Apoptosis of PK-15 cells induced by Foot-and-Mouth Disease Virus (FMDV) in vitro was reported in this paper. Typical cell apoptosis was detected by use of Hoechst 33258 fluorescence probe, agarose gel electrophoresis and in situ end-labeling (TUNEL). After PK-15 cells were infected by titration of 4.8 lg TCID50/mL FMDV for 32 h, apoptosis characteristics of nuclear condensation, fragmentation, accompanied by apoptotic bodies formation (Hoechst 33258 staining), 180-200 integer-fold sized pieces DNA Ladders (agarose gel electrophoresis) and strong green fluorescence dots (TUNEL) were all exhibited, and cell apoptosis was approximately 20%. In addition, the quantitative analysis of apoptosis in PK-15 cells induced by FMDV showed that apoptosis was correlated with infection of virus, and it was also time-dependent. Results indicate that FMDV can induce apoptosis of host cells and apoptosis plays an important role in the cytopathogencity effect of FMDV.

BACKGROUND: Clinical evidence have proved that heterotopic ossification is easily present in bony callus of patients with fracture combined with head injury, and the healing of bone fracture is accelerated obviously. OBJECTIVE: To study the expression and distribution of vascular endothelial growth factor (VEGF) in bony callus among the patients with fracture combined with head injury and simple fracture patients, and to investigate its clinical significance and action mechanism.DESIGN, TIME AND SETTING: Grouping controlled observation was performed in the Xijing Hospital, the Fourth Military Medical University of Chinese PLA between February 2006 and July 2007.PARTICIPANTS: The patients comprised two groups those who had fracture combined with head injury and those who had simple fracture. Each group had 50 patients. The fracture combined head injury group included 41 males and 9 females, aged 19-55 years; simple fracture group included 36 males and 14 females, aged 17-52 years. METHODS: The bony callus specimens were harvested at 7-10, 11-15, 16-20, 21-27 and 28-35 days post-injury. The content of VEGF in bony callus was measured at different phase by using immunohistochemistry methods, and the speed of fracture healing was also observed.MAIN OUTCOME MEASURES: ①X-ray results; ②immunohistochemical image analysis. RESULTS: In different phase of bone healing, the VEGF of the two groups were homogenous, the early expression of VEGF in osteoprogenitor cell, osteoblast and cartilage cell in the fracture combined with head injury group was significantly higher than those in simple fracture group. The osteoprogenitor cell proliferated, and the differentiations of osteoblast and cartilage cell were enhanced obviously. The VEGF level reached the peak at 7-10 days, and kept at the high level for 30 days and then gradually decreased. The peak of VEGF level in simple fracture group came at 11-15 days, kept in the high level for 20 days and then gradually decreased. The peak value in simple fracture group was significantly lower than the fracture combined with head injury group. There was statistical significance in the expression of VEGF between two groups by the paired t-test (P < 0.05). A large mount of osteotylus could be detected at 4 weeks after injury by X-ray in patients with fracture combined, with head injury, but at 7-9 weeks in patients of those who had simple fracture.CONCLUSION: The expression of VEGF in the bony callus in the group of fracture combined with head injury is significantly greater than that in simple fracture patients, and the expression lasts for a longer time. The proliferation and differentiation of osteogenitor call, osteoblast and cartilage cell are also significantly faster in the group of fracture combined with head injury, which might be one of the bone healing mechanisms.

OBJECTIVETo express the cloned gene glycoprotein I (gpI) of varicella-zoster virus (VZV), Beijing VZV 84-7 strain in insect cells and to purify its expression product.

METHODSThe gene coding for gpI of VZV was amplified from viral DNA by PCR and cloned into baculovirus transfer vector (pBacPAK9), and recombinant transfer vector plasmid pBacVZVgpI was obtained. The inserted gpI gene in the pBacVZVgpI was sequenced. Insect cells Sf 9 were co-transfected with the recombinant transfer vector plasmid pBacVZVgpI and wild type linear baculovirus BacPAK6 (digested with Bsu36I) DNA. The recombinant baculoviruses containing the VZV 84-7 gpI gene was isolated through several rounds of limited dilution. Recombinant protein gpI was expressed in insect cells Sf 9, postinfected with recombinant baculoviruses. The expressed recombinant gpI was purified by lectin affinity chromatography and its antigenicity and immunogenicity were investigated.

RESULTSThe gene coding for gpI of VZV was obtained by PCR and the gpI gene of pBacPAK9 was confirmed by DNA sequencing. The recombinant gpI was expressed in insect cells Sf 9, post-infected with recombinant baculovirus and identified by SDS-PAGE and western blotting, with its product in cell culture reaching the peak in 72 hours and with a molecular mass of 58 kd and 70 kd, the same as theoretical values. Results of immunoassay with cell lysates infected by recombinant baculoviruses indicated that recombinant protein expressed in insect cells had ability of eliciting specific antibodies against native VZV in mice and complement-dependent neutralizing antibodies. The purified recombinant gpI gave a product with a purity of more than 80%. ELISA and Western-blot analysis demonstrated that purified protein had specific VZV antibody-binding activity. This suggested that the recombinant gpI expressed in insect cells had the same biological characteristics as its native counterpart.

CONCLUSIONBaculovirus-insect cells could be used to express the gene of VZV gpI, which could provide a basis for quantitative analysis of VZV antigen, and preparation of its subunit vaccine.

Animals ; Cell Line ; DNA, Viral ; genetics ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; genetics ; Genetic Vectors ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; isolation & purification ; Spodoptera ; cytology ; genetics ; Viral Envelope Proteins ; genetics ; isolation & purification

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

Objective To explore the long-term effect of endemic arsenism on oxidative stress and immune function, and to provide scientific basis for prevention and treatment of the disease in the areas. Methods In 2009, Using cluster sampling and typical investigation, the cross-sectional study was completed. The patient groups and the internal control group were selected in the arsenism areas after 5 years quality improvement of drinking water(Silizhuang village, Daying village and Gucheng village in Shanyin county, Gucheng city, Shanxi province) and they were divided into mild, moderate, severe case and internal control groups, respectively. The external control group was selected in a non-arsenism area(Yangzhuang village in Heshengbu city). The Oxidative stress indicators were determined and analyzed [serum superoxide dismutase (SOD) activity was determined with xanthine oxidase method, glutathione peroxidase(GSH-Px) activity was determined with 2-thio-2-nitrobenzoic acid method, and mmuuity malondisldohyde(MDA) levels was determined with thiobarbituric acid method]. The immune function was determined and analyzed [immunoglobulin G (IgG) was determined with radioimmunoassay method, and serum lysozyme was determined with turbidimetric method]. Results A total of 252 people were surveyed, in which the external control group, the internal control group, mild, moderate and severe patient groups were 56, 57, 49,44 and 46, respectively. Serum SOD activities were (72.19 ± 11.75), (66.96 ± 12.02), (49.79±11.07), (48.54 ±10.56) and (47.68 ± 10.68)kU/L, respectively. The difference of serum SOD activities between the groups was statistically significant(F = 52.42, P ＜ 0.01 ). Serum SOD activities in the external control group were significantly higher than other groups (all P ＜ 0.05). The value in the internal control group was significantly higher than the 3patient groups (all P ＜ 0.05). There were no significant differences between the case groups (P ＞ 0.05). Serum GSH-Px activities of the five groups were (197.41 ± 38.54), (195.02 ± 31.93), (187.26 ± 28.22), (187.24 ± 25.40),(186.88 ± 21.84)U/mg, respectively, and the difference between the groups was not significant(H = 4.21, P ＞0.05). Serum MDA levels of the five groups were (4.51 ± 2.14), (5.88 ± 2.00), (6.44 ± 2.83), (5.89 ± 2.57),(5.88 ± 2.40)μ mol/L, respectively, and the difference between the groups was statistically significant(F = 3.36,P ＜ 0.05). The external control group was significantly lower than other groups(all P ＜ 0.05). No significant difference was observed between other groups(all P ＞ 0.05). Serum IgG levels were(11.16 ± 2.08), (8.15 ± 1.44), (8.77 ±2.54), (9.19 ± 1.97), (8.44 ± 2.52)g/L, respectively, and the difference between the groups was statistically significant(H = 52.92, P ＜ 0.01 ). The external control group was significantly higher than other groups(all P ＜0.05). No significant difference was observed between other groups(all P ＞ 0.05). Serum lysozyme levels were (13.57 ± 5.16), (10.05 ± 3.96), (8.78 ± 3.35), (8.72 ± 3.76), (9.38 ± 4.26)mg/L, respectively, and the difference between the groups was statistically significant (H = 35.00, P ＜ 0.01 ). The external control group was significantly higher than other groups(all P ＜ 0.05). No significant difference was observed between other groups(all P ＞ 0.05). Conclusions The effect of arsenic on the body's oxidative stress response and immune function persists after 5 years of drinking low arsenic water. In addition to intensify arsenic removal from drinking water, it should also strengthen the monitoring of population's health in the diseased areas.

OBJECTIVETo investigate the correlation between redundant prepuce and premature ejaculation.

METHODSFifty-two cases suffering from premature ejaculation and redundant prepuce were treated with circumcision. They were asked to fill the investigating questionnaire about the changes of ejaculatory latent period, patients' and their wives' satisfaction with sexual life before and after the treatment.

RESULTSDuring 12 months after circumcision, 28 cases were cured and 11 cases were efficacious. The curative rate was 54.9% and effective rate was 76.5%. Twelve cases with no responding continued to be treated with routine methods, such as psychotherapy, daub narcotic to glans of penis and taking medicine to treat chronic prostatitis, which were used before circumcision but still no effects. During 18 months after circumcision, 4 cases were cured and 5 cases were efficacious.

CONCLUSIONRedundant prepuce had direct or indirect relationship with premature ejaculation. The circumcision is one of the effective methods to treat premature ejaculation.

Adult ; Aged ; Circumcision, Male ; Ejaculation ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Sexual Dysfunction, Physiological ; surgery

The effects of human insulin 70/30 and insulin lispro 75/25 were compared in improving postprandial blood glucose excursions in 106 patients with type 1 or 2 diabetes in a one-month,open-labelled,self- controlled trial .The results showed that treatment of diabetic patients with insulin lispro 75/25 significantly improved 2 h postprandial blood glucose excursion compared to pre-study with human insulin 70/30 (baseline) without any significant adverse events or sustained hypoglycemic episodes.These physiological benefits were associated with a patient preference for insulin lispro 75/25.

Purpose: The aim of this study was to compare the efficacy of liver transplantation (LT) and liver resection (LR) for hepatocellular carcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to investigate risk factors affecting prognosis. Materials and Methods: A total of 94 HCC patients with PVTT type I (segmental PVTT) and PVTT type II (lobar PVTT) were involved and divided into LR (n=47) and LT groups (n=47). Recurrence-free survival (RFS) and overall survival (OS) were compared before and after inverse probability of treatment weighting (IPTW). Prognostic factors for RFS and OS were explored. Results: Two treatment groups were well-balanced using IPTW. In the entire cohort, LT provided a better prognosis than LR. Among patients with PVTT type I, RFS was better with LT (p=0.039); OS was not different significantly between LT and LR (p=0.093). In subgroup analysis of PVTT type I patients with α-fetoprotein (AFP) levels >200 ng/mL, LT elicited significantly longer median RFS (18.0 months vs. 2.1 months, p=0.022) and relatively longer median OS time (23.6 months vs. 9.8 months, p=0.065). Among patients with PVTT type II, no significant differences in RFS and OS were found between LT and LR (p=0.115 and 0.335, respectively). Multivariate analyses showed treatment allocation (LR), tumor size (>5 cm), AFP and aspartate aminotransferase (AST) levels to be risk factors of RFS and treatment allocation (LR), AFP and AST as risk factors for OS. Conclusion LT appeared to afford a better prognosis for HCC with PVTT type I than LR, especially in patients with AFP levels >200 ng/mL.

Low-intensity pulsed ultrasound (LIPUS) is a promising therapy that has been increasingly explored in basic research and clinical applications. LIPUS is an appealing therapeutic option as it is a noninvasive treatment that has many advantages, including no risk of infection or tissue damage and no known adverse reactions. LIPUS has been shown to have many benefits including promotion of tissue healing, angiogenesis, and tissue regeneration; inhibition of inflammation and pain relief; and stimulation of cell proliferation and differentiation. The biophysical mechanisms of LIPUS remain unclear and the studies are ongoing. In recent years, more and more research has focused on the relationship between LIPUS and stem/progenitor cells. A comprehensive search of the PubMed and Embase databases to July 2020 was performed. LIPUS has many effects on stem cells. Studies show that LIPUS can stimulate stem cells in vitro; promote stem cell proliferation, differentiation, and migration; maintain stem cell activity; alleviate the problems of insufficient seed cell source, differentiation, and maturation; and circumvent the low efficiency of stem cell transplantation. The mechanisms involved in the effects of LIPUS are not fully understood, but the effects demonstrated in studies thus far have been favorable. Much additional research is needed before LIPUS can progress from basic science research to large-scale clinical dissemination and application.
Cell Proliferation ; Humans ; Signal Transduction ; Stem Cells/radiation effects* ; Ultrasonic Therapy/methods* ; Ultrasonic Waves

Optimal vision and ergonomics are essential factors contributing to the achievement of good results during microsurgery. The three-dimensional (3D) digital image microscope system with a better 3D depth of field can release strain on the surgeon's neck and back, which can improve outcomes in microsurgery. We report a randomized prospective study of vasoepididymostomy and vasovasostomy using a 3D digital image microscope system (3D-DIM) in rats. A total of 16 adult male rats were randomly divided into two groups of 8 each: the standard operating microscope (SOM) group and the 3D-DIM group. The outcomes measured included the operative time, real-time postoperative mechanical patency, and anastomosis leakage. Furthermore, a user-friendly microscope score was designed to evaluate the ergonomic design and equipment characteristics of the microscope. There were no differences in operative time between the two groups. The real-time postoperative mechanical patency rates were 100.0% for both groups. The percentage of vasoepididymostomy anastomosis leakage was 16.7% in the SOM group and 25.0% in the 3D-DIM group; however, no vasovasostomy anastomosis leakage was found in either group. In terms of the ergonomic design, the 3D-DIM group obtained better scores based on the surgeon's feelings; in terms of the equipment characteristics, the 3D-DIM group had lower scores for clarity and higher scores for flexibility and adaptivity. Based on our randomized prospective study in a rat model, we believe that the 3D-DIM can improve surgeon comfort without compromising outcomes in male infertility reconstructive microsurgery, so the 3D-DIM might be widely used in the future.