This study examined the roles of HLA-G in the pathogenesis, development and immune tolerance of hemangioma. From 2000 to 2007, 52 paraffin-embedded specimens (26 from males and 26 from females) of skin capillary hemangioma and 7 samples of adjacent normal skin tissues were collected. Four fresh specimens of hemangioma were also harvested. All samples were HE-stained and proliferative cell nuclear antigen (PCNA) was immunohistochemically detected by using SP method. The samples were classified into proliferative group and degenerative group according to the Mulliken criteria and the expression pattern of PCNA. SP method and quantum dots double staining were applied to detect the expression of HLA-G and PCNA in hemangioma and normal tissue samples. The expression of HLA-G was detected by RT-PCR. The results showed that among the 52 samples of hemangioma, 29 were of proliferative type and 23 degenerative type, and of the four fresh samples of hemangioma, 2 were of proliferative type and 2 degenerative type. SP method results showed that HLA-G was expressed in both proliferative and degenerative hemangioma, but not in normal tissues. The quantum dots double staining exhibited that HLA-G expression was significantly higher in proliferative group than in degenerative (P<0.05) and normal groups (P<0.05), but there was no statistically significant difference between the latter two groups (P>0.05). RT-PCR revealed that HLA-G was transcribed in both the proliferative and degenerative hemangioma tissues, but not in normal tissues. We are led to conclude that the elevated expression of HLA-G in proliferative hemangioma cells may lead to immune tolerance, which allows cells to escape immune surveillance and proliferate. On the other hand, the lower expression of HLA-G in degenerative hemangioma may result in immune cells-induced degeneration of hemangioma.

Objective Benign prostatic hyperplasia ( BPH) is one of common diseases in aged males , and searching for new therapeutic drugs to BPH has been a research hotspot in recent years .This article was to study the inhibitory effect of eupatorium ja-ponicum thunb and foeniculum vulgare extract ( EFE) on benign prostatic hyperplasia in rats and its possible mechanism . Methods 48 male rats were randomly divided into 6 groups:normal control group without any treatment , model group of BPH treated with subcu-taneous injection of testosterone propionate , positive control group of BPH treated with dutasteride , high, middle and low dosage groups according to different EFE dosage (156 mg/kg, 234 mg/kg and 312 mg/kg).45 days after the treatment, the rats were sacrificed for measurement of the prostate glandular wet weight , the index of prostate gland ( PI ) , the morphological changes of prostate gland by light microscopy and the content of sex hormone . Results The prostate wet weight and PI decreased after EFE treatment for 45 days compared with the BPH model group(P<0.01 ).The hyperplastic glandular epithelium papilla waned and even disappeared in three EFE groups under the light microscope , and the epithelial cells became cubical or flat .High dosage EFE group (312 mg/kg) has simi-lar efficacy to dutasteride group .EFE significantly reduced serum testosterone content , dihydrotestosterone content and T/E2 ratio( P<0.05 ). Conclusion EFE can significantly inhibit prostatic hyperplasia in rats , and its mechanism is related to the decrease of the contents of serum testosterone and dihydrotestosterone as well as T/E2 ratio.

Objective To observe the Klotho expression in kidneys and renal tubular epithelial cells apoptosis in spontaneously hypertensive rats (SHRs) and the effects of cordyceps sinensis (CS), in order to study the mechanism of protective effects of CS on renal tubular cells apoptosis in hypertensive renal damage. Methods Twenty 22-week-old male SHRs were control group. After 8 weeks, the levels of 24 hours urinary protein (Upre), urinary N-acetyl-β-D-glucosaminidase (NAG), serum creatinine (Scr), blood urea nitrogen (BUN) and renal pathological changes were detected; the mRNA expression of Klotho, 053 and 021 was detected by RT-PCR; the protein expression of Klotho, 053, 021 and cleaved-caspase-3 was tested by Western blotting. TUNEL assay was applied to evaluate the renal tubular cell apoptosis. Results As compared to SHR group, the levels of 24 h urinary protein content [(52.16±29.3) mg, (49.97±32.5) mg, (54.67±30.09) mg vs (96.52±36.94) mg], urinary NAG [(44.13±9.11), (42.75±8.33), (41.96±7.88) U/L vs (54.07±6.57) U/L], Sct [(45.25±9.55), (43.76±8.65), (45.18±7.28) μmol/L vs (53.84±10.21) μmol/L]and BUN [(8.25±1.03), (8.40±1.58), (8.32±0.98) mmol/L vs (8.91±1.24) mmol/L]were decreased (all P<0.05), renal pathological changes were relieved, the levels of Klotho expression were up-regulated and the levels of p53 and p21 expression and cleaved-caspase-3 protein expression were down-regulated (all P<0.01), tubular cell apoptosis was decreased [7.56%±0.52%, 7.93%±0.37%, 7.37%±0.36% vs 13.32%±0.64%, P<0.01] in CS, Los and CS+Los group. Conclusions Klotho, p53 and p21 play important roles in renal tubular cells apoptosis in hypertensive renal damage. CS can up-regulate Klotho expression, down-regulate p53 and p21 expression and decrease the cleaved-caspase-3 expression and tubular cell apoptosis to ameliorate the hypertensive renal damage.

OBJECTIVETo evaluate the potential usefulness of a multivariable model, established mainly upon peripheral T-cell subsets, Th1/Th2, T-bet and GATA-3 gene expressions as well as TCM and Western medical diagnostic criteria, in predicting the selection of immunosuppressive therapy (IST) or androgens in treating patients with aplastic anemia (AA).

METHODSPeripheral blood T cell subsets in 85 patients with AA were serially analyzed by flow cytometry before and after treatment, and their T-bet and GATA-3 gene expressions were assessed meantime by Real-time PCR. Then analysis of Logistic regression equation and ROC curves were performed based on the cases responding to IST or androgens.

RESULTS(1) According to the logistic equation and ROC curve of SPSS, setting the false positive rate as 0.10, the P value was 0.832. When P> or =0.832, patients were judged in the immunosuppressive dominant state, IST should be applied; when P<0.832, it means patients in the bone marrow failure dominant state, androgens should be added. (2) A novel theory is raised by the above-mentioned analysis, which indicated that the genesis and development process of AA could be divided in 2 stages, the abnormal immune dominant stage and the bone marrow failure dominant stage. For treatment of patients in the two stages, IST and androgens is the preference respectively.

CONCLUSIONThe multivariable model could be used for indicating which stage the AA patient is in, the abnormal immune stage or the bone marrow failure stage, and thus to guide the proper selection of IST or androgens in clinical practice.

Adult ; Aged ; Androgens ; therapeutic use ; Anemia, Aplastic ; drug therapy ; immunology ; Cyclosporine ; therapeutic use ; Female ; GATA3 Transcription Factor ; metabolism ; Humans ; Immunosuppressive Agents ; therapeutic use ; Logistic Models ; Male ; Middle Aged ; T-Box Domain Proteins ; metabolism ; T-Lymphocyte Subsets ; immunology ; Young Adult

Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P ＜0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P＞0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.

An iodine analysis instrument with sequential injection of urine samples was developed. A method for measurement of urine iodine was also developed by combining sequential injection and catalysis kinetics,and making use of catalysis facilitation of the iodine in the redox reaction of As~(3+) and Ce~(4+) . The sequential injection and stopped-flow stabilization determination were made possible by the program-controlled injector with controlled flow rate and the 16-hole program-controlled selection valves. The arsenic-cerium reaction with iodine-catalyzed at constant temperature state might proceed with the constant temperature flow cell. Using the syringes with program-controlled velocity,pushing and suction,Using the digital connected circuit and micro-iodine determination software,the reaction temperature of (32.0±0.1)℃,injection time of 45 s,stabilization time of 60 s,detection time of 20 s,injection volume of 400μL,linear range of 15 -600μg/L,detecting limitation of 5.01 μg/L(n=11) and recovery rate of 94. 1 % - 105. 1 % were obtained. With this method,the detecting result of the National Standard Reference (GBW09109 and GBW09110) materials was within a given standard range. Through this method,the detecting results had no significant differences comparing with those by standard method of National Health Service(P >0.05).

Objective To determine the effect of different serum insulin-like growth factor-Ⅰ(IGF-1)levels on mouse cancer.Methods A total of 120 male mice at 6 weeks of age(60 control mice and 60 LID mice)were subcutaneously injected colon tumor CT26 cell line.Each group was random divided into two subgroups respectively,every 10 mice of one subgroup were injected subcutaneously with growth hormone(GH)(1ms/kg)daily from the 10th,14th and18th days respectively until the 22nd days,and the other subgroup received saline injection.Results All mice treated with GH have higher level of IGF-1,compared with those treated with saline.High level of IGF-1 promoted the development of cachexia in these mice treated with GH from the 10th days.However,the level of IGF-1 has negative correlation with the cancer cachexia state for mice treated with GH from the 14th days.Conclusion Circulating IGF-1 and GH play an important role in tumor growth 4nd cachexia development in the early stage of cancer and can ameliorate the state of cachexia in the advanced stage.

Objective To explore the effects of application of probiotics on intestinal flora and related cytokines in infants with cholestatic liver disease.Methods (1) Eighty-four infants with cholestatic liver disease had been hospitalized from October 2010 to June 2011 in the First Hospital of Guangxi Medical University.The infants with cholestatic liver disease were randomly divided into the probiotic intervention group and the non-probiotic intervention group.Quantification of intestinal bacteria was detected by SYBR Green Ⅰ real-time fluorescent quantitative polymerase chain reaction,then the number of 3 kinds of bacteria before and after the treatment was compared.(2) The indices of liver function,blood ammonia,cholesterol were detected.The levels of serum transforming growth factor-β1,tumor necrosis factor-α(TNF-α)and interleukin-6 (IL-6) were measured by enzyme linked immunosorbent assay.Results (1) The number of bifidobacterium,lactobacillus and bifidobacterium/escherichia coli (B/E) were significantly increased (all P ＜ 0.01),whereas the number of escherichia coli was significantly decreased in the probiotic intervention group(P ＜ 0.05),however,there were no differences in the non-probiotic intervention group(all P ＞ 0.05).(2)The indices of total billirubin,direct billirubin,γ-Glutamyltransferase,total bile acid,alanine aminotransferase,blood ammonia,alkaline phosphatase were significantly improved after therapy in 2 groups (all P ＜ 0.05).The levels of TNF-α and IL-6 were significantly decreased in the probiotic intervention group (t--7.31,P =0.00;t =-2.90,P =0.01),but there were no differences in the non-probiotic intervention group.The level of BA was significantly decreased in the probiotic intervention group than the non probiotic intervention group (t =-8.37,P =0.00).(3) The B/E value were significantly inverse correlated with level of serum IL-6 (r =-0.796,P =0.01).Conclusions It may help to restore the intestinal flora and balance the immune function in infants with cholestatic liver disease after application of probiotics.

Objective To evaluate the effects of different doses of dexmedetomidine on median effective end-tidal concentration of sevoflurane (EC50) inhibiting responses to tracheal intubation in pediatric patients.Methods Sixty-seven ASA physical status Ⅰ or Ⅱ patients,aged 3-8 yr,with body weight not exceeding 150％ of the ideal weight,scheduled for elective surgery under general anesthesia,were randomly divided into 3 groups using a random number table:control group (group C,n =22) and different doses of dexmedetomidine groups (group D1,n =23 ; group D2,n =22).Before induction of anesthesia,dexmedetomidine 1.0 and 2.0 μg/kg was infused intravenously over 10 min followed by infusion at a rate of 0.5 and 1.0 μg·kg-1 ·h-1 in D1 and D2 groups,respectively.Anesthesia was induced with inhalation of 5 ％ sevoflurane.After eyelash reflex disappeared,the end-tidal sevoflurane concentration was adjusted to achieve the target concentration and maintained at this level for 15 min.Tracheal intubation was then performed and the response to intubation was scored.The initial end-tidal sevoflurane concentration was 3.5％,2.5％ and 1.5％ in C,D1 and D2 groups,respectively.Up-and-down sequential trial was used to determine the EC50.Each time the concentration of sevoflurane increased/decreased in the next patient depending on whether or not the response to intubation was positive.The positive response to intubation was defined as intubation score ＞ 1.The ratio of concentrations between the two consecutive patients was 1.2.The EC50 and 95％ confidence interval of sevoflurane were calculated.The development of adverse cardiovascular events was recorded after dexmedetomidine administration.Results The adverse cardiovascular events were not observed in D1 group.The incidence of hypotension and brachycardia was 14％ and 9％ in D2 group.The EC50 (95％ confidence interval) of sevoflurane was 3.54％ (3.39％-3.69％),2.37％ (2.24％-2.46％) and 1.41 ％ (1.37％-1.46 ％) in C,D1 and D2 groups,respectively.Compared with group C,the EC50 of sevoflurane was significantly decreased in D1 and D2 groups (P ＜ 0.01).Compared with group D1,the EC50 of sevoflurane was significantly decreased in group D2 (P ＜ 0.01).Conclusion The optimum dose of dexmedetomidine is 1.0 μg/kg (loading dose) and 0.5 μg· kg-1 · h-1 (maintenance dose) when combined with sevoflurane for induction of anesthesia in pediatric patients.