AIM: To study the effect of CPUE1 on the reversal of multidrug resistance (MDR) in K562/A02 cells and its mechanism. METHODS: MTT assay was used to determine the influence of CPUE1 on the cytotoxicity of vincrisine (VCR) in K562/A02 cells. The effect of CPUE1 on the apoptosis induced by VCR in K562/A02 cells was detected using DNA analysis and Annexin-Ⅴ/PI double stain assay. Flow cytometry was used to investigate the efflux of rhodamine123 (Rh123) by P-glycoprotein (P-gp) in K562/A02 cells. RESULTS: CPUE1 increased the cytotoxicity and apoptosis induced by VCR in K562/A02 cells. At the concentration of 10 ?mol?L -1 ,CPUE1 reduced the IC_ 50 value of VCR from 60.54 to 4.17 ?mol?L -1 . CPUE1 inhibited the efflux of Rh123 and resulted in the increase of intracellular accumulation of Rh123 in K562/A02 cells. CONCLUSION: CPUE1 has strong reversal effect on MDR in K562/A02 cells by inhibiting P-gp activity.

BACKGROUND: Human adipose tissue-derived stromal cells (ADSCs) possess exert proliferation and multi-directional ability. As a novel stem cell, it has been widely utilized in tissue engineering and plays an important role in biological and potential therapeutic applications.OBJECTIVE: To review the research progress, applications and existing problems of human ADSCs in tissue engineering and cell therapy by retrieving relevant publications. METHODS: PubMed and CNKI databases were undertaken to identify the relevant articles published from January 1960 to January 2009 with the key words of "human adipose tissue-derived mesenchymal cells, isolation, differentiation, immune phenotype, application" both in English and Chinese. The articles relate to biological characteristics and applications of ADSCs were selected. In the same field, the documents published earlier or in the authoritative journals were preferred, and the repetitive studies were excluded. RESULTS AND CONCLUSION: A total of 81 documents were searched by computer, and 57 references were included in the final analysis. The results demonstrated that, human ADSCs share many characteristics, including the high proliferative potential and exhibiting the exert ability to undergo multilineage differentiation under appropriate conditions. Human ADSCs can not only utilize for tissue repairing, but also for cell immune modulation and gene therapy. However, there are still problems in its application. With the development of research on human ADSCs, their biological characteristics will be revealed, and their application in tissue repairing, cell therapy, transplantation, as well as gene therapy must be hold a great promise.

Aim To study the reversal effect of CPUC1 on multidrug resistance in K562/A02 cells. Methods MTT assay was used to detect the cytotoxicity of vincristine. Intracellular accumulation of rhodamine123 was measured by flow cytometry to evaluate the function of P glycoprotein(Pgp). The apoptotic cells induced by vincristine were detected by flow cytometry using DNA content analysis and Annexin V/PI double staining. Results CPUC1 significantly reversed the resistance of K562/A02 cells to vincristine. CPUC1 increased intracellular accumulation of rhodamine123 in K562/A02 cells in a concentration dependent manner. CPUC1 increased the apoptosis induced by vincristine. Conclusion CPUC1 reversed multidrug resistance by inhibiting function of P gp in K562/A02 cells, suggesting that CPUC1 merits further study.

AIM: To investigate the effects of testosterone on endothelial function and intimal proliferation after balloon injury in male rabbit abdominal aorta. METHODS: 24 male New Zealand white rabbits were divided randomly into three groups: control group ( n =8, sham castration), hypotestosteronemia group ( n =8,castration) and testosterone replacement group (n =8,castration +testosterone undecanoate intramuscular injection,14 mg/kg). Abdominal aorta was injured with 3 mm PTCA balloon after testosterone undecanoate had been injected for three days. Two weeks later, blood samples were obtained for detection of plasma testosterone, lipids, metabolic product of nitric oxide (NO - 2/NO - 3), superoxide dismutase(SOD) and malondialdehyde (MDA),and all the abdominal aorta were excised to be analyzed by computer. RESULTS: The intimal area of hypotestosteronemia group were significantly larger than that of other two groups( P

Objective 　Determination of Wilson disease gene mRNA expression in human fibroblast cell strain (Me32aT22/2L) by reverse transcription-polymerase chain reaction (RT-PCR). Methods　Using lipofection reagent, the plasmid vector carrying the Wilson disease gene (pRc/CMV-WD) was transferred into Me32aT22/2L cultured in serum free complement medium. RT-PCR was used to determine WD mRNA expression in Me32aT22/2L. Results　 Wilson disease gene expression was detected in Me32aT22/2L, while no specific signals were detected in untransfected fibroblast. Conclusions　It demonstrated that Me32aT22/2L strain could express the Wilson disease gene, suggesting that Wilson disease gene transfer might develop a new approach to study Wilson disease.

Objective To observe bone formation of compound of autologous periosteum cells plus calcium alginate gelatin in animal body and investigate the best mixing ratio of the two components.Methods A total of 96 healthy adult New Zealand rabbits were randomly divided into six groups (Groups A,B,C,D,E and control group).The excised bilateral periostea from rabbits in each experimental group were all made into 0.4 ml cell suspensions.The compounds were prepared by blending 0.4 ml calcium alginate gelatin with 1,1/4,1/16,1/64 and 1/256 of the periosteum cell suspension respectively,and then were applied to autologous right radial defect area.Gross observation,X-ray examination and histological study were carried out at 2,4,8,12 weeks postoperatively.Levels of serum alkaline phosphatase and serum calcium were determined as well.Results The compounds containing periosteum cell suspension with cell counts of 1/4 or 1/16 and calcium alginate gelatin reached the best osteogenesis.Conclusion Compound of autologous periosteum cell suspension-calcium alginate gelatin induces obvious bone formation and is worthy of clinical application,for it has advantages of satisfactory bone defect repair and easy operation.

Objective To explore the feasibility and superiority of the simple controllable aerocyst pressurization wrapped-up bondage for battlefield first aid in the future high tech local war. Methods The disposable infusion bag is wrapped up in the bottom of the sling in stead of partial surgical dressing. Sphygmomanometer, pressure gauge and heparin hat are connected. Inflate and pressurize the sphygmomanometer and keep the pressure between 3kPa and 5kPa. Results The structure of this bandage is simple and can be operated easily. The pressure is controlled nimblely and conveniently. Except for the accurate effect, it is versatile and inexpensive. In addition, it can be employed in the bone fracture.

Objective To explore the feasibility and superiority of the simplified partial pressurized tourniquet used in bleeding or injured limbs in battlefield.Methods The tourniquet was composed of the disposable infusion bag,bulb for blood pressure apparatus,pressure gauge and heparin cap.The disposable infusion bag wrapped around the bottom of the cravat took the places of the balloon and the harness.The pressure on the upper and lower limbs were 180mmHg and 200mmHg respectively.Results The low-cost tourniquet,with easily adjustable pressure,was easy to operate and carry.It was efficient in hemostasis and had a low rate for complications.Conclusion The tourniquet can be applied to self and buddy aid,and thus the incidence of shock can be decreased.

Objective:To investigate the influencing factors for celiac lymph node metastasis in thoracic esophageal squamous cell carcinoma (TE-SCC), construct a prediction model of celiac lymph node metastasis in TE-SCC, and stratify the probability of celiac lymph node metastasis.Methods:The retrospective case-control study was conducted. The clinicopathological data of 443 patients with TE-SCC who underwent thoracoscopic and laparoscopic esophagectomy with systematic lymph node dissection in the First Affiliated Hospital of Zhengzhou University between March 2015 and April 2019 were collected. There were 259 males and 184 females, aged from 41 to 81 years, with a median age of 64 years. The nomogram prediction model was constructed based on the results of multivariate analysis of influencing factors for celiac lymph node metastasis in TE-SCC, of which calibration curve and decision curve were drawed. The predictive performance was evaluated using the concordance index. The score for celiac lymph node metastasis in TE-SCC predicted by nomogram model was used for further recursive partitioning analysis, and patients were stratified into risk subgroups using the decision-making tree model. Observation indicators: (1) celiac lymph node metastasis in TE-SCC; (2) analysis of influencing factors for celiac lymph node metastasis in TE-SCC; (3) construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC; (4) construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability. Measurement data with skewed distribution were represented as M (range). Count data were represented as absolute numbers and percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data between groups was analyzed using the nonparametric rank sum test. Multivariate analysis was performed using the Logistic regression model. Based on Logistic regression model multivariate analysis, a new nomogram model was constructed using the RStudio 3.4 software. Results:(1) Celiac lymph node metastasis in TE-SCC: celiac lymph node metastasis was found in 89 of the 443 patients, with a celiac lymph node metastasis rate of 20.09%(89/443). (2) Analysis of influencing factors for celiac lymph node metastasis in TE-SCC. Results of univariate analysis showed that tumor location, tumor length, tumor differentiation degree, pathological T staging, nerve invasion, vessel invasion, and thoracic lymph node metastasis were related factors for celiac lymph node metastasis in TE-SCC ( χ2=12.177, Z=-2.754, -4.218, -4.254, χ2=3.908, 33.025, 30.387, P<0.05). Results of multivariate analysis showed that tumor location, vessel invasion, and thoracic lymph node metastasis were independent influencing factors for celiac lymph node metastasis in TE-SCC ( odds ratio=2.165, 3.442, 2.876, 95% confidence interval: 1.380-3.396, 1.787-6.633, 1.631-5.071, P<0.05). (3) Construction of nomogram prediction model of celiac lymph node metastasis in TE-SCC: based on the factors screened by multivariate analysis, including tumor location, vessel invasion, and thoracic lymph node metastasis, the nomogram prediction model of celiac lymph node metastasis in TE-SCC was established, with the concordance index of 0.846. The calibration curve showed a high consistency between the celiac lymph node metastasis probability estimated by the prediction model and the actual rate of celiac lymph node metastasis. The decision curve showed that the nomogram prediction model of celiac lymph node metastasis in TE-SCC had a good prediction value when the probability threshold was 0.001-0.819.(4) Construction of decision-making tree model of celiac lymph node metastasis in TE-SCC and risk subgroup analysis of celiac lymph node metastasis probability: patients were stratified into six risk subgroups using the decision-making tree model based on the celiac lymph node metastasis probability. The group A included patients with no vessel invasion+negative thoracic lymph node, group B included patients with no vessel invasion+the number of positive thoracic lymph nodes of 1-3, group C included patients with no vessel invasion+the number of positive thoracic lymph nodes of ≥4, group D included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+upper or middle thoracic esophageal carcinoma, group E included patients with vessel invasion+the number of positive thoracic lymph nodes of 0-2+lower thoracic esophageal carcinoma, group F included patients with vessel invasion+the number of positive thoracic lymph nodes of ≥3. The group A was low-risk group with the celiac lymph node metastasis probability of 11%, group B and D were intermediate low-risk groups with the celiac lymph node metastasis probability of 27% and 21%, group C and E were the intermediate high-risk groups with the celiac lymph node metastasis probability of 56% and 55%, and group F was high-risk group with the celiac lymph node metastasis probability of 80%. Conclusions:The tumor location, vessel invasion, and thoracic lymph node metastasis are independent influencing factors for celiac lymph node metastasis in TE-SCC. Vessel invasion has the dominant influence on celiac lymph node metastasis, followed by the number of positive thoracic lymph nodes, and then the tumor location. Patients can be stratified into six risk subgroups based on the nomogram prediction model and decision-making tree model of celiac lymph node metastasis in TE-SCC.

Cloning of flanking sequences of double-copy gene is a challenge in molecular biology. We developed a method to solve this problem by combining an optimized inverse PCR (iPCR) with TAIL-PCR. First, Southern blotting analysis was used to determine a proper restriction enzyme that could obtain proper-length restriction fragments that contained the target gene. Then optimized iPCR was performed to amplify the restriction fragments that contained the separated copies of the gene. Based on the obtained sequences, TAIL-PCR was performed to amplify further flanking regions of the gene. With this method, we obtained all of the EcoR I restriction fragments (2.2-5.1 kb) and Hind III restriction fragments (8.5-11.7 kb) of mitochondrial atpA gene in cytoplasmic male sterile (CMS) line and maintainer line of Upland cotton. The results showed that this method was an efficient approach to clone flanking sequences of double-copy gene.
Chromosome Walking ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Genes, Mitochondrial ; Genes, Plant ; genetics ; Gossypium ; genetics ; Plant Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Terminal Repeat Sequences