Hepatitis C virus ( HCV) is a leading cause of chronic hepatitis, cirrhosis, and hepatocellular carcino-ma in humans.Due to the lack of suitable experimental animal models for HCV infection, the development of more effective treatment of HCV infection and vaccines has been delayed.Chimpanzee is the best experimental animal model for the re-search of hepatitis C virus ( HCV) infection.However, because of its limited in resource, expensive in breeding, and difference in clinical symptoms, thus developing new experimental animal models for HCV-related basic and applied re-search is imminent.In recent years, as a surrogate animal model, the development of rodent model and other models has been achieved a lot of progress.Using such as genetically modified experimental techniques, those surrogate animals were infected with HCV in vivo and were successfully applied to research in several areas.In this review, we will focus on the a-chieved progress in naturally infected animal model and transgenic surrogate experimental models, and their advantage and limitation in usage in study on the pathogenetic mechanism of infection, drug evaluation and development of vaccines, and will also discuss the future direction of development of experimental animal models for research of infection with HCV.

Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.

This study is to investigate the effects of cisplatin combined with heparanase inhibitor OGT2115 on proliferation, invasion and migration of human nasopharyngeal carcinoma cells CNE-2 and to provide a new target for the treatment of metastasis of nasopharyngeal carcinoma. MTT assay was used to detect the cell viability of CNE-2 after exposure to different concentrations of DDP (2, 4, 8, 16 and 32 micromol x L(-1)), different concentrations of OGT2115 (0.4, 0.8, 1.6, 3.2 and 6.4 micromol x L(-1)), and DDP combined with OGT2115. Transwell assay was applied to analyze the effects of drugs on invasion and migration of human nasopharyngeal carcinoma cells. Wound healing assay was performed to detect cell migration and heparanase activity was analyzed by ELISA. MTT results showed that DDP can inhibit the proliferation of CNE-2 cells in a time and concentration-dependent manner, with an IC50 of 24.03 micromol x L(-1) at 24 h (P < 0.05), low concentration of DDP has almost no inhibitory effect on cell invasion and migration. DDP combined with OGT2115 can significantly inhibit cell invasion and migration. Inhibition of heparanase can significantly enhance anti-invasion and anti-proliferation of DDP.

This study was aimed to explore the effect of Periplaneta Americana against alcohol-induced liver injury in mice, in order to provide a theoretical basis for the industrial production of P. Americana. Kunming mice were randomly divided into six groups, which were the normal group, model group, positive group, high-, middle-, low-dose of P. Americana groups. Intragastric administration of Tiopronin Enteric-coated Tablets 100 mg·kg-1 was given to the positive group. Intragastric administrations of whole powder of medicine were given to the high-, middle-, low-dose groups with the dosage of 6.667, 3.333, 1.667 g·kg-1, respectively. The drugs were given daily for 10 con-secutive days. After 3h of the 10th day drug administration, intragastric administration of distilled water was given to the normal group, while 14mL·kg-1 of 56℃ Red Star Liquor was given to other groups. No food was given but water for 12h. Blood was collected from the orbit. The ALT, AST and GGT in blood serum of mice were measured. The liver was dissected and liver coefficient was calculated. Histopathological examination was given on liver tissues. The results showed that compared with the normal group, the level of ALT, AST in blood serum of the model group had obvious enhanced (P< 0.01), the level of GGT had obvious enhanced (P< 0.05). Compared with the model group, the level of GGT, AST of the high-, middle-dose group had obvious enhanced (P< 0.05), the level of ALT activity had obvious enhanced (P< 0.01). There were severe liver histopathological damages in mice of the high-, middle-, low-dose group. It was concluded that P. Americana had some side effects in the treatment of alcohol-induced liver injury.

Pigment epithelium-derived factor (PEDF) is an antiangiogenic factor which is effective in tumour inhibition in a variety of tumours and has not yet been studied in bladder tumour before. In this study the expression of PEDF, interleukin-1α (IL-1α) and -8 (IL-8) in bladder tumours was investigated. Immunohistochemistry was performed on 64 bladder tumour and 23 normal uroepithelium samples. Expression change of the factors was compared with clinicopathological parameters. Correlations between PEDF, IL-1α and IL-8 were analyzed. None of the factors was in relation to gender, tumour occurrence, and size or onset pattern. PEDF (P=0.014) and IL-1α (P=0.049) expression was down-regulated with grade progression. PEDF expression was lower in normal uroepithelium than in papillary urothelial neoplasm of low malignant potential (PUNLMP) (P=0.000) and carcinoma (P=0.009) whilst IL-1α (P=0.000 and P=0.000 respectively) and IL-8 (P=0.000 and P=0.023 respectively) expression was higher in the same grouping. PEDF expression had a negative correlation with IL-8 in PUNLMP (P=0.049, r=-0.578) as well as in tumour grouping (P=0.033, r=-0.276). Deranged expressional change of PEDF, IL-1α and IL-8 could be in relation to loss of differentiation from normal uroepithelium to papillary lesion and eventually to carcinoma.

Objective:To analyze the actual and expected incentives and key issues of telemedicine providers in China and to provide important references for optimizing telemedicine incentive policies.Methods:The sample data came from a questionnaire survey of 1 084 providers in 7 provinces of Fujian, Hainan, Henan, Hunan, Guizhou, Sichuan and Qinghai in October and November 2019 and a semi-structured interview data at 6 hospitals in Henan province in January 2020. Descriptive analysis was used to sort such data into text data for classified summation.Results:The greatest gaps were found between actual incentives and expected ones in expense subsidies, title promotion and excellence awards, with such gaps as high as 18.65, 28.26, 29.28 percentages among providers of junior, deputy senior and senior academic titles respectively. Hospitals fall short in providing adequate incentives for telemedicine providers, not to mention lack of differentiation and diversification in personnel incentives, and poor implementation of incentive policies.Conclusions:It is suggested to strengthen incentive demand analysis, break the homogenization bottleneck of incentive policies, and consolidate internal publicity and implementation of incentive policies.

Objective: To investigate the effects of two different sorbents(Carbon perfusion apparatus and Resin perfusion apparatus)in Double plasma molecular absorb syetem for liver failure treatment. Methods: A total of 152 cases with liver failure who were admitted to The Sixth People's Hospital of Zhengzhou, from June 2016 to May 2018 were selected and divided into DPMARS Carbon group (77 cases) and Resin group (75 cases). The two groups were observed in terms of liver function, prothrombin activity(PTA),Plasma albumin ,tumor necrosis factor alpha and interleukin-6 were detected and compared between the two groups before and after treatment. Results: ①The clinical symptoms improved in different degree in two groups, the recovery rate of Carbon cans Carbon perfusion apparatus group and Resin group separately were89.6% (69/77)、90.7% (68/75)(χ² = 0.048, P = 0.975), there were no statistical differences. There were no statistical differences between the two groups in untoward reactions(χ² = 0.235, P = 0.995), ②Compared with before treatment, TBil(t = 3.735, 3.728; P = 0.000, 0.000)、ALT(t = 5.117, 5.203; P = 0.000, 0.000)、TNF-α (t = 3.158, 3.094; P = 0.000, 0.002)、IL-6(t = 3.647, 3.559; P = 0.002, 0.003)decreased and ALB (t = 2.300, 3.065; P = 0.024, 0.003) increased significantly after treatment in both groups, and there were statistical differences. There were no signifiant differences in the changes in ALB(t = 0.316, 0.209; P = 0.657, 0.720) and PTA(t = 0.810, 0.843; P = 0.429, 0.516). ③After treatment, there were no signifiant differences in the changes in TBil、ALT、ALB、PTA、TNF-α、IL-6(t = 0.377、0.904、-1.133、-1.552、0.841、0.401; P = 0.952、0.283、0.826、0.094、0.154、0.457). Conclusion Double plasma molecular absorb syetem is effective in treating liver failure. Carbon perfusion apparatus or Resin perfusion apparatus can be combined with Specific bilirubin adsorption column for DPMARS in clinical treatment,and their effects are similar.

Objective: To understand potential categories of lifestyle behaviors and its association with traditional Chinese medicine (TCM) constitution among higher vocational medical students, so as to provide reference for the physical health management of them. Methods: From October to December 2022, 2 720 medical students from three higher vocational medical colleges in Anhui Province were selected by stratified cluster sampling method and were investigated with a questionnaire of lifestyle and TCM body constitution. Potential category analysis (LCA) and multiple linear regression were used to explore the relationship between potential categories of lifestyle behaviors and TCM constitutions. Results: The lifestyle behavior of vocational medical students were classified into three potential categories:general health group (83.60%), smoking and drinking group (4.85%) and diet preference group (11.55%). There were significant differences in the distribution of TCM constitution types among the three groups ( χ 2=46.32, P < 0.01 ). The proportion of general health group was the higher in balanced constitution (46.39%), and the proportion of smoking and drinking type was higher in phlegm dampness (25.00%). The dietary preferences were higher in phlegm dampness (20.38%), Yin deficiency (15.92%) and damp heat (20.70%). Taking balanced constitution as a reference, the occurrence risk of phlegm and dampness in smoking and drinking type and diet preference type was 1.75 times higher (95% CI =1.09-2.77) and 1.78 times higher (95% CI = 1.26-2.49) than that in general health group ( P <0.05). Conclusion There are certain aggregation characteristics in lifestyle behaviors among higher vocational medical students, which show significant relations with TCM constitutions. Targeted intervention should be carried out according to the demographic characteristics of higher vocational medical students, potential categories of behavior and lifestyle characteristics and TCM constitutions.

OBJECTIVETo study the effects of tunicamycin (a glycosylation inhibitor) combined with cisplatin on the proliferation and apoptosis of human nasopharyngeal carcinoma cells and explore the molecular mechanism.

METHODSNasopharyngeal carcinoma CNE-1 and CNE-2 cells cultured in vitro were treated with different concentrations of tunicamycin with or without cisplatin. The inhibition of cell proliferation was examined using MTT assay and colony formation assay, and the cell apoptosis was analyzed using flow cytometry with propidium iodide staining. The expressions of Bax, Bcl-2, and GRP78 in cells treated with 3 µmol/L tunicamycin with or without 6.00 µmol/L cisplatin were measured with Western blotting.

RESULTSTreatment with tunicamycin or cisplatin obviously inhibited the proliferation of CNE-1 and CNE-2 cells. Treatment with 3 µmol/L tunicamycin for 24, 36 and 48 h resulted in a viability of 72.13%, 51.97%, and 37.56% in CNE-1 cells and 85.61%, 56.95%, and 43.66% in CNE-2 cells, respectively, and the combined treatment with 6 µmol/L cisplatin lowered the cell viability to 67.97%, 47.76%, and 34.68% in CNE-1 cells and 56.89%, 37.05%, and 29.30% in CNE-2 cells, respectively. Tunicamycin at 0.3 µmol/L combined with 0.6 µmol/L cisplatin showed an obviously enhanced inhibitory effect on colony formation of CNE-1 and CNE-2 cells. Tunicamycin treatment (3 µmol/L) of CNE-1 and CNE-2 cells for 48 h induced an apoptosis rate of only 8.89% and 8.67%, but when combined with 6 µmol/L cisplatin, the cell apoptosis rate increased to 37.02% and 32.25%, significantly higher than that in cells with cisplatin treatment alone (7.25% and 6.36%, respectively). Compared with tunicamycin and cisplatin alone, the combined treatment significantly increased Bax expression and decreased Bcl-2 expression in the cells; tunicamycin up-regulated the expression of GRP-78 and enhanced the activity of caspase-3.

CONCLUSIONTunicamycin can inhibit the proliferation of CNE-1 and CNE-2 cells and enhance cisplatin-induced cell death, the mechanism of which may involve excessive endoplasmic reticulum stress response and increased activity of caspase-3.

Apoptosis ; drug effects ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Endoplasmic Reticulum Stress ; drug effects ; Heat-Shock Proteins ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tunicamycin ; pharmacology ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the effect of small interfering RNA-mediated glucose-regulated protein 78 (GRP78) knockdown on the chemosensitivity of breast cancer cells to cisplatin.

METHODSHuman breast cancer cell line MDA-MB-231 was exposed to different doses of cisplatin (0, 1, 2, 4, 8, and 16 µmol/L), and the changes in cell viability were detected using MTT assay. PI/Annexin V staining was used to observe the apoptosis of the cells in response to transfection with a small interfering RNA targeting GRP78 (pSH1Si-GRP78). Western blotting was employed to detect GRP78 expression in pSH1Si- GRP78-transfected cells after exposure to 8 µmol/L cisplatin for 24, 48 and 72 h.

RESULTSExposure of the cells to 8 µmol/L cisplatin for 24, 48 and 72 h resulted in a cell survival rate of 83.13%, 54.22% and 35.79%, respectively, but the cell apoptosis rate was only 10.8% at 24 h. Transfection of MDA-MB-231 cells with pSH1Si-GRP78 caused a cell apoptosis rate of 24.6%, which increased to 48.9% in cells with both pSH1Si-GRP78 transfection and cisplatin exposure. Cisplatin exposure caused an initial up-regulation followed then by a down-regulation of GRP78 expression in MDA-MB-231 cells, while pSH1Si-GRP78 transfection produced an obvious down-regulation of GRP78 expression.

CONCLUSIONSInhibition of GRP78 expression increases the apoptosis and enhance cisplatin chemosensitivity of breast cancer cells in vitro, suggesting the value of GRP78 as a potential therapeutic target in the clinical treatment of breast cancer.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; Cisplatin ; pharmacology ; Female ; Gene Knockdown Techniques ; Heat-Shock Proteins ; metabolism ; Humans ; RNA, Small Interfering ; Transfection