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.

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 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 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 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

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.

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.

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.

PurposeThe bone flare phenomena has been well described on bone scintigraphy for efficacy monitoring up to now, but our knowledge has been rarely described on MSCT, the phenomena may be erroneously classiifed as disease progression. This article intends to evaluate the existence and CT features of bone flare phenomena of metastatic bone disease in lung cancer patients treated with ibandronate, to raise awareness of this phenomenon.Materials and Methods The clinical and image data of 45 patients with bone metastases of lung cancer were retrospectively analyzed prior to treatment and 3, 6 months after treatment, the change of CT value and CT features 3 months after treatment between bone flare phenomena group, progressive disease group 1 and progressive disease group 2 were compared.Results The incidence of bone flare phenomena was 6.7% (3/45). 3 months after treatment, CT value of the bone flare phenomena group and progressive disease group 1 changed when compared with before treatment, the differences were statistically signiifcant (t=-5.787 and-2.788,P<0.05) and there was no statistically signiifcant difference (t=1.691,P>0.05) of CT value in the progressive disease group 2 after 3 months' treatment. After 3 months' treatment, the bone flare phenomena group mostly appeared as osteogenic sclerosis of osteolytic lesions, while the cases of progressive disease group mostly appeared as new periosteal reaction of the lesion, or osteogenic/mixed lesion combined with osteolytic damages, the difference between the two groups was statistically signiifcant (χ2=10.139, 8.041 and 4.154,P<0.01,P<0.05). There was no statistically signiifcant difference in increase of density in osteosclerotic lesions (χ2=0.059, P>0.05).Conclusion In patients treated with ibandronate, when there is therapeutic effect evaluation standard of bone metastases (disease progression) and clinical comprehensive curative effect evaluation standard (effective) discordance at 3 months after treatment, it can be interpreted as bone flare phenomena, and the change of CT features contributes to the differential diagnosis of bone flare phenomena with progressive disease.

Objective To develop a microsphere-based suspension array for simultaneous detec-tion and identification of Salmonella H antigens by using Luminex xTAG technology and to evaluate its capa-bility in serotyping Salmonella strains. Methods The fliC and fljB genes, encoding the H antigen of Salmo-nella, were selected as the target genes. Universal upstream primers were designed based on the highly con-served regions of fliC and fljB genes, and the corresponding specific reverse primers were designed based on the variable regions. While synthesizing, the 5′end of each upstream primer was labeled with biotin and the 5′end of each specific reverse primer was modified with its certain TAG sequence. After amplified and la-beled with biotin and TAG sequence, the PCR products of specimens were hybridized with the mixture of va-rious MagPlex-xTAGTM microspheres. Each set of microspheres contained its unique anti-TAG sequences. The results of hybridization were analyzed by using Luminex MagPix reader system and the median fluores-cence intensity ( MFI) was reported. The H antigens of 145 Salmonella strains were identified with this de-veloped xTAG suspension array, and the results were compared with those obtained by using traditional ser-um agglutination test. Results The PCR products of different H antigens ranged from 94 bp to 245 bp and could be identified by hybridizing with MagPlex-xTAGTM microspheres. There was no cross-reaction between different H antigens or with DNAs derived from Escherichia coli, Vibrio cholerae, Vibrio parahaemolyticus and Shigella flexneri. Compared with the traditional serum agglutination test, the sensitivity and specificity of the xTAG suspension array in the identification of H antigens of 145 Salmonella strains were 95. 1% and 100%, respectively. Conclusion The developed xTAG suspension array was a specific, accurate and effective method for simultaneous detection and identification of 31 H antigens of common Salmonella serovars strains. It could be used for determining the H antigens of more than 90 Salmonella strains within 5 hours.