Retroviral expression system which consists of retroviral vector,envelop protein vector and packaging cell line is an efficient expression system for recombinant protein.It has great potential in gene therapy and biopharmacy.Transcriptional active genome regions are the preferred targets for retrovirus integration.Furthermore,VSV-G protein enables this system a broader host range and makes virus integration more efficient.After infection of high-titer virus,high production clones can be selected through simple screening.So far,the research on retrovirus expression system has developed into application in bio-pharmacy industry.Here the composition of this system and the mechanism of virus transduction and summarize the application and prospect of retroviral expression system are introduced.

This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

Objective To explore the incidence and risk factors of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods The clinical data of 72 cases allo-HSCT from Oct 2004 to Dec 2008 were analyzed. Thirteen factors possibly correlated with the development of aGVHD were analyzed. Results aGVHD was developed in 32 cases (44.4 %), in which grades Ⅰ aGVHD was 11.1%, gradesⅡaGVHD was 18.1%, and grades Ⅲ-Ⅳ aGVHD was 15.3 %. The univariate analysis showed that diagnosis, the status of disease, use ATG, conditioning regimen, donor type,ABO blood group disparity between donor and recipient, CD34+ cell number, early engraftment and neutropenic infection, HLA locus were associated with the occurence of aGVHD (P ＜0.1). On the COX regression mode, an increased risk of aGVHD was associated with HLA mismatch (HR =2.58, P ＜0.005), GVHD prophylaxis without ATG (HR =2.94, P ＜ 0.001), and unrelated donor (HR =1.97, P ＜0.01). Conclusion aGVHD is a common complication after allo-HSCT, and HLA mismatch and unrelated donor are independent risk factors for aGVHD.

Objective To evaluate risk factors of the prognosis in acute arterial embolism of the upper extremities. Methods The clinical data of 62 consecutive patients admired in our hospital with the diagnosis of acute arterial embolism in the upper extremities, from July 1988 to January 2008, were retrospectively reviewed. The risk factors including age, gender, cardiac function, location of embolism, embolectomy and duration of iaehemia were analyzed by cumulative Loots regression. Results There were 62 patients, 33 men and 29 worsen, with a mean age of 63. 5 years (35～86 years). Among them, 37 patients received Fogarty embolectomy and 25 patients received medical treatment including thrombolysis, anticoagulation and antiplatelet therapy because of poor risk for surgery. The iachemic status meliorated in 55 patients (88.7%) with 2 patients receiving amputation and 2 patients dying during the peri-operative period. The result of cursulative Logits regression shewed that the duration of ischemia, cardiac function and embolectomy played the significant role on the prognosis (P < 0.01 ), but age, sex and the location of embolism did not show the significant role. In those only receiving the medical treatment, 23 patients restored blood flow to different degree. Conclusions Embolectomy with Fogarty catheter within 8 hours of onset was the most effective treatment for acute arterial embolism in the upper extremities. For peor-risk patients, early medical treatment including thrombolysis, antieoagulation and antiplatelet treatment, can also restore the blood flow in the isehemie limbs.

Objective To report our surgical experience in treating 19 cases of solitary iliac aneurysms (SIA). Methods The clinical data of 19 consecutive patients with SIA between January 1985 and January 2008 were retrospectively reviewed. There were 18 men and 1 woman, aging from 39 to 77 years ( mean 62 ± 7 years). Results There were 30 SIAs in the 19 patients, including 25 ( 83.3% ) common iliac aneurysms, 4 (13.3%) internal ihac aneurysms and 1 (3. 3% ) external iliac aneurysm. Eleven patients ( 57.9% ) had multiple ancurysms, with 9 patients ( 47.4% ) having bilateral SIA. Two patients had coexistent peripheral vascular occlusive disease. There were 2 patients suffering form ruptured SIA, one was saved by emergency operation and one died before an surgery could be attempted. Seventeen patients underwent successful open aneurysmectomy and artificial graft implantation leaving no ischemic complications of the pelvic organs. One patient with right common iliac aneurysm underwent endovascular repair without endoleak. There was no operative death during porioperative period. The surviving patients remained stable and had good patency of grafts during the follow-up period. Conclusions Early management of SIA is important, CT angiogarphy (CTA) is necessary not only to evaluate the SIAs, but also to detect multiple aneurysms or arterial occlusive disease. Close and long-term follow-up is mandatory for the early detection of the formation of new anearysms.

Objective To investigate the feasibility,security,indication and prophylactic measures of postpartum hemorrhage during the fetalintrapartum operation by analyze the maternal prognosis after intrapartum fetal operation(IFO) performed.Methods We performed a retrospective evaluation of 10 cases that has been performed intrapartum fetal operation(IFO group,n=10) successfully between August 2008 and October 2009 at Department of Obstetrics and Gynecology,Shengjing Hospital,China Medical University.Healthy pregnant women(n=10) were chosen as control group,who received Caesarean section for refusing to vaginal delivery.Between the two groups,we compared the indexes including blood lose during the operation and 24 hours post-operation,the hemoglobin level pre-operation and 24 hours post-operation,the involution of uterus,the puerperal infection,and the follow-up at 42 days post-operation.Results Good involution,no primary and secondary postpartum hemorrhage and no puerperal infection were found in both of the 2 groups.Intra-operation,24 hours post-operation and the total volumes of blood lose,the hemoglobin level before operation,24 hours post-operation and 42 days post-operation had no statistic significant difference between the IFO group and control group(P>0.05).No significant difference was found in the operation time between IFO group and the control group(66.40±53.40 minutes and 34.50±4.97 minutes,respectively.Intra-operation,24 hours post-operation and the total volume of blood lose had no relation to the maternal age and gestational weeks(P>0.05).The blood lose of intra-operation in IFO group had positive linear correlation to the operation time.(Pearson relation coefficient R=0.458,.P=0.021).The linear recurrence equation was:the volumes of blood lose intra-operation(ml)=172.68+1.342x(operation time).Conclusion IFO was safe to the mother.It did not affect the involution of uterus and could not increase the blood lose of postpartum and the puerperal infection morbidity.Through the effective measures of preventing postpartum hemorrhage,the postpartum blood lose of the patients that performed intrapartum fetal operation could be no more than that of the patients that had normal Caesarean section.Improving the technique of the intrapartum fetus operation and shortening the operation time are important to prevent postpartum hemorrhage.

Deficiency Diseases ; prevention & control ; Goiter ; chemically induced ; Humans ; Iodine ; administration & dosage ; adverse effects ; deficiency

Objective The aim of this study was to investigate the effect of miR-129 on the proliferative activity and apop-tosis of osteosarcoma MG-63 cells. Methods Thirty cases of osteosarcoma and its adjacent paracancerous tissues were collected. RT-PCR was used to detect the expressions of miR-129 mRNA and SEPHS1 mRNA. Western blot was used to detect the protein level of SEPHS1. After MiR-129 was over-expressed and knocked down,the cell proliferation was detected in MG-63 cells by CCK-8,and apoptosis was detected in MG-63 cells by Westerm blot and hoechest staining. Results In this study,the expression of miR-129 in osteosarcoma tissues was significantly higher than that in para-cancerous tissues(P<0. 05). Through the established model in vitro of miR-129 overexpression or knockdown,it was determined that the miR-129 mimetic and inhibitor concentrations were optimal for overexpression and knockdown at 50 nM and 200 nM, respectively. The possibility of binding between miR -129 and SEPHS1 was predicted by software,and the luciferase reporter assay further confirmed the binding relationship between miR-129 and SEPHS1. Knockdown of miR-129 significantly inhibited the proliferation of MG-63 cells and accelerated the apoptosis of MG-63 cells. Conclusion miR -129 plays a key role in regulating the proliferation and apoptosis of osteosarcoma cells by binding to SEPHS1.

Objective To explore the value ofneuroendoscope in microvascular decompression for treatment of primary hemifacial spasm. Methods One hundred and eighteen patients with hemifacial spasm, admitted to our hospital from December 2006 to December 2009, underwent endoscope-assisted microvascular decompression via suboccipital retrosigmoid approach; their clinical data and treatment efficacy were retrospectively analyzed. Results The offending vessels were found in 106 patients (89.83%) under microscope, of which 7 (5.93%) were again found other offending vessels under neuroendoscope. Eight patients (6.78%) were noted as no significant vascular compression under microscope, but the offending vessels of these patients existed under endoscope. Seventeen patients performed decompression with Teflon cotton under microscope needed modification when they were checked under endoscope. The symptoms of 102 patients (86.44%) disappeared immediately after the operation; 11 (9.32%) disappeared gradually; 3 (2.54%) had partial remission and 2 (1.70%) had no significantly remission. No serious complications, including cerebrospinal leakage, paralysis and death, were noted. Conclusion Endoscope-assisted microvascular decompression, by improving the observation of the anatomic neurovascular relation, decreasing the damage to the brain stem, nerves and important vessels, can improve the surgical treatment, and reduce the recurrence rate and postoperative complications.

OBJECTIVETo study the state of oxidative stress in patients with acute coxsackie virus myocarditis (ACM), and to investigate the pathological chain reactions of a series of free radicals and oxidative and lipoperoxidative damages in their bodies.

METHODSEighty ACM patients and 80 healthy adult volunteers (HAV) were enrolled in a case-control study, in which concentrations of nitric oxide (NO) in plasma, lipoperoxides (LPO) in plasma and LPO in erythrocytes (RBC), vitamin C (VC), vitamin E (VE) and beta-carotene (beta-CAR) in plasma as well as activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in RBC were determined by using spectrophotometric assays.

RESULTSCompared with the average values (AV) of the above biochemical parameters (BP) in the HAV group, the AV of NO in plasma, and LPO in plasma and RBC in the ACM group were significantly increased (P = 0.0001), while the AV of VC, VE, beta-CAR, SOD, CAT and GSH-Px in the ACM group were significantly decreased (P = 0.0001). The values of the above BP were used to estimate the relative risk ratio (RR) between the ACM group and the HAV group; the RR and its 95% confidence interval were 12.467 (5.745-27.051), 4.333 (2.126-8.834), 6.517 (3.225-13.618), 3.310 (1.598-6.858), 31.000 (12.611-76.201), 4.663 (2.228-9.759), 11.769 (5.440-25.462), 3.043 (1.486-6.229) and 6.594 (3.045-14.281) respectively, and their P levels ranged from 0.002 to 0.0001. The results were as follows: D = 22.143 - 0.017SOD + 0.008NO + 0.244LPO in RBC, Eigenvalue = 13.659, Canonical correlation = 0.965, Wilks' lambda = 0.068, chi 2 = 420.212, P = 0.0001. The correct rate of discrimination to the ACM group and to the HAV group was 87.5% and 95.0%, respectively, and 91.3% of originally grouped cases was correctly classified.

CONCLUSIONThe findings in this study suggested that the oxidative stress in bodies of ACM patients was severely aggravated, and marked high oxidative constituents and low antioxidants and antioxidases in the human body might increase the relative risk of inducing acute coxsackie virus myocarditis, and measuring the values of NO in plasma, SOD and LPO in RBC might increase the correct rates of discriminatory analysis of the ACM.

Adolescent ; Adult ; Antioxidants ; analysis ; pharmacology ; Case-Control Studies ; Coxsackievirus Infections ; physiopathology ; Female ; Free Radicals ; Humans ; Lipid Peroxidation ; Male ; Middle Aged ; Myocarditis ; pathology ; virology ; Oxidative Stress