Objective To investigate the effects of therapeutic hypercapnia on acute pulmonary allograft rejection induced by macrophages in rats.Methods Twenty-four adult male Wistar rats and 12 adult male Sprague-Dawley rats,weighing 250-280 g,were used in this study.The recipient rats were randomly divided into 3 groups using a random number table (n =6 each):syngraft group (group S),allograft group (group A) and therapeutic hypercapnia group (group H).In group S,Wistar rats served as donors and recipients,while in A and H groups,Sprague-Dawley rats served as donors and Wistar rats served as recipients.Orthotopic left lung transplantation was performed using the cuff technique.After transplantation,the rats inhaled 50％ N2-50％ O2 for 90 min during reperfusion in S and A groups,while in group H the rats inhaled N2-O2-CO2 for 90 min during reperfusion and PaCO2 was maintained at 80-100 mm Hg and O2 concentration in inspired air at 48％-50％ by adjusting the concentrations of the three gases.At 7 days after operation,the arterial blood sample was collected for blood gas analysis and for determination of serum concentrations of tumor necrosis factor α (TNF-α) and interferon γ (IFN-γ)by ELISA.The oxygenation index was calculated.Then the rats were sacrificed,and the transplanted lungs were removed for microscopic examination and for detection of infiltration of macrophages (by immunohistochemistry)and cell apoptosis (by using TUNEL) in lung tissues.The rejection was scored and apoptotic index was calculated.Results Compared with group S,PaCO2,serum concentrations of TNF-α and IFN-γ,rejection score,the number of macrophages and apoptotic index were significantly increased,and oxygenation index was decreased in group A (P ＜ 0.05).Compared with group A,pH value and oxygenation index were significantly increased,and serum concentrations of TNF-α and IFN-γ,rejection score,the number of macrophages and apoptotic index were decreased in group H (P ＜ 0.05).Conclusion Therapeutic hypercapnia can reduce macrophage-induced acute pulmonary allograft rejection possibly through inhibiting the inflammatory responses and cell apoptosis.

Hepatitis B is a serious hazard to human health.This paper describes the current clinical use of several nucleoside antiviral drugs combination,and introduces the candidate drugs which are able to effectively inhibit the hepatitis B virus replication in vivo and in vitro in two years,which provides reference for the research and development of new anti-HBV drugs.

Objective To explore the effect of wrist extensor energy on humeral epicondylitis. Methods 48 humeral epicondylitis patients were divided into muscular energy group and block therapy group with 24 cases in each group. The muscular energy group was treated with muscle energy technique, and the other group received block therapy. They were assessed with Visual Analogue Scale (VAS) and muscle strength. They were followed up 3 months, 6 months and 1 year after discharge. Results The score of VAS was lower in the block therapy group than in the muscular energy group (P<0.01), and the muscle strength was weaker (P<0.01). 3 months after discharge, there was no significant difference in the effects between the muscular energy group (83.3%) and the block therapy group (91.7%) (P>0.05); 6 months after discharge, the muscular energy group (75.0%) was better than the block therapy group (46.1%) (P<0.01); 1 year after discharge,the muscular energy group (54.2%) was better than the block therapy group (16.7%) (P<0.01). Conclusion The block therapy is better in short-term effect on humeral epicondylitis, and the muscle energy technique was better in long-term effect.

Bridged-Ring Compounds ; poisoning ; Child ; Child, Preschool ; Electroencephalography ; Female ; Hemoperfusion ; Humans ; Male ; Prognosis ; Treatment Outcome

Objective We reported previously that single nucleotide polymorphisms SNP) + 11G ＞ A in intron 3 of the human urate transporter 1 (hURAT1) gene are associated with hyperuncaemia in Han Chinese.The aim of the present study was to evaluate the effect of the variants on hURAT1 function.Methods The wild-type,mutant-type hURAT1 and exon 5-null hURAT1 were constructed,and respectively microinjected into the zebrafish embryo yolks.The subcellular localization of different genotypes of hURAT1 was detected by confocal laser scanning microscope.Results Compared with wild type,the mutant recombinant plasmid transcribed two types of mRNA spliceosome,the wild type and the exon 5-null type.The hURAT1 wild type protein was prominent localized on cell membrane,while the mutant type and exon 5-null hURAT1 proteins were distributed uniform in the cytoplasm but not on the cell membrane.Conclusion The hURAT1 variant + 11 G ＞ A resulted in an alternative splicing of hURAT1 mRNA-exon 5-null type.Its protein product exhibited a different subcellular localization compared with that of wild type.

AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.

Objective To observe the location of pelvic organs, the morphology and function of levator ani muscle (LAM) in primiparous women post vaginal delivery at 6 months postpartum using static and dynamic MRI, and investigate the correlation between LAM injury and pelvic organ prolapse (POP). Methods A perspective analysis of static and dynamic MRI was performed in fifty-one primiparous women post vaginal delivery at 6 months postpartum and thirty-five nulliparous women without experience of pregnancy and delivery as control group from June 2014 to January 2015. Previous pregnancy and abortion history, previous pelvic surgery and pelvic mass diseases were excluded. Cases with pelvic floor dysfunction symptoms were excluded from the control group. All of the women underwent static and dynamic MRI. The primiparous group was divided into two groups on presence or absence of POP on MRI findings:primiparous POP group and primiparous control group. The levatorani scoring system based on static MRI was used to characterize morphological changes of LAM into none, minor and major injury by the total score of bilateral LAM. A series of parameters including H line (the distance between the inferior margin of pubic symphysis to anorectal junction), M line (the perpendicular distance between the distal end of H line to pubococcygeal line), levator plate angle (LPA), iliococcygeal angle (ICA), and levator hiatus length and area were measured on static and dynamic MR images. Fisher exact test was performed to compare difference in distribution of the LAM injury between the primiparous group and control group, as well as the primiparous POP group and primiparous control group. Independent sample t test or Mann-Whitney test was used to compare difference in LAM parameters between the primiparous POP group and primiparous control group. Results In the 51 cases primiparous group, 44 cases showed none injury, whilst 5 cases with minor and 2 cases with major injury in the puborectal muscle. Thirty two cases showed none injury, whilst 10 cases with minor and 9 cases with major injury in the iliococcygeal muscle. In the 35 cases control group, none injury was shown in puborectal muscle, whilst 32 cases with none, 2 cases with minor and 1 case with major injury in the iliococcygeal muscle. There was no significant difference in the puborectal muscle injury between the two groups (P=0.203), and there was significant difference in the iliococcygeal muscle injury between the two groups (P<0.05). In the 24 cases primiparous POP group, 20 cases showed none injury, whilst 2 cases with minor and 2 cases with major injury in the puborectal muscle. Fourteen cases showed none injury, whilst 6 cases with minor and 4 cases with major injury in the iliococcygeal muscle. In the 27 cases primiparous control group, 24 cases showed none and 3 cases with minor injury in the puborectal muscle, whilst 18 cases with none, 4 cases with minor and 5 cases with major injury in the iliococcygeal muscle. There was no significant difference in the puborectal muscle injury and iliococcygeal muscle injury between the two groups (P=0.588 and 0.559, respectively). The LH during Valsalva status in primiparous POP group and primiparous control group were (6.7 ± 1.1) and (5.0 ± 0.6) cm, respectively, whilst the LHA was (41.6 ± 12.6) and (24.2 ± 5.5) cm2. There were significant difference between the corresponding groups (P=0.042 and 0.004, respectively). There was no significant difference between the corresponding groups of the other LAM parameters on static and dynamic MRI (all P>0.05). Conclusion Vaginal delivery may cause various degrees of LAM injury, the LAM functional deficiency were observed in primiparous women combined with POP.

Objective To investigate the synthesis, in vivo biodistribution of 99Tcm-HYNIC-PEG4-E[PEG4-c (RGDfk)] 2 (99 Tcm-Galacto-RGD2), and its potential usage for targeted imaging of mice orthotopic glioma.Methods 99Tcm-Galacto-RGD2 was synthesized straightforward and its radiochemical purity and stability and distribution in mice were analyzed.MicroSPECT-CT imaging was done in a mice orthotopic glioma model, which had been set up with U87MG cells, after administration of 99Tcm-Galacto-RGD2.Region of interest (ROI) of glioma was drawn on SPECT-CT section images to quantify tumor uptake (％ ID/cm3).Glioma was harvested for pathological examination.Linear-regression was used to analyze the relationship between integrin αvβ3 and tumor uptake (％ID/cm3).Results The radiochemical purity of 99Tcm-Galacto-RGD2 was (97.7 ±0.8)％ and stable in vitro.Hynic-Galacto-RGD2 could specifically bind to integrin αv β3 of tumor cells with a IC50 of (18 ± 3) nmol/L.After tail vein injection, 99Tcm-Galacto-RGD2 was rapidly discharged from the blood, liver, kidneys and had a relative low concentration in normal brain tissue.MicroSPECT-CT imaging demonstrated that, after 60 min of injection, this drug was well uptaken by glioma tumor than that after 30 min (t =7.13 ,P ＜0.05), and the tumor to normal brain tissue (T/B) uptake ratio of 99Tcm-Galacto-RGD2 was 13.92± 3.43.Injection of HYNICGalacto-RGD2 2 min prior to 99Tcm-Galacto-RGD2 injection extensively reduced the uptake of radioactive drug in tumor tissue (t =11.36, P ＜ 0.05).Bland-Altman analysis showed that tumor volume based on SPECT-CT imaging measurement had almost same value with the tumor reference volume (95％ CI =-11.94％-11.92％).In addition, the tumor uptake of 99Tcm-Galacto-RGD2 and cellular integrin αvβ3 expression level had a linear relationship (R2 =0.896).Conclusions Stable 99Tcm-Galacto-RGD2 can be synthesized easily and is applicable for microSPECT-CT imaging analysis of orthotopic glioma in mice together with the evaluation of integrin αvβ3 level in tumor.

Objective To study the effect of the recombinant Lentivirus containing calcitonin gene related peptide (CGRP) gene on cells biological activity and differentiation of rat bone marrow stem cells(MSCs).Methods Rat MSCs were isolated and cultured by granulocytes adherent.MSCs were transfected with Lenti-EGFP CGRP(MSCsCGRP+/+ group),While MSCs were transfected with Lenti-EGFP as control group.Cell transfection rate was detected by flow cytometry,protein secretion in the above-mentioned MSCsCGRP+ + supernatant was detected using ELISA method.Cells surface markers weare detected by flow cytometry and immunohistochemistry.Trypan blue was used to examin the survive rate,β galactosidase staining was used to examin aging of MSCs transfection,and MTT was used to examine cell vitality.Results At first day after transfecting with Lenti-EGFP-CGRP,fluorescence was not observed by fluorescence microscope,but a small amount of CGRP protein was detected by ELISA in MSCsCGRP+/+ group,at 3 days and 4 days after transfecting with MSCs,strong fluorescence was observed by fluorescence microscope (the cell transfection rates were 77.87％ and 79.58％).The CGRP expression was significantly higher in MSCsCGRP+ + group than in control group [(19.53±0.50) pg/ml vs.(3.12±0.00) pg/ml,t=48.964,P＜0.01].At three days after transfection with MSCs,CD29 and CD90 expression were significantly higher,as compared with control group,CD31 expression was increased in MSCsCGRP+ /+ group.Seven days after transfection with MSCs,CD31 expression was significantly increased in MSCsCGRP+ + group,vWF expression was significantly increased in MSCsCGRP+ + group after MSCs were transfected with LentiEGFP CGRP for 14 days,but a SMA expression was decreased in MSCsCGRP+ +group.At 3 days and 7 days after transfection with Lenti-EGFP-CGRP,the proliferation,survive and aging showed no difference in MSCsCGRP+/+group and in control group (the proliferation of cell:t=0.253,0.290the survive of cell t=-0.307,0.690,all P＞0.05).At 14 days after transfection with Lenti-EGFP-CGRP,aging of cell were decreased in MSCsCGRP+ + group as compared with control group (t=2.446,P＜ 0.05).Conclusions After MSCs are transfected with Lenti EGFP-CGRP,biological characteristics of MSCs has no significant effects,there is still proliferation and differentiation activity.Cell secretion of CGRP can promote the endothelial cell differentiation,and inhibit the differentiation to smooth muscle cells.The CGRP modification of MSCs may play a role in the regulation of angiogenesis.

Objective To test and eva1uate the abi1ity of three potential chloroplast DNA (cpDNA) barcoding sequences;To find new methods to identify the species of gardenia. Methods Three cpDNA sequences were amplified and sequenced by universal primers of matK, rbcL and psbA. By comparing PCR amplification efficiency, length, intra-and inter-specific divergence, and barcoding gap, BLAST and DNA MAN were used to evaluate these loci. Results The amplification efficiency of 5 samples from 3 gardenia species was 100%. Analysis of the intra-and inter-specific divergence of matK among the sequences showed that barcoding gap was superior to psbA and rbcL, with higher identification efficiency. Conclusion Gardenia jasminoides Ellis can be better identified by matK sequence.