Objective: To construct the recombinant expressive vector of HSV-2-EGFP for the primary application in rapid,direct and sensitive diagnosis of herpes simplex virus infection. Methods: The gene fragment coding for HSV-2 ICP10 promoter was amplified by PCR,and the PCR product was cloned into pEGFP-1 to construct the recombinant plasmid pICP10-EGFP,which would be identified by DNA sequencing.The recombinant plasmid pICP10-EGFP were transfected into Vero cells by cation lipoid Lipofectamine 2000,then G418 was added to screen the positive cells to obtain stable cell line Vero-ICP10-EGFP.Vero-ICP10-EGFP was infected by various titers of HSV-2,and the EGFP fluorescence was detected under inverted microscope at 6,8 and 10 h post-infection.The specificity of the cell line was detected by infection with human cytomegalovirus(HCMV) and coxsackievirus. Results: The recombinant plasmid pICP10-EGFP was proved to be correct by the double restriction enzyme digestion and DNA sequencing.The Vero-ICP10-EGFP fluorescent emitting cells can be observed as early as 6 h after infection with HSV,with pronounced increase in the intensities at later hours.No induction of detectable fluorescence was detected by infection with HCMV and coxsackievirus 6,10 and 24 h post-infection. Conclusion: This novel GFP reporter system expressing the HSV-inducible EGFP reporter gene might provide a fast,easy and sensitive model for monitoring HSV in clinical specimens.

BACKGROUND:Fracture healing in diabetic patients is usual y unsatisfactory because of hormones and metabolic disorder, and an eventual multiple organ dysfunction resulting from high blood glucose. OBJECTIVE:To dynamical y observe the changes of cytokines during the fracture healing process in diabetic rats before and after insulin treatment. METHODS:A total of 120 Sprague-Dawley rats were included in this study. Of them, 90 rats intravenously injected with 5%tetraoxypyrimidine to induce rat models of diabetes were randomized into insulin treatment and diabetes groups, respectively. The remaining 30 rats were intravenously injected with equal volume of saline and selected as control group. The next day, blood glucose was determined. Healing at 1, 4, and 8 weeks after fracture were observed by the X-ray film. Biomechanical strength of the injured right tibia was measured at 4, 6, and 8 weeks after modeling. Cytokines in the osteotylus were determined by immunohistochemical staining and in situ hybridization technique. RESULTS AND CONCLUSION:The X-ray films showed that the speed of fracture healing in the diabetes group was slower than insulin treatment and control groups. Biomechanical strength of the osteotylus in the diabetes group was significantly decreased compared with the insulin treatment and control groups. However, there were no significant differences in above-mentioned parameters between the control and insulin treatment groups. Bone morphogenetic protein 2, basic fibroblast growth factor, transforming growth factor-beta, and vascular endothelial growth factor were widely expressed in the osteotylus and their expressions in diabetes group were significantly lower and slower than those in the control and insulin treatment groups. There was no statistical difference between control and insulin treatment groups. These results indicate that osteotylus formation speed, biomechanical strength, and growth factor expressions at the fracture site in diabetes rats were decreased compared with normal rats. Insulin treatment can enhance cytokine levels at the fracture site, thereby promoting the osteoblast proliferation and fracture healing.

Objective To assess the efficacy and safety of using a novel channel dilator (the Corsair microcatheter) accompanied with special occlusion guide wires for coronary chronic total occlusion (CTO) recanalization. Methods From 2011 December to 2013 August,we performed 89 cases (the study group) using channel dilator and the new special occlusion guide wires for CTO recanalization. Another 89 CTO lesions treated before using the corsair microcatheter were compared as the control group.We recorded clinical characteristic, outcome of PCI,radiation exposure time, contrast utilization and the procedure time. The MACE rate was monitored during follow up. Results The intracoronary channel dilator accompanied with special occlusion guide wires were inserted into 33 left anterior descending arteries, 17 left circumlfex arteries and 38 right coronary arteries. The success rates of procedure were signiifcantly higher in the study group than in the control group (91% vs. 67.7%, P < 0.05). Procedure and lfuoroscopy time tended to be lower in the study group than in the control group. There were no serious complications related to the catheter and no death case recorded. Conclusions The channel dilator accompanied with special occlusion guide wires may facilitate the conventional approach with a higher success rate in PCI.

The laboratory teaching of bacteriological analysis is very important for training practical analysis professionals.For better quality of laboratory teaching,in this paper,we practice a series of reform facilitating the lab teaching of bacteriological analysis,which is involved in the exploration and study of new curriculum system and teaching evaluation.The educational practices create effective teaching and learning environment and better performance.

Objective To investigate the pathogenesis of small cell lung cancer ( SCLC) and to ex-plore the expression of cell cycle related kinase ( CCRK) in SCLC and its clinical significance.Methods Reverse transcription polymerase chain reaction ( RT-PCR) and Western blot were performed to examine ex-pression of CCRK in SCLC and normal tissues.Results The expressions of gene [(0.51 ±0.11)IU/L] and protein [(0.61 ±0.13)IU/L] of CCRK in SCLC tissues were significantly higher than normal tissues [(0.30 ±0.08)IU/L, (0.34 ±0.09)IU/L] ( P <0.05).The expression of CCRK was closely correlated with the clinical curative effect ( P <0.05 ) rather than the clinical stages ( P >0.05 ) .Conclusions The expressions of gene and protein of CCRK in SCLC tissues were significantly higher than normal tissues. CCRK promoted the occurrence and progress of SCLC.Chem can restrain effectually the excessive expres-sion of CCRK.The expressions of gene and protein of CCRK in the different clinical curative effect group had significant difference.

ObjectiveTo study the apoptosis of CD4 + T lymphocytes and the detection of immune function in patients with pulmonary tuberculosis and explore the clinical significance.MethodsThe mononuclear cells were separated from the blood of the tuberculosis patients or the healthy.The flow cytometry was used to measure the percentage of apoptotic CD4 + T lymphocytes,and the standard of T-lymphocyte subsets were detected by using SAP technology.The red cell immune function were determined by using yeast wreath way.Results The apoptosis rate of CD4 + T lymphocytes and CD8 + T lymphocyte was ( 15.882 ± 4.65 ) ％,and (27.69 ± 0.74) ％.The Immune complex positive rate ( 19.40 ± 0.58) ％ in patients with tuberculosis was significantly higher than those in controls ( P ＜ 0.01 ).C3b receptor positive rate in red blood cells was ( 17.73 ± 0.63 ) ％,( 46.48 ± 1.34 ) ％ in CD3 + T lymphocyte,( 28.12 ±0.69 ) ％ in CD4 + T lymphocyte,and the ratio of CD4/CD8 ( 1.0223 ± 0.09362) in the patients with tuberculosis was lower than the control group( P ＜ 0.01 ).There were certain relationships between the apoptosis rate of CD4 + T lymphocytes and the percentages of CD4 + T lymphocyte,the standard of T lymphocyte subsets and the red cell immune function.ConclusionsThe apoptosis rate of CD4 + T lympho,cytes in patients with tuberculosis were significantly higher than the healthy,which led to reducing the number of CD4 + T lymphocytes.There was positive correlation between red cell immunity and T-lymphocyte immunity,and the immunity in red cell and T- lymphocyte was lower than normal controls,which may be related to the immune pathogenesis of pulmonary tuberculosis.

Refractory Crohn’s-like enterocutaneous fistula indicates the aggressive manifestation and lead to poor prognosis of patients. The development of multidisciplinary strategies for fistula administration largely subjects to the deficiency of animal model for disease remodeling and the underlying pathogenic mechanism. For the purpose, infected anal fistula model was conducted by BLV single-core electrolytic aluminum combined with dextran sodium sulfate. Notably, the inflammatory granulation tissue and inflammatory cell infiltration in the perianal tissue were arised on day 7 of the model by utilizing the Hematoxylin–eosin staining. With the aid of magnetic resonance imaging and signals of high-brightness. We intuitively observed the thickening and edema appeared in the fistula wall, which collectively suggested the formation of a fistula in the perianal area of the rat. Distinguish from the current models of anal fistula modeling including the body surface of fistula, backside of fistula and drainage wire of fistula, our model revealed multifaceted advantages such as quicker generation, higher modeling rate, preferable stability, better consistency, cost-effective, and in particular, more convenient to mimic clinical manifestations of anal fistula.

Background and Objectives: Adipose tissue-derived mesenchymal stem cells (ASCs) are recognized as an advantaged source for the prevention and treatment of diverse diseases including type 2 diabetes mellitus (T2DM). However, alterations in characteristics of ASCs from the aforementioned T2DM patients are still obscure, which also hinder the rigorous and systematic illumination of progression and pathogenesis. Methods: and Results: In this study, we originally isolated peripancreatic adipose tissue-derived mesenchymal stem cells from both human type 2 diabetic and non-diabetic donors (T2DM-ASCs, ND-ASCs) with the parental consent, respectively. We noticed that T2DM-ASCs exhibited indistinguishable immunophenotype, cell vitality, chondrogenic differentiation and stemness as ND-ASCs. Simultaneously, there’s merely alterations in migration and immunoregulatory capacities in T2DM-ASCs. However, differing from ND-ASCs, T2DM-ASCs exhibited deficiency in adipogenic and osteogenic differentiation, and in particular, the delayed cell cycle and different cytokine expression spectrum. Conclusions The conservative alterations of T2DM-ASCs in multifaceted characteristics indicated the possibility of autologous application of ASCs for cell-based T2DM treatment in the future.

Humans ; Coronary Artery Disease/surgery* ; Percutaneous Coronary Intervention ; Heart ; Treatment Outcome ; Coronary Angiography

BACKGROUNDThe purpose of this retrospective study was to compare the surgical outcomes of simple discectomy and instrumented posterior lumbar interbody fusion (iPLIF) in patients with lumbar disc herniation and Modic endplate changes. Our hypothesis was that iPLIF could provide better outcome for patients with refractory lumbar disc herniation and Modic changes (LDH-MC).

METHODSNinety-one patients with single-segment LDH-MC were recruited. All patients experienced low back pain as well as radicular leg pain, and low back pain was more severe than leg pain. Forty-seven patients were treated with discectomy and 44 were treated with iPLIF. The outcomes of both low back pain and radicular leg pain using visual analogue scale (VAS) as well as the clinical outcome related to low back pain using Japanese Orthopaedic Association (JOA) score were assessed before and 18 months after surgery, respectively.

RESULTSBoth low back and leg pain were significantly improved 18 months after simple discectomy and iPLIF. Compared to patients undergoing simple discectomy, low back pain was significantly reduced in patients undergoing iPLIF, but there was no significant difference in leg pain between two groups. Solid fusion was achieved in all patients who underwent iPLIF.

CONCLUSIONSIn patients with LDH-MC, iPLIF can yield significantly superior outcome on the relief of low back pain compared to simple discectomy. Simple discectomy can relieve radicular leg pain as efficient as iPLIF. Accordingly, iPLIF seems to be a reliable treatment for patients with LDH-MC and predominant low back pain.

Adult ; Diskectomy ; standards ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Low Back Pain ; surgery ; Lumbar Vertebrae ; surgery ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; standards