Background:Enteroids are considered to be the best tool for studies on intestinal epithelium in vitro and have a widely application prospects,however,there are no associated reports in China. Aims:To establish and optimize the culture technique for enteroids and provide a fantastic platform for the basic research of small intestinal epithelial cells in China. Methods:L-WRN cells were cultured routinely and the conditioned medium with different concentrations of fetal bovine serum(FBS)was collected. Six to eight weeks old C57BL/ 6 mice were sacrificed and 15 cm small intestine from the terminal ileum was removed and cut longitudinally. Crypts were digested with EDTA and then collected and embedded in Matrigel? Matrix;after polymerization of Matrigel? Matrix,L-WRN conditioned medium at different concentration gradient was added. The budding ratio and length of buds were measured dynamically under microscope. The enteroids were re-embedded for subculture when certain length of buds was reached. Results:Compared with L-WRN conditioned medium containing 20% FBS,the conditioned medium containing 10% FBS was more favorable for enteroids culture in vitro. When conditioned medium accounted for 10% ,15% ,20% ,25% or 30% of the mixed medium,they all promoted the growth of enteroids and the 15% one seemed to yield better result. Conclusions:An enteroids culture technique was successfully established for the first time in China. When the L-WRN conditioned medium containing 10% FBS accounts for 15% of the mixed medium,it might promote budding better than the others.

Objective To study the effect of chitosan-pCrmA nanoparticles on the apoptosis of chondrocytes induced by interleukin-1 beta (IL-1β).Methods Chitosan-pDNA nanoparticles were prepared and characterized.The transfection efficiency of chitosan-mediated pIRES2-EGFP was evaluated using fluorescence microscope.The cytotoxicity of chitosan-pIRES2-EGFP nanoparticles in primary rabbit chondrocytes was analyzed by MTT assay.The expression of chitosan-mediated pCrmA in primary rabbit chondrocytes was verified by Western blotting.The effect of chitosan-mediated CrmA on chondrocytes apoptosis induced by IL-1β were analyzed by TUNEL assay.One-way ANOVA was used to analysis.Results The size of chitosan-pDNA nanoparticles was 50 nm.The pDNA release of chitosan-pDNA nanoparticles appeared as biphasic release at pH 2.0 and pH 7.4 buffer.The expression of CrmA in rabbit primary chondrocytes mediated by chitosan could be detected.The chitosan-pIRES2-EGFP nanoparticles had no cytotoxicity.The apoptosis rate of chondrocytes in the chitosan-pCrmA nanoparticles treated group was significantly lower than that of the chitosan treated group (P＜0.05) and PBS group (P＜0.01).Conclsion Chitosan is an effective non-viral gene transfer vector.The CrmA mediated by chitosan can significantly inhibit chondrocytes apoptosis induced by IL-1β,suggesting that chitosan-pCrmA nanoparticles may be the treatment of osteoarthrifis.

Objective To investigate the effect of physician guidance on cigarette cessation in current smokers with chronic obstructive pulmonary disease(COPD).Methods Cigarette cessation education and routine telephone follow-up was provided for 70 COPD patients.Pulmonary function and exhaled carbon monoxide level were detected every 3 months.Results After 6 months' follow-up,28 participants(40.00%) successfully quitted cigarette smoking.Seventeen subjects (24.28%) experienced relapse,with mean cessation time(3.08±0.33) months.Once receiving physician guidance,5 re-attempted to quit cigarette smoking,and 12 reduced tobacco consumption. Sixteen subjects(22.86%) did not quit smoking,although the mean cigarette consumption per day was decreased by 60%.Nine participants (12.86%) showed no change in smoking habit. Conclusion Physician guidance could impreve the rate of cigarette cessation in COPD patients.Intensive intervention may be correlated with successful smoking cessation.

BACKGROUND:Cel cryopreservation is required for clinical use of stem cel s, and the current process of cryopreservation however may be harmful to cel viability, pluripotency and differentiation capacity.
OBJECTIVE:To explore the effect of fructose and dithiothreitol on pluripotency and osteogenesis of cryopreserved bone marrow mesenchymal stem cel s.
METHODS:Bone marrow mesenchymal stem cel s were isolated from the bone marrow of Sprague-Dawley rats and pretreated with fructose (200μmol/L), dithiothreitol (500μmol/L) or combined components before cryopreservation. Then the cel s were cryopreseved for 6 months and the morphology of cel s was observed by inverted microscopy. The cel viability was evaluated by MTT, and real-time PCR was used to detect the mRNA expression of Nanog, OCT4 and Sox2. Alkaline phophatase activity assay and alizarin red staining were utilized to detect the osteogenic capacity of bone marrow mesenchymal stem cel s.
RESULTS AND CONCLUSION:Images captured by inverted microscopy showed no significant difference in cel morphology between groups. The MTT results indicated that fructose and combined pretreatment could promote the cel viability of bone marrow mesenchymal stem cel s after cryopreservation, while the real-time PCR results demonstrated that dithiothreitol significantly facilitated the expression of Naogo and Sox2 in bone marrow mesenchymal stem cel s. Moreover, ALP activity assay and alizarin red staining confirmed the positive effects of fructose, dithiothreitol and combined pretreatment on osteogenic capacity of bone marrow mesenchymal stem cel s after cryopreservation, and the best effects were found after pretreatment with dithiothreitol and combined components. Overal , these findings indicate that fructose pretreatment is beneficial for cel viability of cryopreseved bone marrow mesenchymal stem cel s, and dithiothreitol contributes to maintaining the pluripotency and osteogenesis capacity of cryopreseved bone marrow mesenchymal stem cel s.

Objective To sum up the current status and developmental trend of researches on recuperative medicine worldwide,and to suggest the orientation,aims and focus of research work of recuperative medicine in PLA.Methods Review of published literature was used and on-the-spot investigation was performed to retrieve the publishments in concern and achievements in scientific researches acquired and published in recent years domestically and abroad,and analyze the new progresses and developmental trend of the subject of recuperative medicine.Results With the advances in modern science and technology,recuperative medicine has developed quickly in PLA in recent 5years,and outstanding achievements have been acquired in recuperative management,recuperative rehabilitation,recuperative nursing,recuperative psychology and recuperation for special troops.Conclusion Researches should be done in the future on some core techniques of recuperative medicine,such as natural factor therapy.Meanwhile,more attention should be paid on the recuperative prevention,recuperative health maintenance,recuperative treatment and recuperative rehabilitation for the servicemen who suffered physical and mental injuries induced by high nervous stress and compound risky factors under the condition of high-tech wars on carrying out risky and perilous tasks.

OBJECTIVE To evaluate the clinical outcome of submucosal inferior turbinectomy and outfracture surgery of inferior turbinates. METHODS All patients receiving two different operations were measured by acoustic rhinometry and questionnaire of QOL at preoperative 1 week and postoperative 12 months, seperately. RESULTS Forty-seven patients with inferior turbinate hypertrophy were enrolled in this study. Evaluation of SNOT-20 discovered both surgeries could improve patients' QOL with similar outcome. Preoperative '5 important items' in patients with inferior turbinate hypertrophy were 'lack of a good night's sleep', 'need to blow nose', 'thick nasal discharge', 'fatigue' and 'dizziness'. Postperative '5 important items' were 'postnasal discharge', 'runny nose', 'sneezing', 'reduced concentration' and 'reduced productivity'. Both surgeries could make acoustic rhinometry parameters change obviously, such as minimal cross-sectional area, 0-5 cm nasal volume(NV) and 2-5 cm NV. Furthermore, submucosal inferior turbinectomy produced more volume in nasal cavity than outfacture surgery, (7.28±2.01)cm3 vs (6.01±1.22)cm3, (5.99±1.87)cm3 vs (4.23±1.08)cm3(P<0.05), seperately. There was no correlation between the data of SNOT-20 and acoustic rhinometry. CONCLUSION We recommend outfracture surgery of inferior turbinate as the preferred surgical choice for patients with mild inferior turbinate hypertrophy.

The epitope-G1 gene of Bovine ephemeral fever virus (BEFV) glycoprotein was synthesised by PCR and cloned into expression vector pPIC9K to construct recombinant plasmid pPIC9K-G1. Then the pPIC9K-G1 was linearized and transformed into Pichia pastoris GS 115. The recombinant P. pastoris strains were selected by a G418 transformation screen and confirmed by PCR. After being induced with methanol, an expressed protein with 26 kDa molecular weight was obtained, which was much bigger than the predicted size (15.54 kDa). Deglycosylation analysis indicated the recombinant G1 was glycosylated. Western blot and ELISA tests, as well as rabbit immunization and specificity experiments indicated that the target protein had both higher reaction activity and higher immunocompetence and specificity. The recombinant G1 protein could be used as a coating antigen to develop an ELISA kit for bovine ephemeral fever diagnosis.

Objective To evaluate the effect of CYP3A5 genetic polymorphisms on tacrolimus (Tac) concentration/dosing and other clinical outcomes in a pilot cohort of 113 Chinese HTx recipients.Methods Association between CYP3A5 genetic variants and blood dose-adjusted trough concentrations (C0/D) of Tac at 1st month at the beginning of the immunosuppressive therapy was evaluated in cohorts of 113 patients,then at 1st,3rd,6th,and 12th months after transplantation in 41 patients who received Tac-based immunosuppressive therapy and never changed within one year after transplantation,respectively.In addition,we also evaluated the association between CYP3A5 genetic variants and other clinical outcomes,such as the classifications of endomyocardial biopsy and longterm prognosis.Results The CYP3A5 wild homozygote (* 1/* 1),mutant homozygote (* 3/* 3),and mutant heterozygote (* 1/* 3) occurred in 5,34 and 74 recipients respectively.The gene mutation rate of CYP3A5 in this cohort of Chinese HTx recipients was 80.5 ％ and the homozygous proportion was 65.5％.Compared with CYP3A5 expressors (* 1/* 1 or * 1/* 3),CYP3A5 nonexpressors (* 3/* 3) had a higher Tac C0/D at 1st month (47.34 ± 11.40 vs.116.11 ± 42.40 vs.293.70 ± 171.20,P =0.000),as well as other studied time points (3rd month:98.32 ± 39.43 vs.292.07 ± 141.08,P=0.003;6th month:90.00 ± 21.31 vs.341.68 ± 165.02,P =0.002;and 12th month:96.02 ± 29.33 vs.339.23 ± 162.30,P =0.018);and might have a lower classification of endomyocardial biopsy at 1st month (1.43 ± 0.73 vs.1.50 ± 0.58,P =0.867),3rd month (1.55 ±1.00 vs.2.00 ± 1.73,P =0.512),and 6th month (1.36 ± 0.84 vs.2.33 ± 1.53,P =0.132);as well as a higher mortality due to acute organ rejection (10％ vs.0,P =0.244) and all-cause mortality (20％ vs.9.7％,P =0.580).Conclusion In Chinese HTx recipients,the frequency of this * 3 allele is lower than that has been reported in the white population.The determinations of CYP3A5 genetypes in heart transplant recipients are helpful to guide the individualized Tac regimens.

Objective: To detect the bioavailability of 21 types of perfluorochemicals (PFCs) in rat liver and to examine the effect of protein powder. Methods: Twenty-four rats of the SD strain were randomly divided into the control group, the model group, and the protein powder group. Twenty-one types of PFCs were mixed at an equal concentration of 10 ng/mL, and rats in the model group and the protein powder group were given by oral administration of PFCs mixtures at a daily dose of 5 mL/kg. Rats in the protein powder group were given protein powder by gavage at a dose of 15 mL/kg, while animals in the model and control groups were given deionized water at doses of 15 and 20 mL/kg for 28 successive days. The PFCs contents were quantified in rat liver using ultra-high performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-MS/MS), and the bioavailability was estimated. Results: There were no significant differences in rat body weight or liver/body weight ratio in the control, model and protein powder groups (P>0.05). There were no significant differences in the bioavailability of perfluoroalkylated carboxylic acid (PFCA) or sulfonate (PFSA) in the liver of female and male rats between the protein powder group and the model group (P>0.05), and the gross bioavailability of PFCA (t=-22.266, P<0.001) and PFSA (t=-34.312, P<0.001) was significantly higher in the liver of male rats than in that of female rats in the model group, and the bioavailability of PFCA and PFSA increased followed by a reduction in rat livers with the increase of carbon chain length in the model group. In the model group, the highest bioavailability was measured in perfluorododecanoic acid (PFDoA) and sodium perfluorooctylsulfonate (L-PFOS) in the female rat liver [(36.06±2.93)% and (37.11±1.73)%], and the highest bioavailability was measured in perfluorononanoic acid (PFNA) and L-PFOS in the female rat liver [(61.02±2.16)% and (87.16±3.29)%]. Conclusions The bioavailability of PFCs correlates with the carbon chain length and animal gender in rat livers, and protein powder poses no clear-cut effects on the bioavailability of 21 types of PFCs in rat livers.