Triple negative breast cancer (TNBC)is difficult to benefit from endocrine therapy or tras-tuzumab targeted drug therapy.Biological overexpression of breast cancer susceptibility gene 1,p53 gene,vas-cular endothelial growth factor and microRNA suggests that TNBC is easy to metastasis and recurrence and has a poor prognosis.Exploring the molecular subtypes of TNBC,setting out the treatment plan for subtypes and finding the corresponding monoclonal antibody targets are the research direction of TNBC in the future.

Gene therapy has emerged as an efficient modality to treat human diseases.This method is based on the transfer of genetic material to tissues to induce a curative effect.Gene therapy vectors are molecular constructs used to facilitate the penetration of genomic sequences inside the cells.Viral vectots have however several limitations when administered directly to the patient.They may cause significant toxicity by activating innate immunity or by eliciting an adaptive immune response against viral proteins.In addition,targeting the vector to the desired site is an issue when given systemically.The use of cells as vehicles for gene therapy vectors has many advantages.The combination of cell-viro-gene therapy has been thought as a new and promising strategy for therapy of cancer.The targeting vector to cancer stem cells will become a new direction in the field of gene therapy.In this article,we will introduce progressions,limitations and future directions of gene therapy of liver cancer.

Fresh hepatocarcinoma tissue and spleen samples were taken from patients during surgery. B cells from spleen were purified and activated. The hepatocarcinoma vaccine was made by cell fusion between hepatic tumor cells and activated B cells. PEG was used as the fusion agent. The fusion cells were cultured and deactivated. MHC Ⅱ and B7 molecules on activated B cells were determined by flow cytometry. Fusion rate and recovery rate of cells after refrigeration were determined respectively at the same time. The results showed that MHC Ⅱ and B7 molecules on the activated B cells were enhanced comparing with B cell. The fusion rates of three cases were 66 84%, 74 43%, and 76 55%, respectively. The recovery rates of cells were 95% and 97% after DMSO and glycerol refrigeration, respectively. The results suggest that the hepatocarcinoma vaccine owns high fusion rate and recovery rate of cells after refrigeration. It's easy to make and store. So the hepatocarcinoma vaccine is suitable for clinical use.

Objective To study complement receptor typeⅠ(CR1)activity of erythrocytes in pa-tients with SLE.Methods Using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP),CR1rosette(RBC-CR1R)and immuno-complex rosette(RBC-ICR)on red cell,the ery-throcyte complement receptor I type(ECR1)genomic density polymorphism(HH type,HL type,LL type)and erythrocyte CR1immune activity were determined in32patients with SLE and in48normal individuals.Results It was found that HH type rate of ECR1density polymorphism in patients with active SLE was significantly lower(10/16,62.5%)than that(13/16,81.3%)in patients with stable SLE.The level of CR1immune activity in HH type was significantly higher than that in HL,LL type of SLE,and significantly dif-ferent from that in48normal individuals(P

The purpose of this study is to investigate the effects of multiple-trough sampling design and nonlinear mixed effect modeling (NONMEM) algorithm on the estimation of population and individual pharmacokinetic parameters. Oxcarbazepine and tacrolimus were used as one-compartment and two-compartment model drugs, respectively. Seven sampling designs were investigated using various number of trough concentrations per individual ranging from 1-4. Monte Carlo simulations were performed to produce state-steady trough concentrations. One-compartment model was used to fit simulated data from oxcarbazepine and tacrolimus. The accuracy and precision of the estimated parameters were evaluated using the median prediction error (PE), the median absolute PE and boxplot. The results indicated that trough concentrations could yield reliable estimates of apparent clearance (CL/F). For oxcarbazepine, as the number of trough concentrations per subject increased, the accuracy and precision of CL/F, between-subject variability (BSV) of CL/F and residual variability (RUV) tended to be improved. For tacrolimus, however, although no improvement were observed in the accuracy of CL/F and BSV of CL/F, the PE distribution ranges were significantly narrowed and the RUV estimates were less bias and imprecise. In terms of algorithm, Monte Carlo importance sampling (IMP) and IMP assisted by mode a posteriori estimation (IMPMAP) were consistently better than other methods. Additionally, the sampling design had no significant effects on the individual parameter estimates, which were only depended on the interaction between BSV and RUV in various algorithms. Decreased in BSV and RUV levels can improve the accuracy and precision of the estimation for both population and individual pharmacokinetic parameter estimates.

Objective To investigate the epidemiological characteristics of polyomavirus type BK infection in renal transplant recipients.Method We systematically screened for active BKV infection preoperation and at 0.5,1,3,6,9,12 and 15 months after transplantation in 116 renal transplant recipients.The screening tests included urine cytology (by the Papanicolaou method) and BKV DNA PCR (the kit for testing the BK virus) assay of both urine and plasma,and the results were recorded.Renal biopsy was performed if the graft function was deteriorated gradually or the loads of BKV replication were very high.Routine histopathological examination and immunohistochemistry were performed on renal tissues from partial patients who received the tests of renal biopsy.Result Throughout the follow-up of 15 months,urinary decoy cells (median 8/10 HPF,[1～ 48/10 HPF]),BKV viruria (median 2.63 × 105 copies/mL,[1.78 × 103 ～ 8.54 × 109 copies/mL]),BKV viremia (median 2.70 × 104 copies/mL,[1.95 × 103 ～6.31 × 106 copies/mL]),and BKVAN (4 patients) occurred in 53.46％,24.17％,20.72％ and 3.45％ of renal-transplant recipients,respectively.The positive rate of the decoy cell and BKV DNA in urine reached the peak at the third month to the ninth month after transplantation,and the peak time of the BK viremia was the fifth month post-transplantation throughout the follow-up period.The change in BKV DNA level remained constant in blood and urine throughout the follow-up period.Conclusion The peak time of BKV infection was apparently three to nine months after transplantation,suggesting the importance of monitoring urine cytology and BKV DNA loads in post-transplantation patients closely during this period in order to reduce BKVAN after transplantation.

Objective To compare the applied value of BK virus DNA load detection in urine and plasma for diagnosis BK virus nephropathy (BKVN) in renal transplantation recipients.Method In 88 renal transplantation recipients receiving renal allograft from February 2011 to January 2012 in our institute,BK virus DNA load in urine and plasma was detected by using real-time PCR,and renal biopsy was performed on the recipients with gradual deterioration of the graft function or the loads of BKV replication being very high.The diagnosis of BKVN was confirmed by using immunohistochemistry.Results Of 88 recipients,there were 35 cases (39.8％) of viruria,18 cases (20.5％) of viremia and 5 cases (5.7％) of BKVN.The median BKV DNA load in both urine and plasma in BKVN recipients was significantly higher than in non-BKVN recipients (P＜0.05).The viruria sensitivity and specificity for BKVN were 100％ and 57.3％ (P =0.03),and the viremia sensitivity and specificity for BKVN was 100％ and 82.9％ (P =0.0002),respectively.We regraded viral load ≧ 105 copies/mL in plasma or ≥107 copies/mL in urine as the best discriminant cut-off value to predict the disease and to identify patients at risk of developing BKVAN.The positive cut-off value of urine's positive predictive value (PPV+) was 26.3％ and negative predictive vaule (PPV-) was 95.7％,and the positive cut-off value of plasma's positive predictive value (PPV +) was 83.3％ and negative predictive vaule (PPV-) was 98.8％.Conclusion The viral load ≥105 copies/mL in plasma can be used as the best discriminant cut-off value to predict the disease and to identify patients at risk of developing BKVAN,but the cut-off value of urine should be only used for screening BKV infection.

0.05) in the maimed group, indignation score was high, depression score was low and self-esteem score was high ( P

BACKGROUND:Our previous studies have shown that salidroside can induce bone marrow mesenchymal stem cells directly into neuron-like cells, and Ca2+signal is one important way to achieve its biological signal transduction. OBJECTIVE:To investigate the role and mechanism of the calcium/calmodulin (Ca 2+/CaM) signaling pathway inducing bone marrow mesenchymal stem cells to differentiate into nerve cells. METHODS:Bone marrow mesenchymal stem cells were divided into two groups:control groups and salidroside groups. Salidroside groups were induced with different concentrations of salidroside (5, 10, 20, 50 and 100 mg/L) for 24 hours and 100 mg/L salidroside was added to culture cells for different time (12, 24, 48 and 72 hours). Western blot assay was used to detect the expression levels of neural cellmarker, microtubule-associated protein 2, and the important protein of Ca2+/CaM signaling pathway:CaM and calmodulin dependent kinase II (CaMK II). Then Ca2+/CaM signaling pathway specific blockers were applied to cells respectively for 30 minutes, including 500 μmol/L EGTA (Ca 2+chelator), 1 mmol/L Nifedipine(L-type Ca2+channel blocker) and 10 mmol/L LY294002 (PI3K inhibitor). Then, 100 mg/L salidroside was added and cultured for 24 hours. Western blot assay was used to detect the expression of neuron-specific enolase and CaM in the Ca2+/CaM signaling pathway. RESULTS AND CONCLUSION:(1) After inducing with salidroside, the expression of microtubule-associated protein 2 were upregulated (P<0.01), indicating that salidrosid can induce the neuronal differentiation of bone marrow mesenchymal stem cells. (2) After different concentrations of salidrosid induced bone marrow mesenchymal stem cells for 24 hours, the expressions of CaM and CaMK II were significantly upregulated in the 10 mg/L group ( P<0.01);For the 100 mg/L salidrosid that was added for cellinduction for different time, the expressions of CaM and CaMK II were significantly downregulated in 72-hour group (P<0.01). (3) After blocking extracellular Ca2+and PI3K signaling pathway, the expressions of neuron-specific enolase and CaM were higher than those in salidrosid groups (P<0.05). These results suggest that salidrosid can induce bone marrow mesenchymal stem cellto directly differentiate into nerve cells by inhibiting the Ca2+/CaM signaling pathway.

Objective To analyze the causes of failed transcatheter closure for ventricular septal defects (VSD)in children. Methods One thousand two hundred and eighty children aged 13 to 141 months who underwent transcatheter closure from June 2009 to September 2013 in Guangdong General Hospital were selected. There were 43 failures(3. 36% ). The clinical data including transthoracic echocardiograph( TTE),radiography,interventional ap-proach and surgical findings were analyzed. Results Forty - three patients included 25 male and 18 female. The pa-tients' ages ranged from 13 to 141(43. 0 ± 31. 9)months and their weight ranged from 10 to 35(16. 3 ± 5. 59)kg. The causes of failure including doubly committed subarterial VSD misdiagnosed as perimembranous VSD(PMVSD)or intracristal VSD were in 6 patients. The size of occluder was too small in 13 cases,and there were statistical differences between three measurements of size of VSD(F = 19. 134,P = 0. 001). The size of VSD measured by left ventricular an-giography was significantly smaller than that measured by TTE,and there was statistical difference[(4. 78 ± 1. 11) mm vs(6. 48 ± 1. 43)mm,t = 4. 50,P = 0. 001]. The dimension of VSD measured by left ventricular angiography was significantly smaller than that measured by surgical findings,and there was statistical difference[(4. 78 ± 1. 11) mm vs(7. 02 ± 1. 08)mm,t = 5. 92,P = 0. 001]. But,the size of VSD measured by TTE had no significant difference compared with that measured by surgical findings(t = 1. 42,P = 0. 168). Aortic regurgitation occurred in 14 cases;atrioventricular block or left bundle branch block in 3 patients;tricuspid stenosis in 2 cases and residual shunt in 5 pa-tients. Conclusions Doubly committed subarterial VSD may be misdiagnosed as PMVSD or intracristal VSD. In the ca-ses of VSD concomitant with aortic valve prolapse,size of the occluders should be referred to VSD dimensions measured by TTE. In the cases of VSD adjacent to aortic valve,suitable occluders should be selected and operation technique should be improved to avoid aortic regurgitation.