The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

With the development of tertiary treatment system,the higher quality of inter-hospital transport is the most critical factor for the prognosis of the critically ill pediatric patients,especially for pediatric patients whose condition is deteriorating rapidly.Improving the quality,creating the guideline,and establishing the network platform of inter-hospital will provide complete guarantee for inter-hospital transport of critically ill pediatric patients.With the development of information technology,the purpose of this study was to analyze the function of the network platform of inter-hospital transport in Hunan Children′s Hospital.

Objective To raise the awareness of the glandularis ureteritis. Methods Five cases with glandularis ureteritis confirmed by pathological examination from Jan 2001 to Jun 2012 were reviewed. The diagnosis was assessed on histopathologic studies in all. The clinical manifestations, the diagnosis and treatment were discussed. Results Resections of the affected ureter and ureteroplasty were undertaken for 2 patients. Ureteronephrectomy was performed for 2 patients because of serious hydronephrosis due to obstruction. Electrofulguration via-ureteroscopy was carried out for another patient. All patients have been well and without recurrence on following up for 15 to 152 months, with an average of 86.6 months. Conclusion Glandularis ureteritis is not frequently seen and the etiopathogenesis is not yet clear. Biopsy is very important for suspected neoplasm of ureter before performing nephrectomy.

BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear.
OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts.
METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected
serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse
transcription-PCR in al groups.
RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor
activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (P<0.01). These findings indicate that Chinese medicine Gukang prescription can modulate the expression of
core binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.

Objective To observe the changes of number and proportion of CD4+CD25+FOXP3+ cells in peripheral blood of active rheumatoid arthritis patients (RA),and explore the function of recombinant human TNFR Ⅱ-Fc on the CD4+CD25-FOXP3+ Treg cells.Methods ① Forty severe RA patients were selected,who were divided into the combined treatment group (TNFR Ⅱ-Fc+MTX) and MTX only group according to the principle of randomized,double-blind,parallel and placebo-controlled study.All patients were treated for 12 weeks.Flow cytometry was used to analyze and compare the expression ratio of CD4+CD25+FOXP3 +Treg cells of RA patients' peripheral blood.Forty healthy controls were selected for parallel comparison.② VAS,DAS28,HAQ average of the two groups at different periods were compared.Matched t test was used to examine the quantity data between the groups.Results ① The proportion of CD4+CD25+FOXP3+ cells in the peripheral blood of active rheumatoid arthritis patients was significantly lower than healthy group [ (5.4±1.4)％ vs ( 7.5±1.5 )％,P＜0.01 ].The proportion of CD4+CD25+ FOXP3+ Treg cells in combined treatment group was significantly higher [(7.0+1.2)％ vs (5.2±1.6)％,P＜0.01 ] after TNFR Ⅱ-Fc and MTX combination therapy for 12 weeks.The increase rate of CD4+CD25+FOXP3+Treg cells in combined treatment group was evidently more remarkable than MTX only group [ (7.0 ± 1.2)％ vs (5.6 ±0.7 )％,P＜0.01].② After 12 weeks treatment,the arerage scores of VAS,DAS28 and HAQ of the combined treatment group were better than MTX only group,and the difference was statistically significant (P＜0.01).Conclusion This study has shown that the healing effect of TNFR Ⅱ -Fc combined with MTX is better than MTX only.TNFR Ⅱ -Fc can restore and improve the proportion of CD4+CD25+FOXP3+ Treg cells in active RA patients,which is likely to be an important mechanism for the treatment of RA patients.

OBJECTIVETo investigate the changes in the circulating levels of Treg cells in patients with rheumatoid arthritis (RA) and their associations with the disease activity.

METHODSThe fraction of circulating CD4+CD25+FOXP3+ Treg cells in 40 active RA patients and 40 healthy controls were determined by flow cytometry. The serum levels of interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) were measured using enzyme-linked immunosorbent assay, and the expression of Foxp3 mRNA was detected with real-time PCR. The correlation of the changes in the fraction of Treg cells and the disease activity of RA was analyzed.

RESULTSRA patients showed a significantly lower level of circulating Treg cells than the control subjects [(5.36∓1.55)% vs (7.49∓1.46)%, P<0.01]. The expression of Foxp3 mRNA (P<0.01) and serum IL-10 level (P=0.000) were significantly lower, whereas TGF-β1 significantly higher (P=0.000) in RA patients than in the controls. Spearman analysis showed that serum level of IL-10 but not TGF-β1 was correlated to the fraction of Treg cells and Foxp3 mRNA expression, but the fraction of Treg cells was not correlated to such indices of disease activity as tender joint counts, swollen joint counts, visual analog scale, HAQ, disease activity score in 28 joints, ESR, or CRP, nor to RA self-antibodies (including RF and anti-CCP antibodies).

CONCLUSIONA lower fraction and dysfunction of circulating Treg cells might be involved in the pathologies of RA, and a higher disease activity is associated with a greater reduction of Treg cells.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; pathology ; physiopathology ; Blood Sedimentation ; CD4 Lymphocyte Count ; Case-Control Studies ; Female ; Flow Cytometry ; Forkhead Transcription Factors ; metabolism ; Humans ; Interleukin-10 ; blood ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; cytology ; Transforming Growth Factor beta1 ; blood

Objective To discuss the long term clinical effect of ileal orthotropic neobladder.Methods From 1991 to 1998,79 patients,mean age 55(41～75)years,male 74,female 6,were followed up.The serum creatinine and urea,electrolytes,blood routine,B ultrasonic scan of the neobladder residual urine and IVU or MRU of the patients were followed up.The max transverse diameter of renal pelvis and the max verticaI/level diameter of neobladder were measured in 5,10 to 14,15 years of postoperative when IVU or MRU.All results of different time were compared by the multiple comparisons.The local or distant cancer recurrence and the complications of the operation Were evaluated. Results Sixty-four cases,58 male,6 famle,were long term followed up:mean time was 167 (range,121～216)months.Seven cases died of other diseases.Seven cases had pelvic recarrence.Two cases had urethral recurrence.Three cases died of tumor metastasis.One case had ureter recurrence.Forty-eight patients were alive more than 10 years.The value of the serum creatinine,urea,electrolytes and bloods routine of the patients were normal after 5,10 to 14 and 15 vears postoperative (P＞0.05).The max transverse diameter of the renal pelvis in 5,10 to 14 and 15 years Dostoperative were 14.0 mm,14.1 mm and 13.7 mm,respectively,P＞0.05.The max vertical/level diameter of the neobladder in 5,10 to 14,15 years of postoperative were 110.4 mm/90.4 mm,111.5 mm/95.3mm and 127.0 mm/97.0 mm,respectively,P＞0.05.The residual urine of 5 cases was more than 50 ml and had not increased during follow up.Eight cases with neobladder stone were cured by the intracavitary lithothrypsis.Two cases with uretheral stricture were cured by the intracavitary therapy.Twelve cases of 14 cases with inguinal hernia were cured by reoperation,2 cases accepted conservative treatment.Only 17 cases had no complication involve of the cancer and the operation. Conclusion The upper urinary tract and neobladder of the ileal orthotopic neobladder could be stable for long time,the cure rate of tumor is satisfactory and the lifetime follow up is necessary.

Objective To investigate the expressions of Dapper1 in gastric carcinoma and elucidate its relationship with survivin and its role in tumor cell apoptosis. Methods Dapper1 mRNA was detected with RT-PCR using specimens from 30 cases of gastric carcinoma and the corresponding normal gastric mucosa.The pcDNA3.1-Dpr1 plasmid was transfected into SGC-7901 cells with LipofectamineTM 2000.The effect of upregulation of Dpr1 on SGC7901 cell apoptosis was determined by flow cytometry.The downregulation of survivin、Dvl-2 and β-catenin protein expression were detected by Western blot analysis.Results Downregulation of Dpr1 gene expression was observed in 17(57%)of 30 human gastric cancer and the downregulation was significantly correlated with the depth of invasion and the degree of differentiation (P＜0.05).Also,upregulation of Dpr1 mRNA and downregulation of survivin mRNA were detected after transfecting pcDNA3.1-Dpr1 plasmid in SGC7901 cells,which led to downregulation of survivin、Dvl-2、β-catenin protein and increase of the SGC7901 cell apoptosis rate from 2.89%to 13.96%.Conclusion Downregulation of Dp1l gene expression is common in human gastric carcinoma,and upregulation of Dpr1 results in significant inhibition of survivin expression which can induce apoptosis of SGC7901 cells.