Objective To express the recombinant protein VP1-Z, and investigate whether VLP-Z has the physiological functions like as wild-type VLP. Methods The expression plasmid pET15b-VP1-Z was introduced into competent E. coil BL21 (DF3)/pLys cells by transformation, and the expression of re-combinant protein VP1-Z was induced by incubation of the cells with IPTG. The protein was prepared as pre-viously described for wild-type VLP. The morphous of VLP-Z were observed by electron microscopy, and the physiological functions of VLP-Z were investigated by hemagglutination test and by immunofluorescence. Re-sults The purified VLP-Z composed of VP1-Z possessed hemagglutination activity and yielded a prominent band of 50×10~3 on SDS-PAGE and staining with Coomassie Brilliant Blue. The VLP-Z exhibited virus-like particles structure like as wild-type VLP with a diameter of 45-50 nm, which was slightly bigger than that of wild-type VLP(42-45 nm). In immunofluorescence test, VP1-Z was detected within the cytoplasm and nu-cleus after HeLa cells were inoculated with VLP-Z. Conclusion The physiological functions of recombinat-ed protein VLP-Z were comparable with wild-type VLP.

Objective To construct the vector pET15b-Z-VP1 by inserting the Z fragment into amino-terminal of JCV VP1.Methods The VP1 and Z fragment were amplified by PCR from plasmid pET15b and pEZZ18 respectively,and then they were linked by recombinant PCR.The Z-VP1 fragment was inserted into plasmid pET15b by restriction enzyme BamHⅠ and NcoⅠ.Results The VP1 and Z fragment were obtained by PCR and gel purification.The Z-VP1 fragment,which was linked by recombinant PCR from VP1 and Z fragment,was inserted into plasmid pET15b between BamHⅠ and NcoⅠ sites,and confirmed by enzyme digestion and DNA sequencing.The expression of VP1-Z was confirmed by Western blotting.Conclusion The plasmid pET15b-Z-VP1 has been constructed successfully by inserting Z fragment into amino-terminal of VP1.

As a nuclear transcription repressor,the functions of TGIF are complicated,not only represses target genes expression directly,but also takes part in the regulation of multiple important cellular signaling pathways,which are associated with the differentiation of cells and tissues,inflammation,metabolism and tumors.In past few years,more and more studies on the role of TGIF in tumors suggest TGIF may be a new therapy target in the diagnosis and treatment of tumours.This article mainly reviews the research progress of TGIF in some signaling pathways like TGF-β,MAPK,PI3K/AKT,and tumours like hepatocellular carcinoma,lung carcinoma and urinary tract urothelial carcinoma.

Pancreatic tuberculosis (PTB) is a rare,chronic,specific and infectious disease which is generally secondary to tuberculosis at the common sites of pancreas,and it has a high misdiagnosis rate due to the hidden onset and nonspecific symptoms of PTB.A patient with PTB was admitted to the First Affiliated Hospital of Zhengzhou University in June 2014.Before operation,the space-occupying lesions of the head of pancreas were detected by preoperative imaging examination,and the patient was regarded as with pancreatic cancer.Intraoperative exploration showed cystic duct involvement,and the granulomatous inflammation was detected by rapid pathological examination using frozen section technique,after that the patient received granuloma resection+cholecystectomy according to suspected PTB.The diagnosis of PTB was confirmed by postoperative pathological examination,and the patient received liver-protective and anti-tuberculosis treatments after discharge.

BACKGROUND: During xenogenic liver transplantation, major histocompatibility antigen can induce immunological rejection, and immunosuppressant can cause adverse effect on organism. Recently, treatment prior to transplantation induces immune tolerance, which is perspective for organ transplantation.OBJECTIVE: To investigate the correlation between thymus induction and immunological rejection during liver transplantation. METHODS: A total of 40 male SD rats of clean grade were selected as donors. Moreover, 30 male Wistar rats of clean grade and 10 male SD rats of clean grade were selected as recipients. The donor rats were divided into allogeneic gene transplantation, allotransplantation, cyclosporine, and thymus induction groups, with 10 rats in each group. The modified Kamada and improved two-cuff technique was used to establish a stable rat orthotopic liver transplantation model. The cyclosporine group was given cyclosporine (50 mg/kg) for 5 successive days. Thymus induction group was injected with major histocompatibility antigens (50 pL) for 5 successive days. Other groups were not given any interventions. Survival time of rats was recorded in each group. Pathological observation and mixed lymphocyte cultured were performed at days 3, 7, 14, 21, and 28 after transplantation. RESULTS AND CONCLUSION: Survival time was longer in the thymus induced group compared with other groups (> 60 days), damaged level was mild, local immunological rejection was reduced, and lymphocytes were decreased. The effect after liver transplantation was similar to allogeneic gene transplantation but superior to cyclosporine intervention (P < 0.05). This suggested that thymus induction relieved immunological rejection following liver transplantation.

BACKGROUND: Most patients who underwent liver transplantation would suffer acute rejection or transplanted liver failure resulted by chronic rejection, therefore, inducing specific immune tolerance via varied pathways is the ideal method to solve this problem. OBJECTIVE: To treat rat transplanted liver by injecting transforming growth factor β1 (TGF-β_1) plasmid, and to analyze the relationship between TGF-β1 and allograft rejection from gene level. METHODS: A total of 30 male, Wistar rats were served as allogenic liver donors, and 10 male, SD rats served as syngeneic donors Totally 40 male SD rats were served as liver recipients, and divided into 4 groups by order number table: ailogenic transplantation, syngeneic transplantation, ciclosporin, and ciclosporin plus TGF--β_1 groups. In each group, rat orthotopic liver transplantation model was established by modified Kamada and improved two-cuff technique. After modeling, rats were received cyclosporine 1-5 days in the cyclosporine group, or intraperitoneal injected ciclosporin for 1-5 days, combined with TGF-β_1 plasmid 0-2 days in the cyclosporine plus TGF-β_1 group. No intervention was performed in the other groups. The survival time of rats were recorded, and the pathological changes was detected at days 3, 7, 14, 21, and 28 after transplantation, then the mixed lymphocyte culture was performed. RESULTS AND CONCLUSION: The survival time of rats in syngeneic transplantation group and cyclosporine plus TGF-1,β_1 group was more than 60 days, which was obviously greater than that of allogenic transplantation and cyclosporine groups (P< 0.05). The histopathologic slide showed that there was moderate and severe acute rejection, with evident intrahepatic inflammatory cell infiltration in the allogenic transplantation and cyclosporine groups. Few rejections were observed in the syngeneic transplantatior group, which was close to the normal lever tissues. Mixed lymphocyte culture of the cyclosporine plus TGF-β_1 group was superior to the syngeneic transplantation group or cyclosporine group (P < 0.05). The results demonstrated that cyclosporine combined with local injection of TGF-β_1 plasmid can relieve post-transplant immune rejection.

BACKGROUND: A small number of mesenchymal stem cells (MSCs) present in bone marrow, which would gradually drop with age. OBJECTIVE: To verify the effect of adherent method on culture of bone marrow-derived MSCs. METHODS: Under anaesthesia, bone marrow cells were obtained from femur and tibia of rats, cultured by DMEM containing calf serum, placed in an incubator containing 5% CO_2 at 37 ℃. The culture medium was renewed after 24 hours, and remained periodical medium change with once per week. The weakly adherent cells were passaged. The cell morphology, growth curve, and the expression of cell-surface markers were identified by flow cytometry and immunocytochemical staining. RESULTS AND CONCLUSION: After 24 hours of culture, the cells could adhere to the walls with fusiform or triangle shapes, proliferated faster after 2-3 days, and presented whirlpool-like or clustering. The cells reached a logarithmic growth phase after 2 days, and into the late stationary phase after 12 days, which covered the bottle after 15 days. The cultured cells were positive to CD90 and CD54. The results verified that bone marrow-derived MSCs can be isolated by adherent method. This method is easy operation, and can maintain cell activity preferably.

Objective To determine the electrical conductivity of cerebrum, liver, lung and muscle of rats at different postmortem intervals for investigating the relationship between EC and PMI. Methods Healthy Sprague-Dawley rats were sacrificed and kept at constant temperature of 25°C. Cerebrum, lung, liver and muscle were extracted at different PMIs of immediate (0d), 1d, 2d, 3d, 4d, 5d, 6d and 7d, and their extraction liquids were prepared with ultrapure water at the ratio of 1g:10mL. EC were separately determined for different tissues and organs. The relationships between EC of different tissues and organs and PMI were analyzed and their regression functions were established. The characteristics of EC values for four tissues and organs were compared and their decomposition processes were discussed. Results EC of brain and muscle showed no significant changes within 1d, and increased rapidly during 2~7d; but EC of liver and lung started to increase within 1d and increased rapidly during 2~7d.The relationship between EC of different tissues and organs and PMI were well fitted with cubic equations and liver gained the highest coefficient (R2=0.96). Additional, the EC of four organs presented various increasing laws in different periods of PMI. Conclusion The EC of cerebrum, lung, liver and muscle of rats were well fitted with PMI and the determination of EC of cadaver tissues can be expected to become an effective method for PMI estimation in forensic practice.

Objective: To investigate the therapeutic effects of norcantharidin poloxamer 407(NCTD P407) slow released preparation on advanced primary liver cancer by local intratumoral injection. Methods: Fifty six advanced primary liver cancer patients with maximum diameter from 6 to 12 cm were randomly divided into test and control groups. Patients in test group (29 case) were treated by NCTD 407 intratumoral injection under ultrasonic and patients in control group (27 case) by PEI (percutaneous intratumoral absolute ethanol injection). Clinical therapeutic effects were observed respectively and compared. Results: Twelve months survival rates of patients in 2 groups were significantly different, being 31.0% in test group and 22.2% in control group ( P 0.05) in 2 groups. No evident poisonous and side effects were found in either test or control group. Conclusion: NTCD P407 administration exhibits therapeutic effects in relatively big advanced liver cancer because of the high concentration and prolonged action in the tumor foci, its comprehensive therapeutic effect is better than that of absolute ethanol administration. [

Abstract: Objective To evaluate the key quality control and protective performance test of Halcyon medical linear accelerator. Methods WS 674-2020Specification for Testing of Quality Control in Medical Linear Accelerator（ According to the hereinafter WS 674-2020） ， referred to and the manufacturer´s manual the performance of the first Halcyon medical linear accelerator in Results ， ， Hunan Province was tested. The results showed that all ten indicators of the accelerator including dose deviation ， ， （ ， ， ， - repeatability linearity daily stability and symmetry the results were 0.10% 0.03% 0.04% 0.50% and 100.50% 100.80% ）， - - respectively met the requirements of WS 674 2020. The results of manufacturer quality control indicators such as dose rate ， ， stability in beam gantry rotation isocenter and mechanical position accuracy megavolt image parameters and cone beam computer tomography image parameters met the requirements of the manufacturer´s regulations. Due to the special structure and ， - ： function of the accelerator it is difficult to detect the parameter required by WS 674 2020 as below the radiation leakage - ， ， ， ， outside the M zone the uniformity the indicators related to the light field the offset of the radiation beam axis the zero scale Conclusion - position of the rotating motion scale and others. It is difficult to carry out complete testing according to WS 6742020 for Halcyon medical linear accelerator and it is urgent for the state to issue relevant testing standards to standardize and strengthen the quality control testing of various accelerators.