BackgroundLimbal stem cell deficiency usually leads to blindness, and traditional therapy is limited. Recent research demonstrated that bone mesenchymal stem cells ( BMSCs ) and human amniotic epithelial cells(AECs) could differentiate into many kinds of cells including corneal epithelial cells, but the outcome and effect of these cells on corneal stem cell deficiency are still unclear. ObjectiveThis study aimed to observe and compare the effects of rabbit BMSCs and human AECs transplantation for rabbit limbal stem cell deficiency. Methods Eighteen clean New Zealand rabbits were randomly divided into the amniotic stroma(AS) group, rabbit BMSCs group and human AECs group with 6 rabbits for each group. Limbal stem cell deficiency models were established by putting a piece of filter paper that had been soaked in a NaOH solution at the corneal center. Rabbit BMSCs were isolated and purified by density gradient centrifugation combined with the attachment culture method, and human AECs were collected by a sequential trypsin digestion technique,and the third generation rabbit BMSCs and the first generation human AECs were identified with RT-PCR. After that,cells were inoculated onto the denuded AS and grafted to the corneal surface of the experimental animals. Twenty-eight days after cell transplantation, the therapeutic effects were evaluated based on the corneal neovascularization and opacity scores. Corneal histopathological examination and immunohistochemistry were performed to evaluate and compare the effectiveness among AS,rabbit BMSCs and human AECs on corneal stem cell deficiency. The procedure complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThe third generation of rabbit BMSCs grew well after 12 hours, and the first generation of human AECs formed a membrane-like monolayer after 48 hours of incubation on AS. Immunohistochemistry staining showed that, 28 days after transplantation, the surface cells of the cornea were positive for cytokeratin 3 in both the rabbit BMSCs group and human AECs group.Compared with the AS group,the corneal neovascularization and opacity grades were significantly decreased in the rabbit BMSCs group( Z=-2. 983, P =0. 003 ; Z =-2. 844, P =0. 004 ) and human AECs group ( Z =-2. 817, P =0. 005 ; Z =-2.041, P =0. 041 ). Histopathological analysis exhibited that stratified corneal epithelial-like cells formed on the corneal stroms 28 days after grafting and no signs of goblet cells and neovascularization were found. Less inflammatory cells and regular collagen fiber could be seen in the rabbit BMSCs group and human AECs group. In addition,clinical observation also revealed that the corneas were much clearer in the rabbit BMSCs group than the human A ECs group( Z =-2. 091 , P=0. 037 ), but the corneal neovascularization score was similar between them (Z = -0. 267,P=-0. 789). ConclusionsRabbit BMSCs and human AECs can differentiate into corneal epithelial-like cells onthedamagedcornealsurfaceandfurtherdemonstrateremarkableinhibitoryeffectsoncornealneovascularization and inflammatory cells. The more dominant and prominent effect is the role played by rabbit BMSCsin the improvement of corneal transparency.

OBJECTIVETo investigate the tongue manifestation features of sexually transmitted and intravenous drug use spread HIV infected population in Xinjiang.

METHODSRecruited were 990 HIV infected subjects in Xinjiang from May 2011 to March 2012, who were assigned to the intravenous drug use spread HIV infected (498 cases) and the sexually transmitted (492 cases). By using tongue figure shoot combined with analyses of experts, tongue manifestations were analyzed and compared between the sexually transmitted and the intravenous drug use spread from four aspects, i.e., the tongue color, the tongue shape, the fur color, and the fur property.

RESULTSCompared with the sexually transmitted population, red tongue, fissured tongue, yellow fur, thick fur, eroded fur, deficiency of fur fluid were more often seen, showing statistical difference (P < 0.05). Compared with the intravenous drug use spread population, pale tongue, white fur, and thin fur were more often seen, showing statistical difference (P < 0.05).

CONCLUSIONSThe tongue manifestations of the intravenous drug use spread HIV population reflected inner exuberance of evil toxin and heat impairing qi and yin. Compared with the intravenous drug use spread population, the attack of HIV infection was more hiding in the sexually transmitted population, with milder internal injury. Their Wei-qi was not damaged and no obvious change occurred in the tongue figure.

Adolescent ; Adult ; Aged ; Female ; HIV Infections ; diagnosis ; etiology ; pathology ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Sexually Transmitted Diseases, Viral ; diagnosis ; pathology ; Substance-Related Disorders ; complications ; diagnosis ; Tongue ; pathology ; Young Adult

BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P＜0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P＜0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P ＜ 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P＜0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P＜0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.

Aim To investigate the effects of curcumol on the contractile activity of isolated duodenal smooth muscle in rats and explore its underlying mechanisms. Methods The isolated duodenum specimens of rats were made. The effects of different concentrations of curcumol on the contraction of isolated duodenal smooth muscle were observed using BL-420F biological and functional experimental system with constant tem-perature perfusion method. A certain concentration of curcumol was combined with atropine,isoproterenol, norepinephrine,calcium-free Krebs solution,verapam-il respectively to observe its effect on smooth muscle contraction. Results Curcumol could stimulate duo-denal smooth muscle in vitro,and increase its contrac-tile amplitude and tension significantly. It could be an-tagonized partly by atropine,isoprenaline,norepineph-rine,verapamil. Conclusion Curcumol can promote the contraction of isolated duodenal smooth muscle in rats,which may be achieved by stimulating M recep-tor,inhibiting α and β receptors,and promoting the extracellular calcium influx.

Ribavirin is a broad-spectrum antiviral agent and glycyrrhizin has activities of anti-inflammation, immunoregulation and anti-viral infections. To enhance antiviral efficacy and weaken side-effects of ribavirin, antiviral effects of the combination of glycyrrhizin and ribavirin were studied in the present study. Firstly, a mouse model of viral pneumonia was established by inoculation of influenza H1N1 virus. Protective effects of glycyrrhizin and ribavirin used alone or in combination against H1N1 virus infection in mice were evaluated based on the survival rate, lung index and virus titer in lungs of mice. Results showed that the combination of glycyrrhizin and ribavirin significantly inhibited the lung consolidation with a 36% inhibition ratio on the lung swell of infected mice. The combination of the two drugs exhibited synergetic effects on survival of infected mice. The combination of 50 mg · kg(-1) · d(-1) glycyrrhizin and 40 mg · kg(-1) · d(-1) ribavirin resulted a 100% protection for infected mice with a synergetic value of 36, which was significantly higher than the control group and each drug alone. This combination also resulted a significant drop of lung virus titer (P < 0.01), as well as inhibition on the production of proinflammatory cytokines IL-6 (P < 0.01), TNF-α (P < 0.01) and IL-1β (P < 0.05) induced by virus infection compared to the control. The treatment of ribavirin plus glycyrrhizin was more effective in influenza A infection in mice than either compound used alone, which suggested a potential clinical value of the combination of the two agents.
Animals ; Antiviral Agents ; pharmacology ; Disease Models, Animal ; Drug Synergism ; Drug Therapy, Combination ; Glycyrrhizic Acid ; pharmacology ; Inflammation ; immunology ; Influenza A Virus, H1N1 Subtype ; drug effects ; Interleukin-1beta ; immunology ; Interleukin-6 ; immunology ; Lung ; immunology ; virology ; Mice ; Orthomyxoviridae Infections ; drug therapy ; Pneumonia, Viral ; drug therapy ; Ribavirin ; pharmacology ; Tumor Necrosis Factor-alpha ; immunology

Objective: To observe any curative effect on dysphagia of freehand respiratory training applied to hemiplegic stroke survivors. Methods: Forty-eight hospitalized, hemiplegic stroke survivors were randomly divided into an observation group and a control group, each of 24. Both groups received routine stroke rehabilitation, including motor function training, exercise training, physical therapy and swallowing training. The observation group was additionally provided with barehanded breathing function training, mainly consisting of cough glottis exercise, lip constriction aspiration exercise, chest movement exercise, abdominal muscle contraction and relaxation exercise, as well as abdominal aspiration training. The intervention lasted for 6 weeks for both groups. Before and after the intervention the maximum breathing time, first and second forced volume (FEV₁), forced vital capacity (FVC), maximum inspiratory pressure (PImax), standardized swallowing assessment (SSA), Kubota drinking water test (KDWT) and functional dysphagia scale (FDS) scoring were quantified for both groups. Results: There was no significant difference between the two groups′ averages on any of the measurements before the intervention. Afterward, however, the average maximal breathing time, FEV₁, FVC, and PImax of the observation group were all significantly higher than before the intervention and significantly higher than those of the control group. After treatment, the observation group′s average KDWT efficiency was not, however, significantly better than that of the control group. Moreover, after the treatment the average SSA and FDS of the observation group were significantly lower than those of the control group, as well as significantly better than before the treatment. Conclusion Barehanded breathing function training can supplement routine rehabilitation to significantly improve the breathing of stroke survivors.

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans

OBJECTIVETo explore seroepidemiological status and vaccine coverage of hepatitis B in children aging under 15 years old in Chaoyang district of Beijing.

METHODSA total of 1602 children aging under 15 years old, residents or floating population who had lived here more than six months, were randomly selected by multistage cluster sampling, from Chaoyang district of Beijing in year 2010. The demographic information and vaccine coverage of hepatitis B vaccine (HepB) were collected by self-designed questionnaire.5 ml blood was collected from each subject and the serum HBsAg, anti-HBs and anti-HBc were detected by Abbott microparticle enzyme-linked immunoassay. Those whose HBsAg was positive were then tested HBeAg and anti-HBe. The positive rate of hepatitis B indicators and coverage rate of HepB in different population were compared.

RESULTSThe positive rate of HBsAg, anti-HBs and anti-HBc were 0.56% (9/1602), 64.17% (1028/1602) and 2.12% (34/1602), respectively; while the age standardized rates were separately 0.57%, 66.36% and 1.98%; and the gender-adjusted rates were 0.56%, 64.23% and 2.12% respectively. The positive rate of anti-HBs was statistically significant (χ(2) = 165.445, P = 0.000). The positive rate of anti-HBs was up to 90.73% (235/259) among 1-2 years old children, followed by 76.22% (141/185) among 13 - 15 years old children, 67.21% (166/247) among 3 - 4 years old children, 61.22% (150/245) among 9 - 10 years old children, 60.68% (142/234) among 11 - 12 years old children, 49.05% (103/210) among 5 - 6 years old children and 40.99% (91/222) among 7 - 8 years old children. The average coverage rate of HepB was 90.44% (1371/1516), separately 93.76% (661/705) in residents and 87.55% (719/811) in floating population. The difference was statistically significant (χ(2) = 16.829, P = 0.000).

CONCLUSIONHBsAg positive rate in children under 15 years old in Chaoyang district of Beijing dropped to less than 1% and the coverage rate of HepB had reached over 90%. It is suggested that we should pay more attention to increase the coverage rate of HepB among floating children under 15 years old.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Hepatitis B ; epidemiology ; prevention & control ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; administration & dosage ; Humans ; Infant ; Male ; Seroepidemiologic Studies ; Vaccination ; statistics & numerical data

OBJECTIVETo explore the family aggregation and risk factors of hepatitis B virus (HBV) transmission in Chaoyang district of Beijing.

METHODSA total of 5266 families were randomly selected for the multi-stage cluster sampling study in Chaoyang district of Beijing in 2010. The family members who aged between 1 and 70 years old and lived constantly in Beijing for over half a year, were recruited as subjects. There were 14 491 subjects in total, including temporary residents who did not have Beijing household account, except foreigners. 5 ml venous blood was drawn from every subject. A self-designed questionnaire was used to collect the basic information of the population and the risk factors of the hepatitis B transmission. Microparticle enzyme-linked immunoassay was applied to test five indicators of hepatitis B. Negative binomial distribution test was used among the HBsAg positive families to calculate the family aggregation rate of hepatitis B. Single factor analysis and multi-factor logistic regression model were used to analyze the risk factors of HBV transmission.

RESULTSIn all, 308 out of 5266 families had HBsAg positive members, accounting for 5.85%.383 out of 14 410 subjects were HBsAg positive, rating at 2.66%. The HBsAg positive rate among subjects under 14 years old was the lowest, at 0.56% (9/1603); and the positive rate among subjects aging between 35 and 44 years old was the highest, at 4.27% (47/1029). Negative binomial distribution test showed that the family aggregation rate of HBV infection was 7.66% (χ² = 15.10, P < 0.05). The analysis of family aggregation of HBsAg positive showed that 17.39% (8/46) of the transmission was from father to child, 13.04% (6/46) was from mother to child, 30.44% (14/46) was between couples, and another 39.13% (18/46) was between siblings or other relatives. Both single factor analysis and multi-factor logistic regression analysis showed that hepatitis B positive family members (OR = 5.40, 95%CI: 5.24 - 5.55), hepatitis B positive friends and colleagues (OR = 1.55, 95%CI: 1.11 - 1.99) and blood donation and transfusion history (OR = 1.96, 95%CI: 1.76 - 2.15) were the risk factors of HBV infection.

CONCLUSIONHBV transmission showed family aggregation in Beijing, however, the risk factors needed further studies.

Adolescent ; Adult ; Aged ; Carrier State ; Child ; Child, Preschool ; China ; epidemiology ; Family Characteristics ; Female ; Hepatitis B ; epidemiology ; transmission ; Hepatitis B virus ; Humans ; Infant ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo investigate the characteristics of circulating type II pre-dendritic cells (pDC2) and evaluate its role in patients with chronic hepatitis B virus infection.

METHODSThe quantitative alterations of pDC2 in 27 chronic HBV-infected patients as treated group and 15 healthy individuals as a control group were analyzed by using flow cytometry based on the comparison of CD4+/CD8+ ratios of T lymphocyte subsets between the two groups. The IFN-alpha-producing ability of pDC2 after incubation was determined by ELISA.

RESULTSThe percentage of pDC2 (0.096 +/- 0.086) from the peripheral blood in chronic HBV-infected patients were significantly lower than that (0.304 +/- 0.093) from the normal controls (P less than 0.001) while the CD4+/CD8+ ratios were higher than those in normal controls (P less than 0.01). The values of IFN-alpha-producing function and IL-12 of circulating pDC2 in chronic HBV-infected patients group were significantly lower than those in healthy subjects (P < 0.001). The percentage of pDC2 and CD4+/CD8+ ratios were higher in the patients positive for HBV DNA in sera than those in patients negative for HBV DNA in sera (P < 0.01).

CONCLUSIONThe decreased number of circulating pDC2 and IFN-alpha-producing function from peripheral blood in patients with chronic hepatitis B virus infection may result in the decline of host immune response, which may partially contribute to the disease progress of HBV infection and existence of viral genomic DNA in patient's sera.

Adolescent ; Adult ; CD4-CD8 Ratio ; Cell Count ; DNA, Viral ; blood ; genetics ; Dendritic Cells ; cytology ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Hepatitis B virus ; genetics ; growth & development ; Hepatitis B, Chronic ; blood ; immunology ; virology ; Humans ; Interleukin-12 ; biosynthesis ; Male ; Middle Aged ; T-Lymphocyte Subsets ; cytology ; immunology ; Young Adult