Objective:To construct an eukaryotic expression vector of enhanced green fluorescence protein(EGFP) gene driven by telomerase catalytic subunit(hTERT) gene promoter and observe the specific expression of EGFP in lung cancer cell lines.Methods:The 1100bp promoter fragment was obtained by enzyme digestion from a recombinant plasmid of pGL3-hTERTp containing the hTERT promoter.The hTERT promoter was then subcloned into the upstream of the report gene EGFP of pEGFP-1 without promoter.The expression vector pEGFP-hTERTp was successfully constructed.The vector pEGFP-N1 containing cytomegalovirus(CMV) promoter was used as a positive control.The vector pEGFP-1 without promoter was used as a negative control.The vectors were transfected into human lung cancer cell lines 95D,NCI-H446,A2,A549,LTEP-a-2,YTMLC and normal MRC-5 through lipofectamine respectively.EGFP expression was detected under the fluorescence microscope.Results:pEGFP-hTERTp was confirmed by enzyme digestion with correct result.That the EGFP expression was detected in all of eight lung cancer cells transfected with pEGFP-hTERTp,but not in MRC cells.By contrast,high intensity EGFP expression was observed in both lung tumor cells and normal cells,which were transfected with pEGFP-N1.Conclusion:The EGFP controlled by hTERT promoter can be expressed specifically in lung cancer cell lines.hTERT promoter may be used as an excellent regulation element in tumor-targeting gene therapy.

Problems in managing laboratory instruments emerges along with the rapid development of laboratory medicine. Full-time instruments manager is positioned in laboratory department, who is in charge of maintaining all instruments. In this way, problems in managing laboratory instruments are solved, so the instruments in laboratory department can be managed in a standard, scientific, institutional and modern way.

Objective:To understand the current situation of hospital violence medical students experienced during the internship period and their professional identity.Method:With the method of cluster sampling,a questionnaire survey was conducted among 500 medical students from a medical college in Ganzhou,to understand the characteristics of hospital violence and compare the difference between the medical students who experienced violence and those not.Results:The effective recovery rate of questionnaire was 90.8％,of which 61.4％ of medical students had witnessed hospital violence,8.1％ of students had experienced hospital violence;the hospital violence that medical students experienced were characterized by low frequency and emotional violence.The overall score of personal identity was (117.05 ± 19.65),was in a medium level.Among the comparison of personal identity between the medical students who had experienced violence and those not,there was a significant difference in occupational cognition dimension (P ＜ 0.01),but there was no significant difference in the total score and other four dimensions (P ＞ 0.05).Conclusion:We should pay attention to the current situation of hospital violence that students experienced and their professional attitude,carry out targeted training and psychological counseling to improve their ability to respond to hospital violence and enhance their professional identity.

Objective To explore the role of aquaporin 1 (AQP1) in the initiation and development of hypertensive disorder complicating pregnancy ( HDCP), and to analyze the relationship between AQP1 expression and ascites formation of patients with eclampsia. Methods Sixty inpatients with HDCP were recruited in the study, including 20 patients with gestational hypertension, 20 with mild preeclampsia, and 20 with severe preeclampsia. And 20 healthy pregnant women were taken as control. Immunohistochemistry was used to analyze AQP1 expressions in placenta, embryolemma and peritoneum, and B-mode uhrasonography was used to detect the ascites level of the patients. Results ( 1 ) AQP1 expression was detected in placenta, embryolemma and peritoneum. AQP1 was mainly located in endotheliocytes of blood vessels and blood capillaries in placenta, endothelial cells of amniotic membrane in embryolemma, and endotheliocytes of blood capillary and small veins in peritoneum- (2)The ascites incidence of HDCP patients ( 63%, 38/60) was higher than that of controls( 10% ,2/20 ; P < 0. O1 ). ( 3 ) The positive expressive rate of AQP1 in placenta of patients with HDCP ( 85% ) was higher than that of controls ( 70%, P < 0. 01 ).Furthermore, the AQP1 positive expressive rate from severe preeclampsia (90%)was obviously higher than that from gestational hypertension patients ( 80%, P <0. 05 ). (4) The AQP1 positive expressive rate in embryolemma from HDCP patients( 87% ) was lower than that of controls ( 95%, P < 0. 05 ). The expressive rate from severe preeclampsia patients (80%) was obviously lower than that from gestational hypertension patients(95%, P <0. 05) and that of controls. (5) The AQP1 expressive rate in peritoneum from HDPC patients(82% )was higher than that of controls(70%, P <0. 01 ). The expressive rate of AQP1 from severe preeclampsia patients ( 90% ) was obviously higher than that from gestational hypertension patients (75%,P <0. 05) and that of controls. Conclusions The expression level of AQP1 of patients with preeclampsia increases in placenta and peritoneum and decreases in embryolemma, and holds correlation with the degree of HDCP. All these suggest that the changes in AQP1 expression may play an important role in the initiation and development of HDCP and may be one of the mechanisms for ascites formation.

Objective To report a case of cutaneous infection due to Trichosporon dermatis. Methods Lesional discharge and tissue were obtained and subjected to microscopic examination, fungal culture and histopathology, respectively. The fungal isolate was then identified with DNA sequence analysis, API 20C AUX system, gelatin liquefaction test, thermal tolerance test. Antifungal susceptibility test was also performed for the fungus. Results A 70-year-old male presented with a 9-month history of ulcerated swelling of the right medial malleolus after plant puncture. Direct microscopic examination of lesional discharge showed no fungal elements, but histopathological biopsy revealed hyphae and spores in the dermis. Yellowish white yeast-like colony grew on Sabouraud dextrose agar (SDA). Slide culture showed pseudohyphae, true hyphae, arthroconidia and blastoconidia. The isolate was identified as Candida humicola by API 20C AUX system, but as T. dermatis by DNA sequence analysis. The strain was unable to liquefy gelatin, could grow at 25 ℃ to 40 ℃, and was sensitive to amphotericin B, itraconazole, voriconazole and nystatin. The skin lesion completely subsided after 4-month treatment with oral itraconazole. Conclusions The isolate is identified as T. dermatis according to morphological features and DNA sequence, which is sensitive to itraconazole.

Objective This study is to identify long-term functional recovery and maturity of regenerated nerve fibers after repairing mouse nerve defects with chitosan/polylactide-co-polyglycolide artificial nerve grafts ( CPANGs ) . Methods Mouse sciatic nerve defects, 2mm in length, were bridged by CPANGs (n=6), with nerve autograft (n=6) and nerve defect (n=6) as controls.Plantar test, electrophysiological examination and laser Doppler perfusion imaging following nerve crush were carried out at 1 year after repair to assess nerve function recovery , while muscle wet weight ratio, histological assessment and transmission electron microscopy were performed to evaluate nerve re -innervation and maturity of regenerated nerve fibers .Results When compared to the autograft group , the CPANG group did not show statistically significant difference in functional recovery in terms of paw withdrawal latency , neurogenic vasodilatation , amplitude and latency of compound muscle action potentials ( CMAPs ) , wet weight ratio of gastrocnemius and tibialis cranialis muscles , number of myelinated nerve fibers and density of unmyelinated axons .However , both these two repair groups exhibited significantly longer CMAPs latency , thinner myelin sheath and a lag-behind shift of diameter distribution of myelinated axons as compared to the normal control .Conclusion At 1 year after the mouse sciatic nerve defect was repaired by CPANGs , sensory and autonomic nerve function , number of regenerated axons and muscle re-innervation degree were recovered to the same extent as nerve autografting , but the regenerated nerve fibers were in a state of immaturity .

Objective To investigate the clinical value of bronchoalveolar lavage fluid (BALF) and serum tumor markers including carcinoembryonic antigen(CEA), cytokeratin 19 fragment (CYFRA21-1) and neuron-specific enolase (NSE) for the diagnosis of lung cancer. Methods The levels of above mentioned tumor markers in the BALF and serum samples of 70 patients with pathologically diagnosed lung cancer and 40 patients with benign lung diseases (BLD) were measured by electrochemiluminescence. Results The levels of 3 tumor markers in the BALF and serum were significantly higher in cancer patients than those with BLD (P

BACKGROUND:Increasing evidence has shown that lovastatin with less toxicity to normal cels has crucial effects on proliferation, apoptosis and differentiation of various cancer cels. However, its roles in glioma stem cels remain unclear.
OBJECTIVE:To explore the effect of lovastatin on proliferation and apoptosis of glioma stem cels.
METHODS: Flow cytometric sorting was used to separate glioma stem cels from human glioblastoma cel line U87. Effects of lovastatin on the proliferation and apoptosis of glioma stem cels were determined by MTT and flow cytometry, respectively. Furthermore, expression levels of Ki67, Bax and Bcl-2 in glioma stem cels treated with lovastatin were detected using western blot analysis.
RESULTS AND CONCLUSION: The CD133-positive glioma stem cels were sorted from human glioblastoma cel line U87 with a positive percentage of 85%. MTT assay showed that lovastatin inhibited the proliferation of glioma stem cels in dose (5, 10, 20 μmol/L)- and time (24, 48, 72, 96 hours)-dependent manners. Flow cytometry analysis showed that 10 μmol/L lovastatin (48 hours) induced apoptosis in glioma stem cels. In addition, the expression level of Ki67 was decreased by lovastatin treatment in a dose-dependent manner, and the Bcl-2 and Bax expression levels were reduced and increased by 10 μmol/L lovastatin treatment, respectively. In conclusion, lovastatin can inhibit cel proliferation and induce apoptosis of glioma stem cels, and lovastatin may be a potential drug for treatment of brain tumors.

Objective To verify the indicator reference value ranges of blood cells analytic report by quoting the WS/T 405-2012 industry standard in Changsha area and to establish the reference value ranges of research parameters suitable for local area.Methods The blood cells analytic results in 3138 cases of reference group were analyzed according to the program of the Formulation of Reference Intervals in Clinical Laboratory Detection Items and the results were compared with the WS/T 405-2012 industry standard.Results WBC,Neu#,Mon%,MCHC,PLT,MPV,PCT,P-LCC,P-LCR and InR‰ in the report parameters and WBC-D,PDW-SD in the research parameters had no statistical differences between males and females(P>0.050);WBC,Neu#,Mon%,MCHC and PLT had no difference between genders,the indicators were narrowed compared with the reference value ranges in standard;the reference value ranges of Neu%,RBC,HGB,MCV and MCH in the male blood cells analytic indicators were narrowed,the lower limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,the upper limits of Bas%,Lym#,Mon# and HCT reference value ranges were elevated,while the upper limits of Eos# and Eos% reference value ranges were decreased,the Lym% lower limit was decreased;Neu%,Lym#,Mon# and RBC in the female blood cells analytic indicators were narrowed,the lower limits of Bas# and Bas% were increased,the lower limits of Eos%,Lym%,HGB,MCV and MCH were decreased,the upper limits of Eos#,Eos%,Lym%,HGB,MCV and MCH were decreased and the HCT upper limit was elevated.Conclusion The Changsha area should have the reference value ranges of selective quoted standards and establishes the reference value ranges suitable for local area or laboratory.

OBJECTIVE To study the radiose-nsitization by Topotecan on human nasopharyngeal carcinoma in nude mice. METHODS ①To study the maximum tolerance dose of TPT and detect the effective rate of TPT and RT on nude mice. ② Plan of radiosensitization practice:53 nude mice xenografts were distributed to 5 groups:RT 20 Gy group,RT 40 Gy group,TPT 12.5 mg/kg group,TPT 12.5 mg/kg+RT 20 Gy group and the controlgroup. After treatment,the volume of tumors were measured every 3 days in order to value the effective rate [complete remission(CR) + partial remission(PR) ]and regrowth delay time(TGD) and to fit the growth curve. RESULTS This study showed that the effective rates had significant difference among RT20 Gy+TPT 12.5 mg/kg group,RT20 Gy group and TPT12.5 mg/kg group,while that of RT20 Gy +TPT 12.5 mg/kg group and RT40 Gy group had no statistical difference. SER reached to 1.34. CONCLUSION Topotecan has been shown a radiosensitizing effect on human nasopharyngeal carcinoma in vivo.