Objective:To investigate the inhibitory effect and its possible molecular mechanisms of MicroRNA-34a(miR-34a) on the human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice.Methods: The human nasopharyngeal carcinoma CNE-2 cell line was cultured in vitro.miR-34a and Scrambled miRNA recombinant plasmids were successfully established and stably transfected into CNE-2 cells.Fifteen six-week-old male nude mice were divided randomly into three groups:miR-34a group(5 mice) ,Scrambled miRNA group(5 mice) ,Blank control group(5 mice).Different CNE-2 cells were subcuta-neously injected on the back near right lower limb.Tumor volumes were examined every 7 days.Mice were executed on the 35 days,and the eventual average tumor volumes and weights were examined.Total RNA and protein were isolated from tumors,and the expression of miR-34a,CDK6,and Bcl-2 mRNA and protein were determined by qRT-PCR and western blot,respectively.Results: The relative expressions of miR-34a was significantly up-regulated in miR-34a transfected group compared to Scrambled miRNA transfected group (P<0.05).Eight subcutaneous xenograft tumor models were successfully established.The nude mice of miR-34a transfected group appeared a smaller tumor volume compared to the other two groups at the beginning of 21th day( P<0.05).The eventual average tumor volumes in miR-34a group,Scrambled miRNA group and blank control group were(351.37±98.19)mm3,(798.75±91.04)mm3 and
(849.62±101.32) mm3 ,respectively,and the eventual average tumor weights in miR-34a group,Scrambled miRNA group and blank control group were(0.81±0.13)g,(1.47±0.21)g and(1.58±0.37)g,respectively.Both the eventual average tumor volumes and weights in miR-34a group were lower compared to the other two groups(P<0.05).qRT-PCR results revealed that the expression of miR-34a in miR-34a transfected group was significantly higher than in the other two groups,while the mRNA and protein expression of CDK6 and Bcl-2 were lower than the other two groups ( P<0.05 ) .Conclusion: miR-34a may inhibit the growth of human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice by down-regulating CDK6 and Bcl-2.

Objective To explore clinical features and misdiagnosis reasons in mitochondrial encephalomyopathy,lactic acidosis,and stroke-like episodes (MELAS) syndrome.Methods The results of clinical data,brain magnetic resonance imaging (MRI),and the course of diagnosis were analyzed in 6 patients with MELAS.Results (1) Clinical features:headache and vomiting were the starting symptoms in 4 of 6 cases,and developmental delay was initial symptoms in 2 of 6 cases.Marasmus occurred in 6 cases,seizure in 5 cases,fever in 3 cases,and hirsutism and visual impairment in 2 cases.(2) Experimental results:blood lactic acid was higher in 6 (4.28 ～ 10.3 mmol/L).(3) Brain MRI:6 patients had abnormal signals in parietal,occipital,temporal lobe,which were not in accordance with vascular distribution.(4) Molecular genetics:All the 6 patients had A3243G gene mutation.(5) Three patients were misdiagnosed for viral encephalitis,and 2 developmental retardation.Conclusions MELAS is characterized with developmental retardation,and repeated encephalitis attack.It is also misdiagnosed because of its variety of clinical features.If patients have high level of lactic acid and multiple MRI signal abnormalities of brain which are not in accordance with vascular distribution,MELAS should be suspected of.Genetic examination and muscle biopsy are especially important in the diagnosis of MELAS.

In the field of colorectal cancer molecular biology,there are a number of biomarkers of prognosis and curative effect,including orotate phosphoribosyltransferase,P53,thymidylate synthase,glutathione S-transferase,methylenetetrahydrofolate reductase,dihydropyfimidine dehydrogenase,uridine-diphosphoglucuronosyl transferase,which can select the effective patients or the patients who can't bear the side effects and contribute to the individualized treatment of colorectal cancer.

Objective To investigate the effect of ramipril on urinary protein in elderly type 2 diabetic patients with nephropathy in different periods.Methods 120 type 2 diabetic patients with nephropathy (64 males, 56 females) with mean aged (68±3) years were randomized into treatment group and control group (n =60, each).According to test results of 24 h proteinuria and renal function, they were divided into 3 subgroups: the normal urine albumin (normal control) group, the early diabetic nephropathy group, and the clinical diabetic nephropathy group.The control group received conventional treatment, while the treatment group used conventional treatment combined with ramipril 2.5 mg/d.Both groups had treatment course of 3 months.The changes in 24 h urinary total protein and urinary albumin before and 1 and 3 months after treatment, and the changes in blood urea nitrogen and serum creatinine levels in patients with renal dysfunction before and 3 months after treatment were observed and compared.Results 24 h urinary total protein and urinary albumin were significantly decreased along with the extended treatment time (P＜0.05 or 0.01).The blood urea nitrogen and serum creatinine levels were significantly declined at 3 months after treatment versus pre-treatment (P＜0.05 for both).There were no significant differences in 24h urinary total protein and urinary albumin in control group before versus after treatment (P＞0.05 for both).At 1 and 3 months after treatment, there were significant differences both in the decrement of 24h urinary total protein and urinary albumin, and in the blood urea nitrogen and serum creatinine levels between the clinical diabetic nephropathy treatment group and the control group (P＜0.05 or 0.01).Conclusions Ramipril combined with conventional treatment can effectively reduce proteinuria and promote the recovery of renal function for type 2 diabetic patients with nephropathy.

BACKGROUND:With the development of stem cellresearch and regenerative medicine, a growing number of technique applications of bone marrow mesenchymal stem cells have been used for treatment of femoral head necrosis. Simple grafting and injection of bone marrow mesenchymal stem cells has a poor effect, which is combined with scaffold materials as cells carrier plays a better role.
OBJECTIVE:To review the research progress of bone marrow mesenchymal stem cells and carriers in the treatment of femoral head necrosis and to look into the future development prospects.
METHODS:A computer-based online search of PubMed (1989/2013) and CNKI (1989/2013) was performed for articles with the key words“osteonecrosis of the femoral head, avascularnecrosis, bone marrow mesenchymal stem cells, carrier or scaffold material”in English and Chinese, respectively. The advantages and limits of various carries and scaffolds were summarized.
RESULTS AND CONCLUSION:Ideal carriers can promote and induce ability of bone marrow mesenchymal stem cells to repair the femoral head. There is a variety of existing carriers, including autologous cancellous bone, al ograft bone, natural biological materials, organic materials, bio-ceramic, nano-materials. Since each material has its own advantages and limitations, composite materials have begun to be used in searching for ideal carriers and scaffolds, and there are however stil some issues that remain to be solved, such as lacking of unified standard of production, specification and making process;how to control and prevent the immune response and inflammation after implantation;how to ensure that the degradation rate of scaffold matches the generation rate of bone and cartilage in the femoral head;and how to identify the cytotoxicity, biocompatibility and therapeutic effects after implantation.

Objective To study the expressions of matrix metalloproteinase-2(MMP-2)and vascular endothelial growth factor (VEGF)in laryngeal carcinoma and their clinicopathological significance.Methods The immuno-histochemical technique was used to detect the expression staining results of MMP-2 and VEGF in 54 cases of laryngeal carcinoma,23 cases of polyps of vocal cord and 1 5 cases of normal laryngeal mucous tissues verified by clinicopathology under the automatic image analyzer.Results The ex-pression rates of MMP-2 and VEGF were 72.22%(39/54)and 70.37%(38/54)in the laryngeal carcinoma tissue,which were sig-nificantly higher than 47.83% and 39.13% in polyps of vocal cord and 40.00% and 46.67% in the normal laryngeal tissues respec-tively,the differences between them had statistical significance(P<0.05).The expressions of MMP-2 and VEGF had certain corre-lation with the clinical stage,lymph node metastasis and.infiltration depth of laryngeal carcinoma.Conclusion MMP-2 and VEGF may jointly participate in the process of infiltration and metastasis in laryngeal and have certain relation with the tumorous invasive growth.

In order to sum up the antineoplasmic active components of Holothuria, we studied the literatures in recent years. The results showed that Holothuria contained a variety of anti-tumor active ingredient, such as Holothurian Glycosaminoglycan and Holothurin. These anti-tumor active components played important roles in different stages of tumor developmment, progression and metastasis. The discovery of new anti-tumor active ingredients may be helpful for the application of Holothuria in clinical tumor treatment.

Objective To evaluate the efficacy of laparoscopic management for bile duct and gastrointestinal injuries during laparoscopic cholecystectomy. Methods Laparoscopic cholecystectomy with or without laparoscopic bile duct exploration was attempted in 9 016 consecutive patients between October 1991 and December 2002. We experienced 14 cases of bile duct injuries (0.15%) and 3 cases of gastrointestinal injuries (0.03%). One patient with circumferential injury and 10 patients with partial laceration injuries of the bile duct underwent a laparoscopic simple closure. One gastric and two duodenal injuries were successfully repaired under laparoscope. Results Expect 1 patient suffered postoperative biliary leakage and bile duct stricture 1 year after the surgery, all the patients were cured and discharged from the hospital. Conclusions Partial laceration injuries of the bile duct or gastrointestinal injuries could be treated with a laparoscopic technique. Concerning laparoscopic management of circumferential bile duct injuries, further study should be taken.

Objective: To analyze the characters of occlusal contact in patients with bruxism.Methods: Occlusal contact was examined by a T-scan II occlusal analyser in 23 cases with bruxism and 13 controls in intercuspal occlusion position, canine protection position, protrusive-retrusive movement and lateral movement. The clusal time, the protrusive disclusion time, the lateral disclusion time and balanced occlusal contacts were calculated and compared between the two groups. Results: The clusion time in bruxism cases was shorter while the lateral disclusion time longer than those in the controls (P

Objective Observing the process of cell aggregation and differentiating into myocardium after the P19 cells treated with different methods and dimethyl sulfoxide(DMSO) inducer.Methods P19 cells were cultivated with DMSO in suspension for 7 days in Petri dishes or dishes containing a thin layer of soft agar to form cell aggregation.Then the aggregates were plated on gelatin-coated culture dishes,and cultured without DMSO to 19 days.The beating of cells was observed.?-sarcomeric actin and cardiac Troponin T(cTnT) immunocytochemical stains were used to identify cell differentiation.Results Cell aggregations were cohering-cultured with growth culture medium after DMSO induced for 7days,spontaneous and rhythmically beating cells were present within the aggregation outgrowths at day 15,which were ?-sarcomeric actin-positive and cTnT-positive.By day 19,positive rates were about 26% and 15% respectively.Conclusion By using DMSO exposure combination prior aggregation can induce P19 cells differentiate into cardiac myocytes which can spontaneously and rhythmically beat.