Objective To investigate the risk factors and the immune functions for noeocomial palmonary fungal infection (PFI) in COPD patients.Methods The data of 55 cases of nosocomial PFI were analyzed,and examined the level of T Lymphocyte Subsets (TLS) in peripheral blood from 67 cases COPD,then compared with 54 healthy donors.Results Nosocomial pulmonary fungal infection Candida Albicans was ranked the first pathogen.CD3 and CD4 of COPD with PFI were significantly ower than COPD and healthy donors(P＜0.05).Conclusions We should improve the immunity of patients in the process of therapy.We should reasonable use of bread spectrum antibiotics and corticosteroids in order to reduce incidence of pulmonary fungal infection.

Objective To build rat orthotopic liver transplantation model and use this model as an experimental animal model for further research about liver transplantation.Methods Based on the classic two-cuff technique,a variety of skills were integrated,such as abdominal aorta perfusion first before ligation,suprahepatic inferior vena cava single continuous suture,preoperative and postoperative rehydration,hepatic portal vein and infrahepatic vena cava blocked by mosquito's haemostatic forceps.At last,the postoperative survival rates of 2 days and 2 weeks were recorded.Results Donors' operation time was (35±2) min,cuffing time was (7.0±1.5) min,receptors' operation time was (80±15) min,anhepatic phase was (21±3) min.The 2-day survival rate was 92 ％ (46/50) and 2-week survival rate was 86 ％ (43/50).Conclusion To some extent,the improved methods can not only reduce the difficulty of orthotopic liver transplantation operation,but also to create a uniform,stable and reliable orthotopic liver transplantation model.

Objective To observe the effect of BSD 2000 deep thermotherapy plus chemotherapy in treatment of malignant seroperitoneum of advanced epithelial ovarian cancer patients with drug resistance.Methods 36 advanced epithelial ovarian cancer patients with malignant seroperitoneum for drug resistance were randomly divided into two groups,trial group (18 cases) and control group (18 cases).Cases in trial group were treated with BSD 2000 deep thermotherapy plus GT regimen (gemcitabine 1 000 mg/m2 iv d1,d8,taxinol 80 mg/m2 ip d1,d8.28 days for a cycle),while control group with GT regimen alone.Effect,survival time (median) toxicity and Karnofsky score were evaluated after 2 cycles.Results Response rate (RR) was strongly higher in trial group compared with control group [55.6 ％ (10/18) vs 22.2 ％ (4/18),P ＜ 0.05],the same to disease control rate (DCR),but there was not significant difference between two groups (P ＞ 0.05).The improvement rate of Karnofsky score in trial group was higher than that in control group,which had no significance (P ＞ 0.05).The toxicity were similar in both groups,which had no stage 3 to 4 side-effect.The differences of survival time (median) and survival rate had no statistical significance between two groups (P ＞ 0.05).Conclusion It is useful to eliminate seroperitoneum,improve quality of life and decrease the toxicity for the regimen of BSD 2000 deep thermotherapy plus chemotherapy in treatment of malignant seroperitoneum of advanced epithelial ovarian cancer patients with drug resistance.

Objective To investigate the response of mesenchymal stem cells in mice to mediumdose X-ray irradiation in vitro.Methods The mouse mesenchymal stem cell line C3H10T1/2 was submitted to 3.5 Gy X-ray irradiation.Hoechst33258 staining of adherent cells and Annexin V-FITC staining and flow cytometry analysis of suspension cells were performed respectively to assess cellular apoptosis at 3,6,12,24,48,72 h and 1 week after irradiation.SA-β-gal staining was performed to analyze the cellular senescence at 24,48,72 h and 1 week after irradiation.The mRNA level of both Fas with its ligand FasL and p53 with its downstream target p21 WAF1 were measured by Real-Time PCR analysis.The expression of Fas protein was determined by immunofluorescence staining.Results An increased apoptosis was observed at 3 h after irradiation with apoptosis rate 11.72％ ± 1.61％ ( t =9.01,P ＜0.01 ),the apoptosis rate reached the peak level at 12 h 20.52％ ± 1.96％ (t =16.27,P ＜ 0.01 ),and then declined progressively to normal level at 48 h 4.93％ ±0.46％ (t =2.26,P ＞0.05).The SA-β-gal positive rate of post-radiation cells at 72 h was 53.33％ ± 5.62％,significantly higher than that of normal control 3.24％ ± 0.39％ (t =17.77,P ＜ 0.01 ).The level of Fas,FasL mRNA was found to be elevated 3 h after irradiation with a peak at 12 h,and no differences were found l week later.The level of Fas protein was observed to reach the peak at 12 h after irradiation.The occurrence of peak level of Fas/FasL mRNA and protein was consistent with that of apoptosis of C3H10T1/2 cell.A transient up-regulation of p53,p21 WAF1 mRNA expression was found at 12 h after irradiation followed by a significant increase later at 72 h after irradiation.The occurrence of the two peaks of p53,p21WAF1 mRNA expression were coincident with that of cellular apoptosis and senescence,respectively.The levels of p53,p21WAF1 mRNA in senescence group were significantly higher than those of apoptosis group ( t =17.85,13.36,P ＜ 0.01 ).Conclusions The MSC cell line C3H10T1/2 was sensitive to medium-dose X-ray irradiation.Cell apoptosis occurred immediately after irradiation and cellular senescence happened at advanced stage.Both Fas/FasL and p53/p21 WAF1 signal pathway mediate the injury of C3H10T1/2 cell to medium-dose X-ray irradiation exposure.

Objective To evaluate the quality coherence of commercial Angelicae Dahuricae Radix (Baizhi) decoction pieces. Methods The samples of 12 batches of commercial decoction pieces, 1 batch of sulphur fumigated Baizhi, and 1 batch of naturally dried Baizhi were collected. HPLC method was used to determine the fingerprints and the contents of imperatorin and isoimperatorin by C18 column (4.6 mm×250 mm, 5 μm) with a gradient mobile phase of acetonitrile-water solution system at the flow rate of 1.0 mL/min. The column temperature was set at 35 ℃, and the max plot of detection wavelength was in 210-800 nm. Similarity calculation was used to analyze the data. Results The HPLC fingerprint analysis method was established with 12 common peaks. The similarities of 12 batches of commercial decoction pieces of Baizhi were 0.840-0.973. Their similarities compared with the sulphur fumigated Baizhi were 0.672-0.908. Compared with naturally dried Baizhi, the similarities of fingerprint were 0.536-0.684. The contents of imperatorin and isoimperatorin in 12 batches of commercial decoction pieces were 0.035%-0.140% and 0.028%-0.069%, respectively. Conclusion The quality of Baizhi decoction pieces was consistent. It can be speculated that Baizhi decoction pieces were processed with sulphur fumigation.

Object: To investigate the effects of Huaiqihuang Granule, a compound Chinese herbal medicine, on expressions of nephrin and podocin of slit diaphragm of glomerular podocytes in rats with adriamycin-induced nephrosis and to explore the mechanism in reducing the proteinuria. Methods: Twenty SD rats were randomly divided into five groups: control group, model group, glucocorticoid group, Huaiqihuang Granule group and Huaiqihuang Granule plus glucocorticoid group. The 24-hour urine was collected 7, 14, 21 and 28 days after adriamycin injection respectively to measure 24-hour urinary protein, and all rats were sacrificed after 28-day treatment. Pathological changes in renal tissues were observed under a light microscope and an electron microscope. Expressions of nephrin and podocin mRNAs in renal cortex were determined by real-time polymerase chain reaction, and protein levels of nephrin and podocin were detected by Western blotting. Results: (1) In the model group and the treatment groups, the level of urinary protein increased significantly from the 14th day. (2) Under the light microscope, inflammatory cells and slight fibroplasia were found in renal interstitium of the model group, but there were less inflammatory cells in renal interstitium in the intervention groups than in the model group. Under the electron microscope, 29 days after adriamycin injection, extensive fusion of foot processes was observed. (3) The expressions of nephrin and podocin were higher in treatment groups than in the model group. (4) Proteinuria level was negatively correlated with the expressions of nephrin mRNA and nephrin and podocin proteins. Conclusion: The above results indicate that Huaiqihuang Granule can maintain the integrity of the slid diaphragram in podocyte, alleviate the lesion of glomerular filtration membrane, and decrease the proteinuria by up-regulating the expressions of nephrin and podocin. Huaiqihuang Granule plus glucocorticoid maybe has better effects than glucocorticoid alone.

Objective The purpose of this study was to evaluate effects of gonadotropin-releasing hormone analogs (GnRHa) on improvement of predicted adult height (PAH) in girls with idiopathic central precocious puberty (ICPP) initiating treatment at age of eight years or over and analyze its related factors.Methods Forty-nine girls with ICPP aged eight years and over were treated with GnRHa for ( 17.4 ±5.6) months initiating at age of (9.3 ± 0.6) years during 2005 and 2008.They were followed-up every three to six months to evaluate its effects on growth velocity (GV), height standard deviation score for chronological age (HtSDS-CA), height SDS for bone age (HtSDS-BA), PAH, and so on, as well as their related factors.Results PAH increased after GnRHa treatment, from ( 157.7 ± 4.5) cm six months after treatment to ( 159.2 ±4.4) cm 12 months after treatment, and to ( 160.8 ±5.0) cm by the end of treatment from ( 155.5 ±5.1 ) cm at its initiation (P ＜0.01 ).There was significant difference in PAH by the end of treatment and target height (TH) [( 157.6 ±3.4) cm].HtSDS-BA increased since six months after GnRHa treatment, from ( -0.64 ±0.68) six months to ( -0.52 ±0.70) 12 months after it and ( -0.36 ±0.68)by the its end, all significantly different from ( - 0.94 ± 0.68 ) that at the initiation of treatment ( P ＜0.01 ).There was no significant difference in HtSDS-CA before and after treatment.GV decreased from the initiation (6.3 ± 1.3) cm/yr to (4.8 ± 1.5) cm/yr by the end of treatment ( P ＜ 0.01 ).No significant association between PAH and age of the initiation of treatment and between PAH and age by the end of treatment were found (P ＞ 0.05 ).But, PAH reversely correlated with bone age, and positively correlated with HtSDS-CA, HtSDS-BA, GV, TH and length of treatment ( P ＜ 0.01 ).Conclusions GnRHa can remarkably improves PAH in girl with ICPP.PAH mainly correlates with HtSDS-BA, TH and GV at the initiation and the end of treatment.

[ABSTRACT]AIM:Toexploretheroleofkisspeptin/GPR54signalingpathwayinthepathogenesisofprecocious puberty based on female precocious puberty rat model induced by the single dose of danazol .METHODS:Female SD rats aged 3 d were randomly divided into normal group , vehicle group and model group .On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture .All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development .The levels of E2 , FSH and LH in peripheral blood were measured by ELISA .The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR.The kisspeptin expression in rat hypothalamus was observed by immunofluo-rescence .RESULTS:The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group .On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group .On days 25 and 30, the ovarian mor-phological development in the model rats was significantly earlier than that in normal group .On day 25, the mRNA expres-sion of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats signifi -cantly increased compared with normal group and vehicle group .On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group .On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group .The mRNA expression of GPR54 had no obvious difference among all groups .CONCLUSION:The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty , suggesting that kisspeptin is an initiating factor for precocious puberty .Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty .

Tissue-engineering bone with porous ,betatricalcium phosphate (3-TCP) ceramic and autologous bone marrow mesenchymal stem cells (MSC) was constructed and the effect of this composite on healing of segmental bone defects was investigated. 10-15 ml bone marrow aspirates were harvested from the iliac crest of sheep, and enriched for MSC by density gradient centrifugation over a Percoll cushion (1. 073 g/ml). After cultured and proliferated, tissue-engineering bones were constructed with these,cellS seeded onto porous f-TCP, and then the constructs were implanted in 8 sheep left metatarsus defect (25 mm in length) as experimental group. Porous ,-TCP only were implanted to bridge same size and position defects in 8 sheep as control group, and 25 mm segmental bone defects of left metatarsus were left empty in 4 sheep as blank group. Sheep were sacrificed on the 6th, 12th, and 24th week postoperatively and the implants samples were examined by radiograph, histology, and biomechanical test. The 4 sheep in blank group were sacrificed on the 24th week postoperatively. The results showed that new bone tissues were observed either radiographic or histologically at the defects of experimental group as early as 6th week postoperatively, but not in control group, and osteoid tissue, woven bone and lamellar bone occurred earlier than in control group in which the bone defects were repaired in "creep substitution" way, because of the new bone formed in direct manner without progression through a cartilaginous intermediate. At the 24th week, radiographs and biomechanical test revealed an almost complete repair of the defect of experimental group, only partly in control group. The bone defects in blank group were non-healing at the 24th week. It was concluded that engineering bones constructed with porous -TCP and autologous MSC were capable of repairing segmental bone defects in sheep metatarsus beyond "creep substitution" way and making it healed earlier. Porous ,-TCP being constituted with autologous MSC may be a good option in healing critical segmental bone defects in clinical practice and provide insight for future clinical repair of segmental defect.
Bone Marrow Cells/cytology ; Calcium Phosphates/*pharmacology ; Cells, Cultured ; Fractures, Bone/*therapy ; Implants, Experimental ; Mesenchymal Stem Cells/*cytology ; Metatarsus/*injuries ; Porosity ; Sheep ; Tissue Engineering

Objective To construct recombinant adenovirus Ad/MDC-VP1 and investigate its im-muno-boosting effect of the mice primed with the experimental DNA vaccine against Coxsackievirus infection. Methods The recombinant adenovirus Ad/MDC-VP1 was constructed and packaged. The Western blot analysis was used to verify the target protein. BALB/c mice were divided into four groups: Ad/MDC-VP1 group, pcDNA3/MDC-VP1 group, pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group and PBS group. The mice in each group were immunized intramuscularly. The titers of serum IgG and neutralizing antibody were tested by ELISA and trace neutralization assay, respectively. The lymphocytes proliferation activity and specific CTL cytotoxic activity were tested by CCK-8 assay. The mice in each group were challenged with le-thal dose of Coxsackievirus, and the assay of the serum virus titers and the observation of protection efficacy against Coxsackievirus infection were carried out. Results The recombinant adenovirus Ad/MDC-VP1 was successfully constructed and the target protein was expressed. It was observed that the titers of CVB3 VP1 specific antibody, lymphocyte stimulation index, CTL cytotoxicity activities and protection rate of the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost group were much higher than those of the rest groups( P < 0.05), and the titer of serum virus was lower after CVB3 challenged ( P < 0.05 ). Conclusion Both the cellular and humoral immune responses in mice could been significantly enhanced by the pcDNA3/MDC-VP1 prime-Ad/MDC-VP1 boost strategy.