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.

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.

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.

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.

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.

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.

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.

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.

Objective To evaluate MRI in judging the therapeutic effect of argon-helium cryoablation therapy for prostate cancer. Methods The clinical data and imaging materials of 16 patients with prostate cancer , who had received ultrasound-guided argon-helium knife cryogenic treatment at authors ’ hospital during the period from March 2012 to Oct. 2014, were retrospectively and comprehensively analyzed. The preoperative and postoperative laboratory test results were compared with MRI findings, and the ablation effect was assessed, focusing on the surgical residue, metastasis, etc. Results One months after the surgery, MRI demonstrated that satisfactory ablation extent was obtained in all patients, the ablated tumor tissue was characterized by long-T2 signal and no obvious recurrence could be found on DWI and DCE-MRI. A slight decrease of PSA level was observed. Six months after the surgery, MRI revealed that the prostate size was significantly reduced, PSA level was markedly decreased and no obvious evidence of recurrence was observed. No severe postoperative complications, such as urethral necrosis, urethral stricture or urethro-rectal fistula, occurred. During the 6-month following-up time, one patient died from other causes. Conclusion MRI has excellent clinical application value in estimating the ablation extent and in judging postoperative recurrence or metastasis of prostate cancer after argon-helium cryoablation treatment.

Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) ％] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )％] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)％] and medium group [(0.09±0.02)％,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.