Objective To investigate the influence of knocking down Ezrin expression in combination with HSP70-induced immune killing on the apoptosis and proliferation of mouse osteosarcoma cells.Methods Human osteosarcoma cell line MG63 was cultured,the HSP70 and Ezrin-shRNA DNA fragments were cloned into the expression vector pGFP-V-RS containing CMV and pU6 promoters,and constructed the expression vector pGFP-V-RS-shRNA and pGFP-V-RS-shRNA-HSP70.The vectors were transfected into MG63 cell line,respectively.The status of transfected MG63 cells was observed by fluorescent microscope.The expressions of Ezrin and HSP70 were examined by real time RT-PCR and Western blot.Flow cytometry 、MTS test was used to detect the changes of cell apoptosis and proliferation,and changes of the expression of apoptosis and cell cycle related proteins was detected by Western blot.The specific cytotoxic T lymphocytes (CTLs) were induced by HSP70,and its killing effect on target MG63 tumor cells was analyzed by MTT assay.Results The specific vector simultaneously knocking down Ezrin and overexpressing HSP70 was constructed for the first time in China and confirmed by fluorescence microscope and gene/protein expression analysis.Compared to Ezrin knock-down alone,simultaneous HSP70 overexpression partially recovered the promoted cellular apoptosis and proliferation suppression by Ezrin knock-down,however,when compared to the normal control,the apoptosis rate of LM 8 cells was still significantly increased from 11.01％±0.22％ to 24.28％±0.50％,while the proliferation rate decreased from 395.14％±2.24％ to 310％± 2.83％.In addition,Western detected that Ezrin-shRNA could promote the expression of Bax.However,the expression of Ezrin-shRNA could reduce the Bcl-2 and Cyclin D1.Ezrin-shRNA/HSP70 also has the same effect.MTT assays revealed that the CTL cytotoxic effect on target MG63 tumor cells at all concentrations were significantly higher in CT+IL-2+HSP70 group compared with that in CT+ IL-2 group,with a cytotoxicity as high as 56.33％± 1.95％.Conclusion Simultaneous knocking down Ezrin and overexpressing HSP70 promotes the apoptosis and inhibits the proliferation of osteosarcoma cell.And the HSP70 can induce CTL which enhances the killing effect on tumor cell.And the HSP70 can induce CTL which enhance the killing effect on tumor cell.

Comparison of rCBF SPECT imaging and clinical visual function of 12 cases with complete homonymous hemianopia (CHH) were taken before and after a course of oriented dynamic color photic stimulation (ODCPS ). It was suggested that ODCPS in patients with CHH was an effective met hod for increasing visual field and improving visual function. Cerebral metabolic patterns of increasing rCBF reflected the mechanism of ODCPS effecting the patients with CHH. The rgtinal- midbrain-occipital visual path way may play an important role in mediating the increase of visual field and restoration of visual function.

Objective To compare the rate of blood transfusion and hospitalization duration between the occlusion of ventricular septal defect through transesophageal echocardiography and the repair of ventricular septal defect.Methods A total of 131 consecutive patients were included in the study from July of 2011 to December of 2011,75 patients of them received the occlusion of ventricular septal defect through transesophageal echocardiography,and 56 patients of them received the repair of ventricular septal defect.Results All of 131 patients had discharged without death.The group of receiving the repair of ventricular septal defect presented the higher rate of blood transfusion(x2 =81.83,P ＜ 0.01) and longer hospitalization duration than the other(t =46.57,P ＜ 0.01).Conclusions The group of receiving the occlusion of ventricular septal defect through transesophageal echocardiography had the lower rate of blood transfusion and shorter hospitalization duration.

Objective To evaluate clinical value of denaturing high performance liquid chromatography (DHPLC) used in detecting transforming growth factor beta receptor 3 (TGFBR-3) exons 11 and 12 polymorphism in women with idiopathic premature ovarian failure (POF).Methods From Feb.2009 to Dec.2011,110 patients with idiopathic POF undergoing treatment at Shenzhen Maternal & Child Health Institute affiliated to Southern Medical University were enrolled as POF group in this study.In the mean time,110 women under 40 years old with normal hormonal level and menstrual cycles as control group.The exons 11 and 12 of TGFBR-3 gene polymorphism were screened by using DHPLC,and results of DNA sequencing was as golden standard.Some related indexes were calculated,such as sensitivity,specificity,false negative value,false positive value,Youden index,positive predictive value,and negative predictive value.At the same time,20％ of the tested specimens were chosen randomly and detected by DHPLC again.The value of Kappa index were calculated by comparing the results between the first and second DHPLC analysis.Results The exon 11 of TGFBR-3 were not identified gene polymorphism and two nucleotide polymorphisms were identified in exon 12.For 2022 T/C polymorphism,the frequencies of CC with 0.9％ (1/110),TC with 22.7％ (25/110),TT with 76.4％ (84/110),Cwith12.3％ (27/220) and T with 87.7％ (193/220) in POF group were significantly different from CC with 0,TC with 9.1％ (10/110)and TT with 90.9％ (100/110),C with 4.5％ (10/220) and T with 95.5％ (210/220) in control group (all P ＜ 0.05).Allelic and genotypic frequencies of 2161-75 C/T were not differed significantly between the two groups (all P ＞ 0.05).As DNA sequencing as golden standard,DHPLC showed that the sensitivity was 100％,specificity was 97.9％,Youden index was 97.9％,positive predictive value was 96.3％,negative predictive value was 100％,and Kappa index was 0.888 (P ＜ 0.05).Conclusion DHPLC analysis is higher validity,reliability and practicability method in detecting TGFBR-3 polymorphism in idiopathic premature ovarian failure.

OBJECTIVETo investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C to IFN treatment.

METHODSThe HCV RNAs in PBMC were detected at the end of treatment, and 24 week and 1 year follow up after end treatment, in 16 patients who acquired complete response to IFN in 12 weeks of 24 weeks therapy.

RESULTS9 patients were HCV RNA positive in their PBMC at the end of treatment, the serum HCV RNA of 8 turned positive after 24 weeks and 1 year follow-up. In 7 patients with negative HCV RNA in PBMC, only two patients relapsed in serum HCV RNA after 1 year follow-up, and others remained viral response after 3.5 years.

CONCLUSIONHCV RNA in PBMC at the end of treatment was a predictor of the durable response to antiviral therapy in chronic hepatitis C.

Adult ; Female ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; RNA, Viral ; blood ; Recombinant Proteins

OBJECTIVETo compare postoperative blood loss under different negative pressures of drainage after total hip arthroplasty for the treatment of femoral neck fractures.

METHODSFrom January 1st to December 30th 2013, 74 patients with femoral neck fractures treated with total hip arthroplasty were randomly divided into two groups: high negative pressure drainage group and low negative pressure drainage group. In high negative pressure drainage group, there were 34 cases including 10 males and 24 females, with a mean age of (75.94 ± 9.02) years old, and the patients were treated with 60 kPa negative pressure of drainage. In the low negative pressure drainage group, there were 40 cases including 13 males and 27 females, with an average age of (74.93 ± 8.90) years old, and the patients were treated with 30 kPa negative pressure of drainage. The amount of total drainage, total blood loss, and hemoglobin change were compared between these two groups.

RESULTSAll the patients got primary healing without infections. In high negative pressure drainage group,the change of hemoglobin was (41.74 ± 15.69) g/L, total blood loss was (1,217.73 ± 459.50) ml and the drainage volume was (312.94 ± 103.44) ml; while in low negative pressure drainage group,the results were (34.90 ± 12.90) g/L, (904.01 ± 381.58) ml and (129.25 ± 44.25) ml separately. All the results in high negative pressure drainage group were higher than those in the other group. Three days after operation, the change of hemoglobin was (46.00 ± 13.29) g/L and total blood loss was (1,304.72 ± 421.75) ml; while in low negative pressure drainage group, the changes of hemoglobin was (43.87 ± 11.39) g/L and total blood loss was (1,196.78 ± 344.20) ml; there were no statistically significant differences between two groups.

CONCLUSIONWhen placing drainage devices after total hip arthroplasty for the treatment of femoral neck fractures, the level of negative pressure should be chosen according to preoperative level of hemoglobin and HCT in patients. For old patients with femoral neck fracture, low negative pressure is more suitable.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Case-Control Studies ; Female ; Femoral Neck Fractures ; surgery ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Postoperative Hemorrhage ; prevention & control

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Objective To assess the influence of length of the alanine tract of forkhead box E1 (FOXE1) gene on genetic susceptibility to idiopathic premature ovarian failure (POF). Methods Totally 110 patients with idiopathic POF were recruited between February 2009 and December 2012 at the Affiliated Shenzhen City Maternity and Child Healthcare Hospital of Southern Medical University. Controls (n=110) were individuals with normal menstrual cycles, normal FSH concentrations. The polyalanine tract and flanking sequence of FOXE1 were screened using the multiplex ligation-dependent probe amplification (MLPA) technique and direct sequence technique. Results The most frequent of FOXE1 polyalanine stretch length was 14 residues in both groups. The length of FOXE1 polyalanine reported in this study varied from 12 to 16 alanines, and three variants of FOXE1-polyalanine length, containing 12, 14, or 16 alanine residues, and 5 different genotypes were identified. The most common genotypes were 14/14 homozygote, occurring with the frequency of 81.8% (90/110) in the POF group, while 96.4% (106/110) in control subjects, respectively. The incidence of 14/14 genotypes of FOXE1-polyalanine was significantly lower in patients with POF (χ2=119.730, P=0.001) in comparison to the controls. There were significantly higher frequencies of the 16/16 genotypes in cases with POF [10.0% (11/110) versus 0; χ2=3.403, P=0.001], as compared with the controls. The FOXE1 14 alanine allele was significantly less common in the POF patient group than the controls [84.5% (186/220) versus 98.2% (216/220); χ2=25.923, P=0.001]. The FOXE1 16 alanine allele was significantly more common in the POF patient group than the controls [12.7% (28/220) versus 1.8% (4/220); χ2=19.412, P=0.001]. Conclusions The polymorphism of the polyalanine tract of FOXE1 gene have a certain relevance for the genetic aetiology of idiopathic POF.

Objective To investigate the clinical effect and the prospect of Infliximab in treatment of intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients.Methods Clinical features,inflammatory markers and coronary changes were observed in 2 cases of IVIG-resistant KD patients hospitalized in Peking University First Hospital,who were treated effectively by Infliximab.Relevant researches on the mechanism and progress of the Infliximab treatment for IVIG-resistant KD in the last 10 years were reviewed at the same time.Results Two KD patients hospitalized in Peking University First Hospital had been treated with 2 g/kg IVIG for 2 times and followed by methylprednisolone treatment.However,fever and other clinical manifestations occurred again after 2 days and 6 days when temperature returned normal.They both defervesced and all the symptoms were improved after 1 dose of Infliximab (5 mg/kg) by laboratory examinations.Four published literatures of the basic research and 9 retrospective or prospective clinical researches of Infliximab treatment of KD showed that Infliximab alleviated the inflammatory level in the KD patients significantly.Complete remission was up to 72.73％-92.11％.Those KD patients defervesced within 12 h,with dramatic improvement of symptoms and signs.Arthralgia also disappeared in the patients with arthritis.Only 1 case was complicated with hepatitis in the acute phase and cholecystitis in recovery time.A phase 3 randomised,double-blinded,placebo-controlled trial had been done to assess the addition of Infliximab to the standard therapy.Conclusions Infliximab is a feasible choice for IVIG-resistant KD patients.Efficacy and safety of Infliximab for KD treatment have been proved in the literature.However,Infliximab for KD treatment has not been indicated in the drug instruction,so the informed consent from the guardians and Ethics Committee is needed.

Objective To observe the preventive effect of the new nursing mode on the formation of deep venous thrombosis(DVT)in pregnant women with high risk factors.Methods 186 pregnant women who had high risk factors were selected,by using digital table method they were randomly divided into control group(93 cases)and observation group(93 cases).The control group was treated with routine nursing,the observation group was given the new nursing mode on the basis of routine nursing.The postpartum 2 weeks of lower limb swelling,pain,lower extremity deep vein thrombosis formation of incidence,and the nursing satisfaction were compared between the two groups. Results 2 weeks postpartum,the incidence rates of lower limb pain occurrence,lower extremity deep vein thrombosis in the observation group were 8.6%,2.1%,which in the control group were 20.4%,9.7%,there were statistically significant differences between the two groups(χ2 =4.56,4.82,all P <0.05).The satisfaction of the observation group was 95.6%,which of the control group was 81.7%,the difference between the two groups was statistically sig-nificant(χ2 =5.27,P <0.01).Conclusion The new nursing model used in pregnant women with high risk factors can effectively prevent the occurrence of DVT,reduce the incidence of DVT,improve the satisfaction of nursing.