Objective To estabolish an optimal animal model of oxygen-induced retinopathy(OIR) suitable for examining pathogenesis and therapeutic intervention for retinopathy of prematurity(ROP). Methods Fifty-four 7-day-old C57BL/6J mice were devided into two groups. Twenty-seven mice in hyperoxic group were exposed to 75% oxygen for 5 days and then to room air for another 5 days. Twenty-seven mice in normoxic control were exposed to room air for 10 days. The proliferative neovascular response was estimated by observing the vascular pattern in adenosine diphosphate-ase(ADPase) stained retina flat-mounts and quantitated by counting the number of new vascular cell nuclei extending into the internal limiting membrane in cross-sections. Results Angiography in ADP ase stained retina flat-mounts delineated the entire vascular pattern. Hyperoxia-induced neovascularization occurred at the junction between the vascularized and avascular retina in the mid-periphery in all mice exposed to hyperoxia. After 5 days of exposure to hyperoxia at postnatal day 12(P12),the larger central radial vessels became tortuous and constricted and central perfusion became decreased obviously. After return to room air for 2 days at P14,neovascularization was seen. This response was maximal at P17. There was a mean of 44 neovascular nuclei per cross-section extending into the vitreous in hyperoxia compared to less than 2 nuclei in the normoxia control ( P

Objective To investigate the effects of spleen preservation on hepatic fibrosis and relevant cytokine in rabbits with advanced schistosomiasis. Methods After hepatic cirrhosis was induced by infecting Schistosoma japonicum cercariae in rabbits, total splenectomy (TSG), subtotal splenectomy (SSG) or sham operation (model control group, MCG) were performed respectively on these rabbits. Meanwhile,a normal control group (NCG) was established. The serum levels of tumor necrosis factor alpha (TNF-α) , interleukin-6 (IL-6) and interleukin-1 beta (IL-lβ) were detected respectively by radioimmunoassay(RIA) at the 8th, 15th and 21st week post-infection. The expressions of transforming growth factor betal (TGF-β1), type Ⅰ and type Ⅲ collagen in liver tissues were determined by immunohistochemistry before and after the operations. Results Compared with NCG, the serum levels of TNF-α, IL-6 and IL-1β of MCG rabbits increased significantly at the 8th week post-infection (P <0.01). However, the levels of them decreased to a lower level at the 15th week. At the 6th week after operation,no significant difference was found among the three model groups ( MCG, TSG, SSG) (P > 0.05). The expressions of TGF-β1, type Ⅰ and type Ⅲ collagen in liver tissue of MCG rabbits were significantly higher than those of NCG rabbits before the operation (P < 0. 01). No significant difference was found among the three model groups at the 6th week after the operation ( P > 0.05). Conclusion The residual splenic tissue after subtotal splenectomy does not aggravate the hepatic fibrosis at advanced schistosomiasis. The mechanism may be that the relevant cytokines of hepatic fibrosis (TGF-β1, TNF-a, IL-6, IL-1β) decreased to a lower level at this time,and splenectomy does not influence the levels of them.

ObjectiveTo compare the therapeutic efficacy between warm needling and Western medication in treating chronic atrophic gastritis.MethodSixty patients were randomized into an observation group and a control group, 30 cases in each group. The observation group was intervened by warm needling, selecting Guanyuan (CV4), Qihai (CV6), Zusanli (ST36), Xuehai (SP10), and Geshu (BL17); The control group was by Western medication, including Omeprazole, Amoxillin, Metronidazole, etc. The clinical efficacies were compared after 8-week treatment. Besides, gastroscopewas ordered at outset and after 8-week treatment to compare the therapeutic efficacies.ResultIn comparing the clinical symptoms, the total effective rate was 93.3% in the observation group versus 70.0% in the control group (P<0.05); the gastroscope scores dropped significantly in both groups after 8-week treatment (P<0.05), and the score of the observation group was significantly lower than that of the control group (P<0.05). ConclusionAcupuncture-moxibustion can significantly improve the clinical symptoms and gastroscopic results in patients with chronic gastritis, and its therapeutic efficacy is superior to that of Western medication.

Objective To investigate the drug resistance of 1 031 Mycobacterium tuberculosis to rifampicin and isoniazide in the Center Hospital of Changsha from January 1 ,2013 to September 30 ,2014 .Methods A total of 1 031 strains with positive culture result and identified as strains of Mycobacterium tuberculosis were used absolute concentration method to do the conventional drug susceptibility ,and detected rifampicin and isoniazide resistance gene including rpoB ,katG and inhA gene locus mutation by chip technology ,the results of two methods were compared using card square test statistics .Results By gene chip method ,the sensitive strain of rifampicin was 896 ,the drug-resistant strains was 135 ,the sensitive strains of isoniazide was 901 strains ,130 drug-resistant strains .Compared with the absolute concentration method ,resistance chip detection results were consistent with rifampicin resistant strains 1 011 strains(including 894 drug-resistant strains ,and 117 sensitive strains) ,the coincidence rate was 98 .00% ,consistent with isoniazideresistant strains 1 005 strains(including 890 drug-resistant strains ,115 sensitive strains) ,the coincidence rate was 97 .48% .The most common spot of rifampin resistance related mutations of rpoB gene was 531TCG to TTG ,accounted for 51 .11% ,followed by 526CAC→TAC ,accounted for 10 .37% ,11 strains with 526TCG to TTG ,accounted for 8 .15% .Isoniazid re-sistance was caused by mutations in katG315AGC→ACC resistant strains ,accounted for 83 .85% ,inhA-15C→T mutations accoun-ted for only 12 .30% .Conclusion The results of gene chip method is highly consistent with that of absolute concentration method , could be a fast and effective method for screening rifampicin and isoniazide ,the resistant gene of Mycobacterium tuberculosis to rif-ampicin and isoniazide almost mutate in rpoB531 ,526 and katG315 in Changsha .

Objective To study the correlation between the rescue time and the spot survival rate.Method The data of spot-rescued victims in a large public place of Dujiangyan City from 14:35 on May 12,2008 to 11:40 on May 15,2008 were analyzed.The searched-out victims included the spot death and spot survival,and they were statistically analyzed with Chi-Square test and Partitions of X2 method in order to find out correlation between rescue time and survival rate.Results Out of the 366 spot-rescued victims from the ruins,87 ones survived and the spot survival rate was 23.77%.The spot survival rate in the first 24 hours was much higher than that in the second 24 hours(X2=22.62,P＜0.0125)and that in the third 24 hours(X2=37.84,P＜0.0125),and no obvious difference in the spot survival rate between the second and the third 24 hours was found(X2=1.92,P＞0.0125).The first 24 hours was further divided into 3 periods in equal length of time in order to find more subtle differences in early rescue.The spot survival rates in the first and the sccond 8 hours were much higher than that in the third 8 hours(x2=19.33 and 7.11,respectively,P＜0.012 5)while there was no statistical difference in the spot survival rate between the first 8 hours and the second 8 hours(X2=1.75,P＞0.012 5).Conclusions The"golden time"for spot rescuing the victims is the first 24 hours after seismic disaster,the chances to find the survivals is decreasing as the time elapsing.The earlier spot rescue starts in the first golden24 hours,the higher spot survival rate of the seismic victims will be.

Objective This study was aimed to characterize the swine leukocyte antigen( SLA) class I genes of GGTA1 -/ - Wuzhishan minipigs and compare their similarity to human leukocyte antigen( HLA) . It has important implica?tions for understanding the cellular rejection in xenotransplantation. Methods Specimens of ear tissue from six founding GGTA1 -/ - Wuzhishan minipigs were collected, and the SLA class I genes (SLA?1, SLA?3, SLA?2) were amplified by RT?PCR. Purified products were cloned into pEASY?T1 vectors and sequenced, followed by BLAST alignment and using bioin? formatc analysis to characterize the SLA class I genes and compare with the similarity to HLA. Results A total of six al?leles were detected, among them alleles were previously reported (SLA?1?0703,SLA?2?1102, SLA?3?0401, SLA?3?0403), and the other were novel (SLA?1?0401wz01, SLA?2?11wz01). The homology between alleles of SLA class I genes in Wuzhishan minipigs and HLA was from 70?5% to 72?1%. The homology analysis of critical amino acid residues on HLA binding with human CD8 + molecules showed that SLA?1?0401wz01, SLA?1?0703, SLA?2?11wz01, SLA?2?1102 and SLA?3?0401 occurred mutant at amino acid positions 225 and 228 ( T→S,T→M) , whereas the other loci were highly conserved. There was a high homology at amino acid level between SLA?2?11wz01, SLA?2?1102 and HLA class I genes which are NK cell KIRs binding sites. Conclusions The amino acid sequences of SLA class I genes of GGTA1 -/ -Wuzhishan minipigs have a high homology to HLA. From the point of view of cell?mediated xenograft rejection, the amino acid sequences of SLA class I genes of GGTA1 -/ - Wuzhishan minipigs have a high homology to HLA, therefore, Wzhishan minipigs may become a good potential donor for pig?human xenotransplantation.

Objective To summarize the treatment experience of refractory systemic juvenile idiopathic arthritis (JIA) by tocilizumab, and to explore the cost-effective treatment. Methods The clinical data of 6 pediatric patients with refractory systemic JIA treated by tocilizumab from 2014 to June 2016 were retrospectively analyzed in the aspects of course and effectiveness of tocilizumab, steroid reduction, adverse reaction, and growth. Results The median age of the six patients (3 males and 3 females) was 6 years, and the course of disease were from 16 to 63 months. All patients were treated by other immunosuppressive agents or biological agents in addition to steroid and traditional anti-rheumatic drug therapy. The courses of tocilizumab treatment were from 7 to 26 months and the median time was 9.5 months. All 6 patients responded to tocilizumab and achieved the clinical remission at different time. After the induced remission, the interval of the treatment intervention was increased from 2 weeks up to 4 weeks in 3 cases, and no disease activity was observed. Except one case, another 5 cases reduced and stopped the use of hormones at 5.8 months after tocilizumab treatment. After hormones was reduced and discontinued, the growth was improved. All 6 patients had no serious adverse reactions. Conclusions Tocilizumab is safe and effective for patients with refractory JIAs. The steroid can be reduced in short time to improve growth. After remission is induced, the interval of the treatment intervention could be prolonged.

Objective To investigate the effect of artesunate (ASN) on the expression of Heme oxygenase-1 (HO-1) in THP-1 cells induced by the early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) antigens of Mycobacterium tuberculosis and to investigate its possible mechanism.Methods THP-1 ceils were cultured in vitro.The effects of ESAT-6 and CFP-10 on cell viability were detected by methyl thiazolyl tetrazolium (MTT) assay.THP-1 cells were pre-treated with or without ASN prior to incubation with or without ESAT-6 and CFP-10,the mRNA expression of HO-1 was detected by real time quantitative polymerase chain reaction (RT-qPCR) and Toll-like receptor 2 (TLR2) level was measured by Western blot.Results MTF assay showed that ESAT-6 and CFP-10 were non-toxic to cells in the range of 0-5 μg/ml.Compared with the control group,5 μg/ml ESAT-6 and 5 μg/ml CFP-10 could significantly increased the mRNA expression of HO-1 (P ＜ 0.05).In addition,20 μg/ml ASN could significantly enhance the mRNA expression of HO-1 induced by ESAT-6 and CFP-10,and inhibit the expression of TLR2 induced by ESAT-6.Conclusions ASN in combination with ESAT-6 or CFP-10,may have potential value in treatment of pathogen-associated inflammatory diseases.

Objective To explore the laboratory culture and identification of Mycobacterium tuberculosis L forms (MTB-L),isolation rate and drug resistance in smear-positive tuberculosis patients,and to improve clinical attention to MTB-L.Methods 222 smear-positive pulmonary tuberculosis patients treated in our hospital from September 2017 to December 2017 were randomly selected for MGIT 960 and 92-3TB-L liquid culture.After MGIT 960 was reported positive,acid-fast staining was performed on the precipitated smears of 92-3TB-L liquid medium for preliminary screening.The suspected L-positive strain culture was transformed into improved TSA-L solid medium to observe the colony characteristics and microscopic characteristics.The properties of the strain were confirmed by acid-fast staining and tuberculosis DNA amplification.Drug susceptibility and mutation sites of drug resistance genes were analyzed in MTB-L.Results Ⅰ-dentification of MTB-L:after the positive strain has been cultured,the colonies have the characteristics of "fried egg sample","particle-like" and " filament-like".MTB confirmed by tubercle DNA amplification experiments.Isolation rate:after cultured by MGIT 960 and Modified 92-3TB-L medium,the positive rate of single bacterial type was 50.90％,(113/222) the positive rate of both bacterial type and L type was 15.32％ (34/222),and the positive rate of MTB-L type was 2.25％ (5/222).Drug resistance:MTB-L was resistant to Streptomycin,Isoniazid,Rifampin,and Ethanol butylamine.No mutation was found in the drug resistance gene loci.Conclusions Clinical laboratory should routinely develop the culture of L-form of Mycobacterium tuberculosis bacteria,and increase the clinical attention to MTB-L.

Objective:To gain insight into the constitution of juvenile rheumatic diseases, treatment outcome and trends of rheumatic inpatients in past 12 years, and to improve awareness of juvenile rheumatic diseases.Methods:The clinical data of 5 950 patients in rheumatology department of the affiliated pediatric hospital of Fudan University (from 2005 to 2016) were analyzed retrospectively, and the chi-square test was used to compare and analyze the incidence.Results:Disease changes: ① The top three rheumatic diseases were Kawasaki disease (KD) (44.3%), Henoch-schoniein purpura (HSP) (35.4%), juvenile idiopathic arthritis (JIA)(9.6%). ② The number of all constitution of juvenile rheumatic diseases in hospital increased other than HSP. ③ The rheumatic diseases were increased from 17 to 37 kinds in the past 6 years. ④ The number of systemic lupus erythematosus (SLE) increased year by year (112/2 348 vs 197/3 602, χ2=1.41, P=0.235), as well as the severe SLE (35/112 vs 55/197, χ2=0.38, P=0.536). ⑤ The rate of rheumatic diseases complicated with macrophage activation (MAS) was 7.2‰(43/5 950). 12.9%(26/201) of systemic juvenile idiopathic arthritis(sJIA) were complicated with MAS, which was accounted for 60.5%(26/43) of total number of MAS in rheumatic diseases. In the last 6 years, there was a significant increase in the number of patients with MAS in patients with rheumatic diseases ( χ2=14.1, P<0.01) and sJIA( χ2=11.2, P<0.01). ⑥ 1.1%(64/5 950) of rheumatic diseases patients had lung lesions, juvenile dermatomyositis (JDM) accounted for 24.4%(20/82). In the last 6 years, the number of patients with lung lesions associated with rheumatic diseases increased significantly ( χ2=5.66, P=0.017). ⑦ The mortality rate of juvenile rheumatic diseases was only 3.7‰(22/5 950), and 45.5% occurred in SLE (10/22). The mortality rate of SLE decreased in last 6 years (5/112 vs 5/197, χ2=0.34, P=0.558). Conclusion:The constitution of juvenile rheumatic diseases in our center is decreasing for systemic vasculitis (KD, HSP), JIA, SLE, JDM in last 6 years. The annual total number of patients is relatively stable. But rare, difficult and critically illed cases increase year by year. Although SLE is still the primary cause of death in juvenile rheumatic diseases in recent 6 years, the mortality rate has decreased year by year.