMI successfully treated with thrombolytic therapy were randomized into two group,2000 units of recombinant human erytfu-opoietin(rh-EPO) were administrated once a time in therapeutic group,3 times on alternate days in one week and total for 4 weeks. The peak value of serum creatine kinase(CK) and creatinkinase isozyme MB (CK-MB)were measured, myocardial infarct size (S) was estimated by Hindmen's QRS scoring system, and the diameter of left ventricular end diastolic(LVEDd) and left ventricular injection fraction (LVEF) were determined with echocardiography at the 4th weekend in both groups. Results CK, CKMB and S in therapeutic group were lower than in control group (P < 0.05). LVEDd and LVEF were also improved in the therapeutic group. Conclusion rh-EPO can significantly lessen the size of isehemia and infarct myocardium, mitigate the infarction degree and improve the cardiac function slightly in AMI patients.
2.Effect of Ulinastatin on Anti-inflammation and Pulmonary Function Protection for Infants at Cardiopulmonary Bypass Surgery
Qingling ZENG ; Peijia TANG ; Yuexiu XU ; Yongsheng LIANG
Chinese Circulation Journal 2014;(10):819-822
Objective: To explore the effect of ulinastatin on anti-inflammation and pulmonary function protection with its mechanism for infants at cardiopulmonary bypass surgery. Methods: A total of 38 infants with ventricular septal defect undergoing cardiac operation were randomly divided into 2 groups. Ulinastatin group, the patients received uliastatin 20 000 U/kg,n=20 and Control group, the patients received the same volume of normal saline,n=18. The serum levels of TNF-α, IL-2, IL-l0 were examined at 4 time points: 5 min before skin incision (T1), immediate opening of aorta (T2), 4 hours after operation (T3) and 24 hours after operation (T4). The expressions of CD4+CD45+ T cells and CD4+Foxp3+ T cells were measured at T4. The respiratory index and oxygenation index at 4 time points were compared between 2 groups. Results: Compared with Control group, Ulinastatin group had the lower levels of TNF-α, IL-2 and higher level of IL-l0 at T2, T3, T4; Ulinastatin group also had the higher oxygenation index and lower respiratory index at T2, T3, T4, allP<0.05. Ulinastatin group had less expression of CD4+CD45+ T cells (35.98 ± 3.67)% than Control group (41.94 ± 4.56)% , and more expression of CD4+Foxp3+ T cells (19.65 ± 3.45)% than Control group (6.45 ± 1.47)%,P<0.05-P<0.01. Conclusion: Ulinastatin may improve the differentiation from CD4+CD45+ T cell to Foxp3+CD4+ T cell, down regulating inlfammatory response and protecting pulmonary function for infants at cardiopulmonary bypass surgery.
3.Establishing a luciferase reporter system to evaluate osteogenic differentiation poten-tial of human adipose-derived stem cells
Wenshu GE ; Yiman TANG ; Xiao ZHANG ; Yunsong LIU ; Yongsheng ZHOU
Journal of Peking University(Health Sciences) 2016;48(1):170-174
Objective:Human adipose-derived stem cells (hASCs)are a highly attractive source in bone tissue engineering.To generate a luciferase reporter system that could be used to quantitatively and rapidly examine osteogenic differentiation potential of human adipose-derived stem cells (hASCs ) in vitro,and eventually make it possible to monitor the osteogenic differentiation of transplanted cells in vi-vo.Methods:The genomic DNA harboring promotor regions of osteocalcin and DNA sequences encoding luciferase genes were amplified by PCR and cloned into the pLVX-pTRE-puro vector to generate the OCpro-Luc-Puro construct.Then,the OCpro-Luc-Puro construct together with three assistant vectors:pM-DLg/pRRE,pRSV-REV,and pVSVG,were transiently transfected into HEK293T cells followed by viral supernatants collection,filtration and concentration.Next,the hASCs stably expressing luciferase repor-ter gene driven by osteocalcin promotor were created with the lentivirus carrying OCpro-Luc-Puro cassette under puromycin selection.The OCpro-Luc-hASCs were then cultured in the absence or presence of osteo-genic differentiation medium.On the 7th and 1 4th days,after osteogenic induction,cellular extracts were collected and analyzed by luciferase reporter assay.Meanwhile,alizarin red staining and quantification as well as quantitative reverse transcription PCR (qRT-PCR)analysis of osteogenic associated genes osteo-calcin (OC),runt-related transcription factor 2 (Runx2)and alkaline phosphatase (ALP)were used to assess the osteogenic differentiation ability of OCpro-Luc-hASCs.Results:OCpro-Luc-Puro plasmid and OCpro-Luc-hASCs were successfully generated.On the 7th and 1 4th days after osteogenic induction,the luciferase activity of the cellular extracts from OCpro-Luc-hASCs was dramatically increased.Consistently, the extracellular matrix mineralization,as shown by Alizarin red S (ARS)staining and quantification was also markedly intensified and a marked increase of the mRNA expression levels of OC,Runx2 and ALP, although to variable extent,was observed upon osteogenic differentiation.These results indicated that the observations from traditional experiments examining hASCs osteogenic differentiation were largely in agreement with that of our luciferase reporter assay in OCpro-Luc-hASCs.Conclusion:We established a luciferase reporter system that could be used to rapidly,quantitatively and specifically determine osteo-genic differentiation ability of hASCs.Comparing with the traditional time-consuming methods,the system we generated here was highly effective.This system not only can be used to examine ostogenic differentia-tion of hASCs in a high throughput manner,but also provides a way to monitor ostogenic differentiation of cells in vivo.
4.Effects of adrenomedullin and proadrenomedullin N-terminal 20 peptide, alone or in combination, on the rat hearts in vitro
Jinghui YANG ; Juxiang LI ; Bin GENG ; Yongsheng REN ; Chaosh TANG
Journal of Peking University(Health Sciences) 2003;0(06):-
Objective: To study the effects of adrenomedullin (ADM) and proadrenomedullin N terminal 20 peptide (PAMP) alone or in combinations on the isolated rat hearts as well as the possible signaling pathways involved in their actions. Methods: In isolated rat hearts, the left ventricular pressure (LVP), LVP?dp/dtmax, coronary fluid (CF) and heart rate(HR) of the hearts infused at different concentrations of ADM and/or PAMP were determined by a 4 cannal physiological recorder, then the cAMP contents were assayed in myocardium. Results: After being infused with ADM from 10 -11 to 10 -8 mol?L -1 or PAMP from 10 -11 to 10 -8 mol?L -1 , the LVP and LVP?dp/dtmax of the isolated hearts decreased gradually in a concentration dependent manner, and at the same concentration, the effects of PAMP were more potent than those of the ADM. When ADM and PAMP were co administrated with both concentrations as low as from 10 -11 to 10 -10 mol?L -1 , the cardiac parameters were decreased more than either ADM or PAMP administrated alone. However, the inhibitory effects of ADM and PAMP were attenuated when they were in combination at higher concentrations as from 10 -9 to 10 -8 mol?L -1 . When the rat hearts were infused with ADM, PAMP,and ADM plus PAMP, the CF were always higher than those of the controls and decreased when co administrated with L NAME, an inhibitor of NOS, but the decreaseddegree of LVP and LVP?dp/dtmax were attenuated by L NAME.The cAMP contents in the left cardiac ventricle were increased significantly by ADM infusions but not changed obviously by PAMP, and were of no statistical difference in rat hearts with ADM administrated alone from those combinated with ADM and PAMP. Conclusion: These results showed that ADM and PAMP infused alone or in combinations inhibited the function of rat hearts in vitro, which might be partly involved with the NOS/NO pathway.
5.Relationship between MTHFR gene mutation and keloid
Gang ZHANG ; Yongsheng YE ; Shaojun LUO ; Shaomin TANG ; Jie LIANG
Chinese Journal of Medical Aesthetics and Cosmetology 2008;14(3):163-166
Objective To study the MTHFR gene (677 gene loci and 1298 gene loci) mutations in Chinese patients with keloid. Methods The tissue DNA was extracted from 20 samples of keloids. and peripheral blood samples from the same patients were employed as the control. Polymerase chain reaction(PCR) was used to amplify the Mthfr 677 gene loci and Mthfr 1298 gene loci from the keloid tissue DNA and peripheral blood DNA. and the PCR products were sequenced directly and then compared with the GenBank data. Results Mutations were detected in 17 out of 20 keloids on Mthfr 677 gene loci, the mutation incidence was 85.0 %. Mutations were detected in 13 out of 20 keloids on Mthfr 1298 gene loci, and the mutation incidence was 65.0 %. The mutation involved point mutation, deletion and insetion as well as multisite and multitype. No MTHFR gene mutation was detected in all peripheral blood samples . Conclusion There is a strong correlation between the MTHFR gene (677 gene loci and 1298 gene loci) mutation and keloid.
6.CT Manifestations of Acinar Cell Carcinoma of the Pancreas
Laihua YANG ; Zhongmin TANG ; Yongsheng WANG ; Liangju SHENG ; Dongqing WANG
Chinese Journal of Medical Imaging 2014;(9):686-688,693
Purpose To analysis the CT manifestations of acinar cell carcinomas of the pancreas (ACCs) in order to know more about its CT signs. Materials and Methods The plain and enhanced CT findings of 9 patients with AACs proved pathologically were analyzed retrospectively. Results The main image ifndings of the patients were as follows:①the pancreas grew with exophytic dilatability;②they tended to be large (average diameter was 4.7 cm), with round or oval shape; ③ the lesions showed hypodense on enhanced scan and the solid areas showed slight enhancement in the arterial phase compared with normal pancreas;④most lesions had uniform or partial thin enhanced ring;⑤most lesions demonstrated cystic or necrotic;⑥few had pancreatic/biliary ductal dilatation and peripancreatic involvement;⑦few showed internal calciifcation or intratumoral hemorrhage. Conclusion Plain CT scan and enhanced scan are signiifcant in locating and differentiating acinar cell carcinoma of the pancreas.
7.Trans-activator of transcription protein transduction domain mediated cell membrane penetration of HBcAg
Qingchun PAN ; Guoqing ZANG ; Yongsheng YU ; Zhenghao TANG ; Wei ZHANG ; Jingchao HAN
Chinese Journal of Infectious Diseases 2008;26(6):336-340
0bjective To observe the cell membrane penetration of protein transduction domain (PTD)-HBeAg fusion protein in vitro.Methods The sequence of trans-activator of transcription (Tat)-PTD was synthesized and the whole HBcAg gene was amplified by polymerase chain reaction (PCR).Overlap extension PCR was employed to fuse Tat-PTD and whole HBcAg gene.Then the fusion gene was cloned into prokaryotic expression vector pMAL-c2X.The correct vector was transformed into E.coli Rosetta-gamiTM 2(DE3),and the protein was induced by isopropyl β-D-1-thiogalactopyranoside(IPTG).Western blot was used tO identify the protein. Furthermore,the fusion protein PTD-HBcAg was purified by affinity chromatography.HBcAg protein expressed using the same methods was employed as eontr0l.The purified protein was added tO HuH-7 cell culture,then the transduction of PTD-HBcAg and HBcAg in cells were detected by indirect immunofluorescence assay (IFA).Results The fusion protein was effectively expressed in E. Coli and purified by affinity chromatography.Both purified PTD-HBcAg and HBcAg could be recognized by HBeAg monoclonal antibody in Western blot analysis.IFA visualization showed that PTD-HBeAg could be introduced into HUH-7 ceils while HBcAg only could not be detected in cells.Conclusions PTD-HBcAg fusion protein can be expressed effectively and purified in prokaryotic expression system.PTD could mediate HBcAg penetrating eell membrane into the cells.
8.Clinical study on 3D-CRT combined with nimotuzumab and chemotherapy in the treatment of stages Ⅲ-ⅣA nasopharyngeal carcinoma
Wubing TANG ; Wen YANG ; Yang CAO ; Churong WU ; Xingxi PAN ; Zhenheng LIU ; Yongsheng ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2012;19(9):1293-1295
Objective To explore the effect and safety of three-dimensional conformal radiotherapy (3D-CRT) m combination with nimotuzumab and chemotherapy in treatment patients with locoregionally advanced nasopharyngeal carcinoma.Methods 60 patients with stages Ⅲ-ⅣA nasopharyngeal carcinoma were enrolled.All patients were treated with 3D-CRT and concurrent and sequential chemotherapy by paclitaxel and eisplatin and given nimotuzumab 100mg i.v.weekly for 6 ~ 7 weeks before radiotherapy.Results After two months,the complete response rates(CR) of nasopharyngeal carcinoma and cervix lymph nodes were 98.3%,96.7% respectively,l-year locoregional control and distant metastasis-free survival rates were 100%,96.7% in 38 patients.2-year,3-year locoregional control and distant metastasis-free survival rates were 100% in 16 patients and 8 patients.The major side effects included oral mucositis,actinodermatitis,neutropenia,nausea,vomiting,and fatigue.Grade 3 acute oral mucositis often occured.No skin rash or allergic toxicities appeared.All the effects were tolerable.Conclusion 3D-CRT combined with nimotuzumab and peclitaxel and cisplatin chemotherapy may improve the complete response rate and locoregional control and distant metastasis-free survival rate on locoregionally advanced nasopharyngeal carcinoma,with mild to moderate side effects.
9.Associating liver partition and portal vien ligation for staged hepatectomy
Jian ZHOU ; Zheng WANG ; Jian SUN ; Yongsheng XIAO ; Jia FAN ; Zhaoyou TANG
Chinese Journal of Digestive Surgery 2013;(7):485-489
Complete resection of liver cancer is the main approach for achieving radical resection,and sufficient remnant liver is essential for avoiding hepatic failure after operation.With the aim of increasing remnant liver volume,a new two-stage technique,associating liver partition and portal vein ligation for staged hepatectomy (ALPPS),recently has been developed.In this article,the initial experience with 1 case of hepatocellular carcinoma who underwent ALPPS at the Zhongshan Hospital in April 2013 was reported.In the first stage,the right portal vein branch was ligated and subsequently the liver parenchyma was dissected along the falciform ligament to isolate the segment Ⅳ and the left lateral lobe.On postoperative day 7,the remnant liver volume was increased from 291 ml to 579 ml,and on postoperative day 8,the second stage operation was performed.During the second stage,the extended right lobe was removed.ALPPS induces a great and fast hypertrophy of the remnant liver,and R0 resection could be performed on patients which was considered unresectable because of small remnant liver volume,without severe postoperative liver failure and has a low mortality.
10."The Treatment Experience of""Front-type""Hypospadias (40 Cases Reports)"
Yongsheng CHEN ; Siman LI ; Mingdong PENG ; Cheng WANG ; Haiyu JU ; Wenguang TANG
Journal of Kunming Medical University 2014;(2):12-14
Objective To explore the operation points of the steps offront-typehypospadias operation in oral mucosa urethroplasty and scrubbed shaped urethra with meat membrane covering.Methods After correction of chordee of penis of 40 patients with Front -type hypospadias, oral mucosa was transplanted and fixed on albuginea surface at the one-third of ventral penile for all the patients to increase the width of the urethra and form the urethra with the selected appropriate size ureter. The skin of dorsal penile was transferred to ventral penile. After clearing the pedicled skin flap, the subcutaneous layer of meat was kept down, and stamped wholly on forming place of urethral reel (including both sides inferior of cut-off cavernous body of glans penis),forming the glans again.Results There was no ankylo-urethria among the 40 front-type hypospadias operation, ureteroscopy examination after two months of the operation showed that all the transplanted oral mucosa survived, and the stamped subcutaneous layer of meat located at both sides inferior of cut-off cavernous body of glans penis adhered with satisfaction,no glans incision dehiscence,there occurred 2 cases of urinary fistula which had been cured by neoplasty,there was 1 case of transferred flap necrosis which had been cured after dressing change.40 patients were satisfied with penis appearance after operation.Conclusion Following up the operation points of “front-type”hyospadias operation,the success rate of operation can be improverd obviously,the plastic effect is good,and the complications after operation can be reduced.stamped wholly on formed urethra.There is a small probability of incidence of urethral stenosis and urinary fistula after operation.