Equipping tumor-targeting carrier with cell-penetrating peptide with high transduction efficacy has become a trend. According to the fusion modes and tumor-targeting mechanisms, cell-penetrating peptides can be divided into five categories: first, self-targeting cell-penetrating peptides; second, fusion carriers made of targeting ligands and cell-penetrating peptides; third, cell-penetrating peptide-modified nano-carrier; fourth, tumor-microenvironment-targeting cell-penetrating peptides; fifth, other special cell-penetrating peptides. Fusion carriers, which can not be effectively released into the cytoplasm after transducted into target cells, can be further modified to increase their efficacy. In all, cell-penetrating peptide introduced into tumor-targeting carrier should provide a new era for anti-tumor pharmacy research and cancer therapy.

he aim of this study was to obtain a cell-penetrating cytoglobin (Cygb), which combines the transmembrane function of cell-penetrating peptides TAT with the anti-aging and anti-fibrotic role of cytoglobin. The Cygb gene was complexed with TAT gene by overlapping PCR, inserted into the vector pET22b to construct the recombinant expression plasmid (pET22b-TAT-Cygb) and then transformed into Escherichia coli BL21 (DE3). The fusion protein TAT-Cygb, whose expression was induced by lactose, was purified by CM Sepharose Fast Flow Protocol and verified by Western blotting. The final TAT-Cygb had a molecular weight of 23 kDa with 95% purity, as shown by SDS-PAGE. As demonstrated by bioactivity experiments, TAT-Cygb exhibited a high specific peroxidase activity up to (422.30 ± 0.36) U/mg. Both TAT-Cygb and Cygb pretreatment group could protect Hacat cells against oxidation of H2O2, but only TAT-Cygb treatment group could remedy cells injuried by H2O2 (RGR = 98%), which was significantly different from Cygb treatment group (RGR = 79%). We successfully obtained the bioactive and cell-penetrating fusion protein TAT-Cygb that has the potential application in anti-aging, anti-fibrotic and anti-cancer.
Blotting, Western ; Cell Line ; Cell-Penetrating Peptides ; biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Gene Products, tat ; Genetic Vectors ; Globins ; biosynthesis ; Humans ; Hydrogen Peroxide ; Recombinant Fusion Proteins ; biosynthesis

Objective To investigate the efficiency of ring butterfly sewing in lower uterine segment aided with tourniquet in treating pernicious placenta previa combined with placenta percreta. Methods Twelve pregnant women diagnosed with pernicious placenta previa combined with placenta percreta by prenatal ultrasound in Peking University First Hospital from April 1, 2012 to November 30, 2014, were enrolled. All of them received elective cesarean delivery using ring butterfly sewing in lower uterine segment aided with tourniquet. The efficiency of this novel technique, in term of blood loss and vital signs during operation, blood transfusion, and postoperative recovery, was analyzed. Results Twelve cases were diagnosed with placenta percreta during operation, with placenta invading the perimetrium in seven cases and urinary bladder being involved in one case. Ring butterfly sewing in lower uterine segment aided with tourniquet during elective cesarean delivery was all successful in the 12 cases. The median amount of blood loss was 1 000(400-2 000) ml during operation and the amount of blood loss was lower than 1 000 ml in three cases. Two cases received no blood transfusion, and the median blood transfusion in the other 10 cases were 400(400-1 200) ml. The mean operation time was (75±22) min (43-131) min. Eleven cases had normal lochia and temperature after operation and the other one had placenta invading urinary bladder and partial placenta in situ, and developed fever at day 3 after operation, but recovered after antibiotic treatment, and no placenta tissue was seen by ultrasound at day 59 after operation. Conclusions The ring butterfly sewing in lower uterine segment aided with tourniquet is highly effective for the pernicious placenta previa combined with placenta percreta, and it is a simple, fast and fertility sparing technique with low blood loss and without usage of special equipment.

Objective To analyze the implementation effect of formative assessment in nutri-tion and food hygiene. Methods 55 students of Class One from grade 2010 majoring in preventive medicine were taken as the control group, with 56 students of Class Two as the experimental group.The control group used the summative assessment , while the experimental group used the formative assessment. Results were analyzed by course examinations and questionnaire surveys. SPSS 17.0 soft-ware was used and the measurement data were expressed by x±s. Statistical analysis was performed using t test, test, rank sum test, a Yuan linear correlation analysis, with the level of test α=0.05. Results The experimental scores and final scores of experimental group students were significantly higher than the control group and the difference was statistically significant(P values were 0.001, 0.000). The experimental group's overall result is closer to the negative skewed distribution. The ex-perimental group's average score of 89 points is significantly higher than the control group's average score of 79 points(P<0.01), and the usual results and the final results were positively correlated(r=0.865, P<0.001). The survey indicated that the students recognized the application of formative assess-ment in teaching process . Conclusion Formative assessment can help improve the teaching and learning quality in nutrition and food hygiene.

As a new kind of perioperative management strategy and ideas,enhanced recovery after surgery (ERAS) brings a series of traditional perioperative treatment measures for optimization based on evidence-based medicine.Its primary aim is to decrease surgery-related stress,complications,and also to facilitate recovery with multimodal treatments.The ERAS pathway has been widely applied and proved to be efficient in gastrointestinal,colorectal,orthopedic,thoracic and gynecological surgery.Owing to the complexity of surgical procedure,long operation time and high perioperative complication rate in liver transplantation compared with other surgeries.This new concept has not been widely accepted or recommended in liver transplantation,although some previous studies have validated its safety and effectiveness.This paper overviewed the recent literature on the specific procedures,efficacy,safety and development of ERAS applied in liver transplantation.

Objective To investigate the immunosuppressive regimen of cyclosporine A(CsA) reduced or withdrawn in the HBV-DNA positive kidney transplanted patients. Methods The program of 64 kidney transplanted patients with HBV-DNA positive from Jan,2004 to Dec,2007 were analyzed, the patients were divided into 3 groups ①CsA + MMF group(A group) ;②FK506 + MMF group(B group) ;③low dose of CsA + SRL group(C group). All the patients received entecavir to resist HBV replication and were followed up for acute rejection incidence,liverfunc- tion and HBV-DNA test for 6 months. Results There was no significant difference in 3 groups about acute rejection incidence rate. Liver dysfunction took place in 12 patients of A group (80%) ,8 patients(53%) in A group HBV-DNA became negative; 5 patients (20%) in B group appeared the liver dysfunction, HBV-DNA became negative in 18 patients(75%). 4 patients in C group(16%) appeared liver dysfunction sHBV-DNA was negative in 18 patients (72%) of C group. Conclusion It was safe and efficient for the immunosuppressive regimen of cyclosporin A re-duced or withdrawn in the HBV-DNA positive kidney transplanted patients,not increasing the incidence of acute re-jection and aggratating the liver injury.

Objective:To investigate RNA interference and apoptosis induced by LPS in kidney epithelial cells through silencing C5aR gene with small hairpin RNA (shRNA).Methods:We construct the eukaryotic expression vector of small hairpin RNA targeting rat C5aR gene,and transfected RK3E cell by electroporation,after G418 selection,so we got the stable cell line expressing C5aR shRNA.The experiment was designed into 3 groups:①normal control group,RK3E cells without transfection;②negative control group,RK3E cells transfected with blank vector;③ experimental group,RK3E cells transfected with C5aR shRNA.After incubation with LPS for 12 h,the ratio of apoptosis was tested by flow cytometry,the level of mRNA was tested by RT-PCR,and binding of ~(125)I-rrC5a to RK3E cells stimulated with LPS were performed to examine the expression of C5aR in RK3E cells.Results:Compared with the normal control group and negative control group,in the experimental group the ratio of apoptosis was significantly decreased(P＜0.01),and the expression of C5aR mRNA was significantly inhibited(P＜0.01),and binding of ~(125)I-rrC5a to RK3E cells was significantly decreased also.Conclusion:Hairpin shRNA targeting C5aR gene can lead to obvious gene silence in vitro and inhibit the cell apoptosis induced by LPS in kidney epithelial cell.

Objective To investigate the protective effect of silencing tumor necrosis factor-α (TNF-α) gene in Kupffer cells on liver ischemic reperfusion injury in rats. Methods We constructed the eukaryotic expression vector of small hairpin RNA targeting rat TNF-α gene. PBS, blank vector or TNF-α shRNA mixture were injected through portal vein 48 h before operation. The animals were divided into 4 groups: sham group, PBS group, blank vector group, and shRNA group. The left lateral and median lobes of the liver were rendered ischemic for 40 min resulting in a segmental (70 %) hepatic ischemia. The serum ALT and AST levels were determined 6 h after reperfusion. The mRNA expression of TNF-α in Kupffer cells, serum TNF-α level, concentration of the malondialdehyde (MDA) and superoxide dismutase(SOD) in the liver tissue were detected. Results Compared with the PBS and blank vector groups, ALT and AST levels were significantly decreased in the shRNA group after reperfusion for 6 h (P＜0. 05), TNF-α mRNA level, serum TNF-α level(56.6 ± 6. 7 pg/ml vs 87.8 ± 8. 7 pg/ml and 96.5±7.3 pg/ml,P＜0. 05), concentration of MDA in the liver tissue(93.4±13.3 nmol/mg vs 133.5±12. 4 nmol/mg and 136.7 ±13. 6 nmol/mg, P＜0.05 )were significantly decreased, and SOD activity was significantly increased(22.4±4.6 U/mg vs 12. 2±3. 1 U/mg and 11. 4±2. 9 U/mg,P＜0. 05). Conclusion RNA interfering TNF-α gene in Kupffer cells can protect the liver against ischemia/reperfusion injury.

Background and purpose:The association between metabolic syndrome (MS) and renal cell carcinoma (RCC) is still unknown. The aim of this study was to elucidate how MS correlates with the prevalence and malignancy of RCC.Methods:This study enrolled 398 RCC patients (350 clear cell RCC patients, 5 XP11.2 transloca-tion RCC patients, 16 papillary RCC patients and 27 chromophobe RCC patients), 160 normal persons, and 32 benign renal tumor patients. The metabolic status of the patients was assessed, and the link between MS and the prevalence or malignancy of RCC was calculated.Results:Clear cell RCC patients had signiifcantly higher rates of hypertension, higher body mass index (BMI) and longer waist circumference. Forty-eight percent clear cell RCC patients had MS, while the number was 33% for papillary RCC, 26% for chromophobe RCC, 0% for XP11.2, 17% for AML, and 25%for normal people. MS patients had signiifcant higher rates of having clear cell RCC than no-MS patients, however this kind of difference was not seen in other types of RCC. Clear cell RCC patients with higher Furhman grade had lower rates of MS.Conclusion:Patients with MS are more likely to develop clear cell RCC. Patients with high Furhman grade tumors have low MS rates, indicating that high grade tumor may have other originating mechanisms other than metabolic disorders.