BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.

Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P＜0.05)while,the combination therapy group achieved the best results(P＜ 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)％(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P＜0.05) and the expression of Bcl-2 was higher(P＜0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P＜0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.

Background and purpose:Adenosine triphosphate-binding cassette superfamily G member 2 (ABCG2), which has been found over-expressed in a variety of cancer cells, takes part in the drug resistance of cancer through effux of anticancer drugs. The purpose of this study was to investigate the mechanisms of human glioblastoma cells sensitivity to pyropheophorbide-a methyl ester (MPPa)-mediated photodynamic therapy (PDT) eradicating tumour cells and its relationship to ABCG2.Methods:U87 and A172 glioma cell lines in the logarithmic growth phase were selected and exposed to the treatment of MPPa-PDT and MPPa-PDT+fumitremorgin C (FTC) respectively. The cell viability was measured with the use of CCK-8 assay. The expression of ABCG2 was detected by Western blot. The intracellular contents of MPPa in each group without illumination were tested by lfow cytometry. Flow cytometry with AnnexinⅤ-FITC/PI double staining was used to detect the cell apoptotic rate. DCFH-DA staining was used to assess the generation of intracellular reactive oxygen species (ROS).Results:The MPPa-mediated PDT could eradicate A172 and U87 cancer cells in an energy-dependent manner. The light energy density in A172 was 8 times of that in U87 when the cell viability reached median lethal dose after MPPa-mediated PDT. The high expression of ABCG2 in A172 cells affected the accumulation of intracellular MPPa. Inhibition of ABCG2, not only could enhance the eradicating effect of MPPa-PDT on A172 cells, but also could increase the yield of ROS triggered by MPPa-PDT and the accumulation of intracellular MPPa.Conclusion:The human glioblastoma cell line A172 is insensitive to MPPa-mediated PDT. The mechanism may relate to ABCG2, which decreases the MPPa content in cancer cells through effux of MPPa, resulting in decline of cytotoxicity.

Protein-protein interactions between viruses and hosts are common during viral infection and replication. In this study, a cDNA library from larvae of Plutella xylostella was constructed and used for screening of genes encoding proteins interacting with Plutella xylostella granulovirus (PlxyGV) proteins. Two cDNA clones containing genes encoding proteins interacting with PlxyGV PP31 were identified by yeast two-hybrid assays. Sequence analysis showed that the genes encoded homologues of receptor for activated protein C kinase (RACK) and methionine aminopeptidase 2 (MetAP2), respectively. The P. xylostella rack gene and the PlxyGV pp31 was expressed in an E. coli strain to produce proteins fused with a 6-His or a GST tag. It was shown that the rack was expressed as a 38kD peptide as prospected. The 38kD His-tagged peptide was co-purified with GST-PP31 by GST-bind resin in GST-pulldown assays, confirming interaction between the PlxyGV PP31 and the RACK protein of P. xylostella.
Aminopeptidases ; genetics ; physiology ; Animals ; Gene Library ; Granulovirus ; physiology ; Metalloendopeptidases ; genetics ; physiology ; Moths ; virology ; Receptors for Activated C Kinase ; Receptors, Cell Surface ; genetics ; physiology

With improved overall survival of cervical cancer patients, the importance of the quality of life (QOL) is increasingly recognized. This study was conducted to compare the QOL of women with different stage cervical cancer before and after treatment to facilitate improved cervical cancer prevention and treatment. We used the generic Medical Outcomes Study Short Form-36 (MOS SF-36) to collect QOL information. Based on SF-36, we interviewed cervical cancer patients at West China Second Affiliated Hospital and Sichuan Cancer Hospital between May 2010 and January 2011. A total of 92 patients with precancerous lesions, 93 with early cancer, and 35 with advanced cancer responded to our survey. Average physical component summary (PCS) scores were significantly different between the three groups at every time point (P < 0.05). Average mental component summary (MCS) scores were significantly different between the three groups after treatment (P < 0.05). Average PCS and MCS scores increased gradually from the pretreatment to posttreatment period for patients with precancerous lesions. However, they reached the lowest at 1 month after treatment for patients with early and advanced cancers and rebounded between 1 and 6 months after treatment. Our results indicate that patients with precancerous lesions and early cervical cancer show better overall QOL than do those with advanced cervical cancer. Additionally, patients with early cancer recover more quickly than do those with advanced cancer in terms of both physical and mental functions. Thus, early detection and treatment initiatives may improve the QOL for patients with precancerous lesions and cervical cancer.
Adult ; Aged ; Carcinoma in Situ ; pathology ; therapy ; Cervical Intraepithelial Neoplasia ; pathology ; therapy ; Chemoradiotherapy ; China ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Lymph Node Excision ; Middle Aged ; Neoplasm Staging ; Precancerous Conditions ; pathology ; therapy ; Quality of Life ; Surveys and Questionnaires ; Uterine Cervical Neoplasms ; pathology ; therapy ; Young Adult

Objective To develop a method for the simultaneous determination of 15mycotoxins in peanuts by ultra high performance liquid chromatography-tandem mass spectrometry with QuEChERS EMR-Lipid approach and stable isotope dilution. Methods The samples were extracted by 2% formic acid acetonitrile-water (50 : 50, V/V) and then purified with QuEChERS EMR-Lipid approach.The mycotoxins were fully separated on a pentafluorophenyl column under a gradient elution with methonal-0.01%formic acid aqueous solution.The mycotoxins were analyzed by UPLC-MS/MS with multiple reaction monitoring (MRM) mode and quantified by isotope internal standard method. Results Fifteen mycotoxins had good linear relationship in the certain correlation ranges with the correlation coefficients all above 0.995 and the detection limits were 0.1－10 μg/kg.The mean recoveries ranged from 81.2% to 115.3% with RSD (n=6) varying from 2.1% to 10.7%. Conclusion The method is simple, highly sensitive, practical, and proves to be suitable for quantitative analysis of 15 mycotoxins in peanuts.

Objective To make uncertainty evaluation of total arsenic in pork by using the national food safety standard "GB 5009.11-2014 determination of total arsenic and inorganic arsenic in food", and discuss the influence of each uncertainty component on the determination results so as to improve the accuracy of the experimental results. Methods The uncertainty sources that affected the measurement results in the measurement process, including repeatability measurement, sample weighing, sample dilution, standard concentration, standard curve and so on were evaluated.The uncertainty introduced by the repeatability measurement was evaluated by class A, the uncertainty caused by other factors was evaluated by class B, and the phase was calculated according to each component.The synthetic standard uncertainty and the relative expanded uncertainty were discussed, and so were the influence of the uncertainty components on the measurement results. Results According to the contribution of uncertainty from large to small was the sample standard concentration, standard curve, repeatability determination, sample weighing and sample dilution.The calculated relative composite standard uncertainty was 0.0405, and the relative expanded uncertainty was 0.081 (k=2). Conclusion The experimental results show that the concentration of standard substance has a great influence on the measurement process, so it is necessary to standardize the dilution process and simplify the dilution procedure to improve the accuracy of measurement results.

Objective To evaluate the value of transesophageal echocardiography (TEE) in transthoracic minimally invasive device closure of ventricular septal defect(VSD).Methods A total of 164 cases of VSD were recruited as candidates to receive transthoracic minimally invasive device closure between January 2007 and October 2010,including 138 perimembranous VSDs,3 muscular VSDs and 23 supracristal VSDs.Among these groups,85 male patients were included.Four-champer view,five-champer view,left ventricular long-axis view,short-axis view and right ventricular inflow view were detected to evaluate the availability of device closure.Results A total of 152 cases(92.7%)were successfully closed with a device.All the patients were followed up more than 3 months arranged with a standard protocol.No complete atrioventricular block (CAVB) or associated valvular complications were observed.Three of the five cases with traced residual shunt after device closure closed spontaneously.Conclusion TEE plays an important role in transthoracic minimally invasive device closure of ventricular septal defect,which has been proved by the good follow-up results without CAVB and associated valvular complications.

OBJECTIVETo review the long-term clinical effect of composite transplantation of allogeneic acellular dermal matrix (ADM) and split thickness skin autograft (STSG).

METHODSNineteen patients with 34 wounds transplanted with allogeneic ADM combined with STSG who were hospitalized from March 2001 to October 2008 were enrolled as composite transplantation group (CT). Another 9 patients with 11 wounds transplanted with STSG admitted within the same time frame were enrolled as control group (C). All patients were followed up for longer than 2 years. Color, evenness, texture, contracture, sensation, and complications of transplanted skin were assessed using a modified Manchester Scar Scale (1-4 scores, the higher the score, the poorer the situation). The scar formation on skin donor sites was assessed by the Vancouver Scar Scale. Patients' degree of satisfaction and health status during the transplantation period were investigated in the form of questionnaire. The skin tissue structure of 4 patients was observed with histological method. The joint range of motion was assessed by the neutral position before and after operation and at follow-up. Data were processed with nonparametric test, chi-square test or t test.

RESULTS(1) The evenness, contracture, and texture of transplanted skin in CT group scored (1.6 ± 0.5), (1.8 ± 0.8), and (1.5 ± 0.8), respectively, which were significantly lower than those in C group [(2.0 ± 0.7), (2.2 ± 0.9), and (2.3 ± 0.7), with Z value respectively -2.058, -2.220, -2.323, P values all below 0.05]. Scores of color, sensation, and complications of transplanted skin in two groups were close to each other (with Z value respectively -0.628, -0.428, -2.520, P values all above 0.05). (2) Mild scar formation was observed in one of the skin donor sites in CT group. (3) Information as obtained from questionnaire showed no statistical difference between two groups in pinching, itching, and satisfaction degree (with χ(2) value respectively 0.187, 0.019, 2.628, P values all above 0.05). (4) Nerve fibers were seen in hand tissue 2 years after operation. ADM did not induce severe inflammatory responses in the site of grafting. (5) Eleven joints in CT group recovered or improved in function; while the other two joints required secondary surgery. Obvious contracture was observed in the two joints in C group.

CONCLUSIONSAllogeneic ADM combined with STSG transplantation prevents scar contracture and has obvious effect in improving function and appearance. There is no problem in regard to safety for its existence in either adult or children.

Adolescent ; Adult ; Burns ; surgery ; Child ; Child, Preschool ; Dermis ; transplantation ; Female ; Follow-Up Studies ; Humans ; Male ; Skin Transplantation ; methods ; Skin, Artificial ; Time ; Transplantation, Autologous ; Transplantation, Homologous ; Young Adult

Clinical pathway is an important quality management tool for regulating medical behavior both at home and abroad, and an important means of controlling medical costs in the reform of medical insurance payment methods.The author reviewed the current development status of clinical pathways both at home and abroad, focusing on summarizing the development experience of foreign countries, and analyzing the shortcomings in the development of clinical pathways in China from the perspectives of formulation, implementation, and evaluation. It is proposed that China should establish and improve the regulatory and incentive mechanisms for clinical pathways, accelerate the construction of supporting medical security systems, explore new incentive transmission models, attach importance to the role of patient participation in the formulation and implementation of clinical pathways, and so on, in order to provide reference for promoting the efficient development of clinical pathways in China.