BACKGROUND: It is one of hot topics for the application of neurotrophic factors including neurenergen-3 to promote peripheral neural regeneration nowadays; however, clinical application is restricted to safety and effective administration route. Gene transfection brings a novel thinking and pathway for neurotrophic factors used in clinic.OBJECTIVE: To observe the expression of neural stem cells modified by neurenergen-3 gene after transfection.DESIGN: Completely randomized study.SETTING: Department of Traumatic Orthopaedics and Hand Surgery, the First Affiliated Hospital, Guangxi Medical University;Department of Hand Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Three healthy SD rats of four months old and either gender were selected from Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Recombinant adenoviral expressing vector for transfection of neural stem cells was constructed in Laboratory of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, and the concentration was 0.15 g/L.METHODS: The experiment was carried out in the Orthopaedic Laboratory and Central Laboratory, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from December 2002 to March 2004. Recombinant plasmid pAD-neurenergen-3 containing with green fluorescent protein (GFP) gene was transfectd into primarily cultured neural stem cells by using cationic tiposome interventional method. At 72 hours after transfection, fluorescent inverted microscope was used to observe GFP expression in neural stem cells, and transfection efficiency was measured simultaneously. Expression and transcription of neurenergen-3 gene in neural stem cells were detected at 72 hours, 1 and 5 weeks after transfection by using immunocytochemical stain and reverse transcription polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of neurenergen-3 gene in transfected neural stem cells.RESULTS: ① Neural stem cells transfected by recombinant plasmid pAD-neurenergen-3: GFP expressed on partial neural stem cells at 72 hours after transfection, and the transfection efficiency was 40%. Five weeks later, GFP expression was still observed. ②Transcription and expression of neurenergen-3 gene in neural stem cells: Transcription of neurenergen-3 mRNA was observed in neural stem cells at 72 hours, 1 and 5 weeks after transfection, and expression of neurenergen-3 mRNA was still observed in 5 weeks after trans fection.CONCLUSION: As the carrier of cationic liposome, neurenergen-3 gene can effectively transfect, culture and long-term express neural stem cells via the introduction of adenoviruses.

The photolytic behavior of deoxyadenosylcobalamin and methylcobalamin in water solution was investigated with high performance liquid chromatography. The results indicated that the photolytic cleavage rate increased with the light intensity, according to which a new method was developed to determine the concentration of vitamin B12 in fermentation broth. The samples were completely photolyzed after cell disruption. The content of vitamin B12 was obtained by determining the content of the hydroxycobalamin. The method shows many advantages, such as rapidness, high accuracy and low sample quantity needed, over traditions methods. The developed method may be used in the field of vitamin B12 fermentation.

Objective Under the guidance of CT and three dimensional reconstruction, we made therapeutic plan for lung cancer by radiofrequency ablation (RFA) and observed its effect. Methods From February 2007 to January 2009, we used RITA radiofrequency therapeutic equipment to treat lung cancer under the guidance of 64-slice spiral CT (Siemens) and three dimensional reconstruction of the image. The target temperature is 90℃. Results We performed 29 RFA on 25 patients. In the follow-up, 10 of them showed tumor shrinkage by CT scan. 23 showed lack of tumor-uptake value by SPECT scan and 2 showed lower tumor-uptake value. Conclusion CT-guided percutaneous RFA is safe and practical for lung cancer. It has satisfactory short-term effect to reduce tumor burden.

Objective The aim of the present study was to evaluate the clinical application of 2 μm thulium laser in pulmonary nodules resection under VATS.Methods 61 patients,undergoing thoracoscopic resection of pulmonary nodules in thoracic department of Xuanwu Hospital,were identified between January and December of 2016.Of those,30 underwent 2μm thulium laser dissection and 31 were treated with standard technique by using staplers.In terms of clinical characteristics,including gender,age and smoking history,there is no significant difference between the laser group and the stapler group,but the lesion size was slightly larger in the stapler group compared with the laser group.The lesions of the two groups were almost evenly distributed on the five lobes.Results All the procedures were performed successfully under VATS.The intraoperative evaluation of air leaks demonstrated that less than or equal to 2 grade air leaks were observed in 28 cases in the laser group and in 30 cases in the stapler group.Grade 3 air leaks requiring a rescue treatment were observed in 2 cases in the laser group and in 1 case in the stapler group.There were no significant differences in the postoperative hospital stay and total hospital stay between two groups.Chest tube duration was lower in the laser group compared with the stapler group even if it was not statistically significant (2.71 vs 3.55 days).Hospitalization costs was significantly lower for the laser group.Conclusion The use of 2um thulium laser to prevent intra-and postoperative air leaks and bleeding is effective and makes patients recover quickly,which allows a minimally invasive,accurate and safe application during thoracoscopic resection of pulmonary nodules.

AIM:To study the effect of vitamin K3(VK3) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS:Cell viability was estimated by MTT assay.AO/EB staining was performed to detect apoptotic cells.Apoptosis and the changes of cell cycle were detected by flow cytometry.NAC was used to observe the effect of growth inhibition by VK3.RT-PCR was used to confirm the changes in gene expression.Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA.RESULTS:PC-3M cells growth was significantly inhibited by VK3(≥60 ?mol/L,P

Objective: To monitor the content of bisphenol S ( BPS ) in vegetables and fruits in Henan Province and evaluate the dietary exposure risk of the population, so as to provide the basis for formulating relevant food safety standards. Methods: From 2018 to 2019, 276 samples of vegetables and fruits produced and sold in Henan Province were collected. BPS was determined by isotope dilution ultra performance liquid chromatography triple quadrupole tandem mass spectrometry ( UPLC-MS/MS ) , and the dietary exposure was calculated according to the dietary structure and average body weight of local residents. The risk index of BPS was calculated according to the daily tolerable intake ( TDI ) of bisphenol A ( BPA ). Results: The BPS contents in vegetables and fruits were 0.006-12.600 µg/kg and 0.006-9.380 µg/kg, the medians were 0.053 µg/kg and 0.023 µg/kg, the detection rates were 78.43% and 62.60%, respectively.The detection rate and content of BPS in vegetables were higher than those in fruits ( P<0.05 ). The maximum exposure of BPS from vegetables and fruits was 5.37×10-2 µg/ ( kgbw·d ), and the exposure risk index was 1.07 × 10-3, which was acceptable. Conclusions BPS was detected from vegetables and fruits in Henan Province. The detection rate and content of BPS in vegetables were higher than those in fruits. The health risk of BPS exposed by vegetables and fruits is small.

Background and objective Radiofrequency ablation (RFA) of lung tumours has recently received much attention for the promising results achieved. Here, to evaluate the value of three-dimensional reconstruction CT in radiofrequen-cy ablation (RFA) of advanced non-small cell lung cancer. Methods Sixty-six cases of advanced non-small ceil lung cancer with 68 lesions (2 patients had 2 lesion treated in one session) were underwent three-dimensional reconstruction CT-giuded percuta-neous RFA therapy. To evaluate short-term therapeutic effect of lung tumors using spiral CT scanning in 1-3 months after RFA to investigate the alterations of minor size and density pre-and post-procedure, and complications, to observe the short-term cura-tive effect. Results Our experiences have shown an initial increase in lesion size at immediate follow-up CT. The density of 64 lesions was lowered (94.1%) and 4 lesion is increased (5.9%) at immediate and one month follow-up CT. SPECT scan findings that 82.4% (56/68) cases of FDG uptake in tumors after RFA with tumor/non-tumor of lower than 2.5 at one month follow-up. The change in treated lesion size over time, radiologically assessed through measurements of the lesions on axial CT scans in the lung window setting no lesions had complete response, 50 lesions (73.5%) had partial response, 2 lesions with stable disease, 6 lesions showed progressive disease at 3 month follow-up CT. SPECT scan findings that 79.4% (54/68) cases of FDG uptake in tumors after RFA with tumor/non-tumor of lower than 2.5 at 3 month follow-up. Conclusion The percutaneons RFA therapy under the guidance of three-dimensionsl reconstruction CT scan is safe and effective, with few complications, and can serve as a new method to the treatment of advanced non-small ceil lung cancers.

OBJECTIVETo study the effect of ultrafiltration-membrane extracts of Radix Rehmanniae Praeparata (UMERRP) on theproliferation and genetic stability of bone marrow-derived mesenchymal stem cells (BMSCs) induced by cadmium chloride (CdCl2).

METHODSProtective effects on the proliferation, micronuclear rates, chromosome aberration rates, and apoptosis rates were observed by micronuclei test, karyotype analysis, and flow cytometry.

RESULTSCompared with the CdCl2 group, UMERRP with different molecular weights at 0. 8 g/L could obviously promote the proliferation (P <0. 05). Compared with the control group, micronuclear rates, chromosome aberration rates, and apoptosis rates were obviously enhanced in the CdCl2 group (P <0. 05). Compared with the CdCl2 group, UMERRP with different molecular weights could obviously decreased CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates (P <0. 05). Of them, BMSC micronuclear rates and chromosome aberration rates decreased most obvious in UMERRP groups with molecular weight below 10 000 (P <0. 05). The apoptosis rate decreased most obviously in UMERRP groups with molecular weight ranging 100 000 and 200 000 (P <0. 05).

CONCLUSIONUMERRP could reduce CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates.

Apoptosis ; Bone Marrow ; Cadmium Chloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; Ultrafiltration

BACKGROUNDSlp2 gene is one of the differential expression genes in lung cancer tissues and normal controls, the aim of this study is to observe the effects of antisense Slp2 gene on growth and proliferation in lung cancer cell lines.

METHODSTotal RNAs were extracted from 4 lung cancer cell lines (A549, GLC-82, NCI-H446, NCI-H460), Slp2 gene expressions were detected at mRNA level by reverse transcription-polymerase chain reaction (RT-PCR); antisense Slp2 gene plasmid was constructed and transfected into GLC-82, NCI-H446 and NCI-H460 cells with lipofectinmin. The expressions of Slp2 gene were analyzed by RT-PCR in these cells before and after transfection; Flow cytometry, MTT assay and soft agar culture were used to investigate the effects of antisense Slp2 gene transfection on DNA content changes in cell cycle, proliferation and colony formation ability of these cells.

RESULTSSlp2 gene was positive expressed in A549, GLC-82, NCI-H446 and NCI-H460 cell lines. The proliferation of GLC-82, NCI-H446 and NCI-H460 cells were inhibited by antisense Slp2 gene conspicuously. The flow cytometry experiments showed accumulation of transfected cells in G2-phase, for one time concomitant appearance of apoptotic cells of sub-G1 (hypodiploid) DNA content was observed in transfected NCI-H446 cell. Dramatic inhibition of colony formation in soft agar was observed in those transfected lung cancer cell lines.

CONCLUSIONSThe results suggest that Slp2 gene might be a key gene in lung cancer cell growth and proliferation.

BACKGROUNDPrevious researches about necrotic pancreatic tissue infections are numerous, but the study on systemic infection related to the severe acute pancreatitis (SAP) treatment period is limited. This study aimed to investigate the distribution and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP during the past three years.

METHODSA retrospective study was conducted on the distribution, category and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP from 2008 to 2011.

RESULTSA total of 594 pathogenic bacteria samples were isolated. Among them 418 isolates (70.4%) were Gram bacteria negative, 142 isolates (23.9%) were Gram bacteria positive, and 34 isolates (5.7%) were found fungi. The most common Gram negative bacteria were Escherichia coli (19.8%), and the dominant Gram positive pathogenic bacteria were Enterococcus faecium. The distribution of SAP-related infectious pathogens was mainly in peritoneal drainage fluid, sputum, bile, and wound secretions. Almost all the Gram negative pathogenic bacteria were sensitive to carbapenum. Extended-spectrum β-lactamases (ESBLs) producing strains were more resistant to penicillins and cephalosprins than the ESBLs non-producing strains. Staphylococcus was sensitive to vancomycin and linezolid. The drug resistance of meticillin-resistant staphylococcus (MRS) to commonly used antibiotics was higher than meticillin-sensitive streptococcus (MSS). Enterococcus sp. exhibited lower drug-resistance rates to vancomycin and linezolid.

CONCLUSIONSGram negative bacteria were the dominant SAP-related infection after hepatobiliary surgery. A high number of fungal infections were reported. Drug resistant rates were high. Rational use of antibiotics according to the site of infection, bacterial species and drug sensitivity, correctly executing the course of treatment and enhancing hand washing will contribute to therapy and prevention of SAP-related infection and decrease its mortality.

Anti-Bacterial Agents ; pharmacology ; Gram-Negative Bacteria ; drug effects ; Gram-Positive Bacteria ; drug effects ; pathogenicity ; Humans ; Microbial Sensitivity Tests ; Pancreatitis ; microbiology