OBJECTIVETo investigate the role of RLIP76 in regulating multi-drug resistance in small cell lung cancer (SCLC), and to analyze the relationship between its expression and prognosis.

METHODSThe expressions of RLIP76 protein and gene were detected by Western blotting and real-time PCR (RT-PCR) in both the chemosensitive SCLC H69 cell line and chemoresistant H69AR cell line, respectively. siRNA was transfected into the H69AR cells to inhibit RLIP76 expression, and eGFP-RLIP76 was transfected into the H69 cells to enhance RLIP76 expression. The drug-sensitivity of cells to chemotherapeutic drugs (ADM, DDP, VP-16) were detected by CCK8 assay. The expression of RLIP76 in the SCLC tissues was detected by immunohistochemistry. The relationship of RLIP76 expression with clinicopathological features and prognosis of the patients was analyzed.

RESULTSThe expression of RLIP76 in H69AR cells was 13.675 ± 0.983, significantly higher than 1.074 ± 0.107 in the H69 cells (P < 0.01). The drug-sensitivities of H69AR cells to chemotherapeutic drugs were significantly increased when the expression of RLIP76 was down-regulated (P< 0.001). The sensitivities of H69 cells to chemotherapeutic drugs ADM, DDP and VP-16 were significantly decreased after transfection with eGFP-RLIP76 up-regulating the RLIP76 expression (P = 0.003). The positive expression rates were 61.3% and 9.4% in the SCLC tumor tissues and para-cancerous tissues, respectively (P < 0.01). The expression of RLIP76 was significantly correlated with clinical stage, chemosensitivity and overall survival of the SCLC patients (P < 0.05).

CONCLUSIONSOur results suggest that RLIP76 is involved in the regulation of small cell lung cancer multidrug resistance. RLIP76 may serve as a potential target gene to evaluate the chemosensitivity and clinical prognostic for small cell lung cancer.

ATP-Binding Cassette Transporters ; metabolism ; physiology ; Antineoplastic Agents ; pharmacology ; Cisplatin ; pharmacology ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Etoposide ; pharmacology ; GTPase-Activating Proteins ; metabolism ; physiology ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Small Cell Lung Carcinoma ; drug therapy ; metabolism ; Transfection ; Up-Regulation

Objective Analyze hepatitis B virus(HBV)infection condition in adults,and provide scientific basis for building immune effects of HB vaccine in adults.Methods In Shaoxing city,randomly select 1309 adults between the age of 18 and 50,draw their venous blood and test HBsAg,anti-HBs and anti-HBc.Results 1236 adults completed the test and the testing rate was 94.42%.The positive rate of HBsAg was 7.61%,9.92% in men and 4.53% in women.Positive rates of HBsAg of different genders and ages showed significant difference(?2=12.49,P

Objective To explore immune effects of HB vaccine and relative strategies of controlling hepatitis B in adults of different ages, genders, and different anti-HBs levels before immunization. Methods In Shaoxing city, 1055 healthy adults aged 18 to 50 with HBsAg negative were randomly selected and inoculated with 10?g/ml indigenous CHO HB vaccine according to the programme of 0,1,6 months. Results 835 adults completed the whole programme, and the total immunization coverage rate was 85.12%. Before and after immunization,the positive rates of anti-HBs were 52.10% and 98.44% respectively, and anti-HBsGMT rose from 14.32mU/ml to 413.98mU/ml. For the two groups, anti-HBsGMT=0 and 0

Objective To establish a standardized middle cerebral artery occlusion (MCAO) model with suture method in rabbits and to investigate the value of the assessment for cerebral ischemia with amplitudeintegrated electroencephalogram (aEEG).Methods A total of 34 male New Zealand white rabbits were randomly assigned to either an MCAO group (n =29) or a sham operation goup (n =5).A model of MCAO was induced with intraluminal suture method and the cerebral function monitor was performed.According to 2,3,5-triphenyltetrazolium chloride staining,the MCAO group was further divided into cortex + basal ganglia infarction,basal ganglia infarction,subarachnoid hemorrhage,and non-lesion subgroups.The differences among the physiological indicators,weight,thread end diameter,and insertion length were compared before and after modeling in all subgroups.Results The success rate of MCAO modeling with suture method in rabbits was 62.07％ (18/29),in which 37.93％ (11/29) involved in the cortex and basal ganglia,24.38％ (7/29) only involved in the basal ganglia,17.24％ (5/29) complicated by subarachnoid hemorrhage,and 20.69％ (6/29) had no infarction.There were no significant differences in the body temperature,heart rate,mean arterial pressure and arterial blood pH,oxygen partial pressure,and CO2 partial pressure among all the subgroups before and after modeling.The weight in the non-lesion subgroup was 2.36 ± 0.10 kg,it was significantly lower than 2.55 ± 0.09 kg in the cortex + basal ganglia infarction subgroup (P =0.001) and 2.50 ± 0.12 kg in the basal ganglia infarction subgroup (P =0.017).The length of suture placement in the cortex+basal ganglia infarction subgroup was 5.59 ± 0.24 cm,and it was significantly less than 6.00 ± 0.50 cm in the subarachnoid hemorrhage subgroup (P =0.036).However,it was significantly longer than 5.20 ± 0.50 cm in the non-lesion subgroup (P =0.033).After modeling there were significant differences in aEEG among all subgroups (F =14.059,P =0.000).Compared to before modeling,aEEG decreased 50.02％ (t =9.573,P ＜ 0.001) and 14.20％ respectively after modeling in the cortex + basal ganglia infarction subgroup and the ganglia infarction subgroup (t =2.908,P =0.027).Conclusions A standardized MCAO model in rabbits may be successfully established with suture method.The significantly decreased aEEG indicates that the MCAO model is successful and the lesions involve in the cortex.

Objective To evaluate the effect of the distal humerus extremity comminuted fracture treated by double plates through oleeranon osteotomy. Method Twenty-eight patients with humerus extremity comminuted fractures were fixed by double plates through olecranon osteotomy followed by early functional exercise. Results All the cases were followed up 9-31 months. The curative effect was assessed by the method of modified Cassebaum rating system. The operation was excellent in 10 cases, good in 12 cases, fair in 5 cases, bad in 1 case. The excellent and good rate was 78.57%(22/28). Conclusion Treatment of humerus extremity comminuted fractures by double plates through olecranon osteotomy followed by early functional exercise are excellent with satisfactory functional recovery.

BACKGROUND: Polyethylene glycol-polyethyleneimine/ferroso-ferric oxide (PEG-PEI/Fe_3O_4) was selected as drug carders in tumor treatment, which can increase drug loading capacity and targeting capacity.OBJECTIVE: To test the toxicity of PEG-PEI/Fe_3O_4 nano-magnetic fluid in vitro and in vivo. METHODS: When the prepared PEG-PEI/Fe_3O_4 nano-magnetic fluid reached nano level, 7702 and HpG2 cell lines were filtrated and diluted in 5-20 multiple, and detected by in vitro MTT toxicity test assay; in vivo hemolysis test and micronucleus test was used to test the toxicity and biocompatibility.RESULTS AND CONCLUSION: MTT assay results indicated that the toxicity grade of PEG-PEI/Fe_3O_4 nano-magnetic fluid to 7702 cell line was 0-1, which was harmless to natural hepatic cells; however, PEG-PEI/Fe304 nano-magnetic fluid had slight bystander restraining effect to HpG_2 cell line. Maximum hemolysis rate of the matedel was 0.372%, which was far less than 5%. The micronucieus test result indicated that PEG-PEI/Fe_3O_4 nano-magnetic fluid had no teratogenicity or mutagenicity.

Objective To study the therapeutic effect of Fe3O4 nanometer magnetic fluid-induced hyperthermia on implanted liver cancer in nude mice under alternating magnetic field. Methods Nude mice model bearing implanted HepG2 was established. Mice were then randomly divided into 3 groups: the blank control group; the magnetic field group; nanometer magnetic fluid group. The magnetic field group were just put under the magnetic field; Nanometer magnetic fluid group received injection of PEG-PEI/Fe3O4 nanometer magnetic fluid under the alternating magnetic field. At the frequency of 40 kHz, and magnetic field of 5 kA/m, 15 minutes one day in the next 14 days. On the 7th day and the 15th day, the changes of tumor volume and weight were recorded, cell apoptosis were observed and recorded and pathological examination was done. Results On the 7th and the 15th day, in the nanometer magnetic fluid group, tumors' volume was smaller and the weight was lighter than other groups, and the tumor inhibitory rate of 54. 20% (t = 14. 506,P ＜0. 01 ) was significantly higher than the control group and the magnetic field group 22. 66% ( t = 7.497, P ＜ 0. 05 ). In the control group, tumor cells grew well, high density, the nucleus engrained, the shape irregular, the nuclear fission clear; compared with the control group, in the magnetic field group, tumor cells scatter thinly, intercellular substance increases, and necrosis area formed;in the nanometer magnetic fluid group, many of tumor cells died, their cell nucleus broke up and vanished,the blood vessel reduced obviously, and the tumor cell spread thinly. Conclusions Under the alternating magnetic field, PEG-PEI/Fe3O4 nanometer magnetic fluid inhibits liver cancer growth in nude mice model of HepG2.

Apoptosis has been considered as the only form of regulated cell death for a long time. However, a novel form of programmed cell death called necroptosis was recently reported. The process of necroptosis is regulated and plays a critical role in the occurrence and development of multiple human diseases. Thus, the study on the molecular mechanism of necroptosis and its effective inhibitors has been an attractive field for researchers. Herein, we introduce the molecular mechanism of necroptosis and focus on the literature about necroptosis drug screening in recent years. In addition, the identification of the critical drug targets of the necroptosis is also discussed.

Objective To optimize the extraction process of the compound herba lysimachiae by orthogonal experiment and response surface method. Methods The sum of the peak areas of multiple representative components in the compound herba lysimachiae was used as the evaluation index. The ultrasonic extraction time, ultrasonic temperature and ethanol ratio were analyzed to optimize the extraction process conditions by orthogonal experiment and Box-Behnken response surface analysis method. Results Ultrasonic extraction time and ethanol ratio are important factors in the extraction process of the compound herba lysimachiae. The effect of ultrasonic temperature in the extraction process is not significant. The optimal extraction condition for compound herba lysimachiaeare is 10 min ultrasonic extraction time at 55 ℃ with 55% ethanol. Conclusion The optimal extraction process parameters obtained by combining orthogonal experiment and Box-Behnken response surface method can be used for the extraction of representative components from the compound herba lysimachiae and pharmaceutical research in the future.

ObjectiveTo establish separate and appraisal methods of murine bone marrow mesenchymal stem cells (MSCs) and optimize the suitable conditions inducting MSCs directional differentiating into adipocytes in vitro.MethodsThe differential adherence to plastic was employed to separate MSCs. CFU-f and successive CFU-f cultures were employed to characterize the potent of proliferation and self-renewal of MSCs. The different adipogenic medium was used as induction for the differentiation of MSCs into adipocytes. The differentiated cells were identified by oil red O immunohistochemistry stain.ResultsThe purified MSCs showed the morphology of fibroblasts. It was found that the number of CFU-f formation depended on the planted number of MSCs. It showed a good relationship. Small type colony of CFU-f had little potent to re-clone, but almost 90% big type colony of CFU-f had the potent to regenerate CFU-f. The MSCs could directionally differentiated into adipocytes induced by different adipogenic medium. But more than 96% MSCs differentiated into mature adipocytes when induced by combined with dexamethasone (DM), 1-methy-3-isobutylxanthine (IBMX), insulin (IS) and indomethacin (ID).ConclusionThe purified MSCs can be harvested by method of differential adherence to plastic, and these MSCs have the potent of proliferation and self-renewal. Moreover, more than 96% MSCs can differentiate into mature adipocytes when induced by combined with DM, IBMX, IS and ID.