OBJECTIVE To observe the anti-infection and anti-pyretic effect in vivo of Minor Decoction of Bupleurum(Xiaochaihu Tang).METHODS The Streptococcus pneumoniae infection model in mice and endotoxin-induced feverish model in rats were prepared,Minor Decoction of Bupleurum was used to treat the experimental animals vs Baphicacanthus cusia Bremek.(Isatis tinctoria)granule in control unimals to observe death rate of infected animal and the changes in temperature of feverish animals.RESULTS Minor Decoction of Bupleurum had better effect on decreasing death rate of S.pneumoniae infected animal and on lowering the temperature of endotoxin-induced feverish animal,P

Objective To observe the therapeutic effect and security of Thermal Moxibustion combined with Yougui Pills in the treatment of premature ovary failure(POF) of spleen and kidney Yang-deficiency.Methods One hundred and forty-two patients with POF of spleen and kidney Yang-deficiency were randomly divided into the treatment group and control group,71 cases in each group.The observation group were treated with Thermal Moxibustion combined with Yougui Pills,while the control group took oral Yougui Pills,12 weeks as a course of treatment.After treatment,the clinical therapeutic effects,K upperman score,related hormones levels(E2,FSH,LH) and occurrence rate of adverse reactions were compared between the two groups.Results The total efficacy rate of the observation group was 95.77％,which of the control group was 77.46％,and the observation group was significantly better than the control group(P＜0.05);there were significant improvement in the Kupperman score and levels of related hormones after treatment in the two groups(P＜0.05),but the improvement degree of the observation group was superior to the control group(P＜0.05).Conclusion Thermal Moxibustion combined with Yougui Pills can alleviate the clinical symptoms with high safety and higher clinical application value

Objective To improve the diagnosis of amniotic bands syndrome with ultrasound. Methods All pregnant women underwent ultrasonographic examination to observe the intra amniotic band like structures. Results Among 8771 women undergone ultrasound examination, 60 were found with intra amniotic band structure. Seven cases were diagnosed as amniotic bands syndrome and 53 intra amniotic band like structures included extra amniotic pregnancy(7), amniotic sheets(4), incomplete uterine septum(24), fetal nuchal cystic hygroma(2), fetal nuchal and trunk edema(11) and disclosed extra embryo sac(5). Conclusions Not all the intra amniotic band like structures is amniotic band. But any abnormal structures detected in early stage should be closely followed up.

Objective To identify clinicopathologic factors which predict survival following hepatectomy in HBV-related cirrhotic patients with early hepatocellular carcinoma (HCC).Methods A database was used to identify patients with histologically confirmed early HCC (≤5 cm,no nodal involvement,metastases,or major vascular invasion) who underwent surgical resection (excluding ablation or transplantation).Among 20 700 patients with HCC who were diagnosed at the Eastern Hepatobiliary Surgery Hospital from April 2005 to November 2010,537 (2.6％) patients with early HCC were studied retrospectively.Prognostic factors were evaluated using the Kaplan-Meier curves,Cox proportional hazards models and the receiver operating characteristic (ROC) curves.Results The study included 537 patients.The median tumor size was 2.9 cm,and 33％ of patients had tumors ≤2 cm.Most HCC lesions were solitary (63％) and had no evidence of vascular invasion (64％).Following surgery,the overall median and 5-year survival were 45 months and 33％ respectively.After adjusting for demographic factors and histological grade,tumor size ＞2 cm (hazard ratio [HR]:1.56),multifocal tumors (HR:1.34),and vascular invasion (HR:2.03) remained independent predictors of poor survival (all P ＜ 0.05).Based on these findings,a prognostic scoring system was developed that allotted 1 point each for these factors.Patients with early HCC could be stratified into 4 distinct prognostic groups (median and 5-year survival,respectively):0 points (97 months,96％),1 point (85 months,76％),2 points (76 months,54％),3 points (56 months,39％) (P ＜0.01).Conclusions The present study emphasized the importance of pathologic staging even in patients with small HCC.Anatomical resection of HCC should be the preferred surgical procedure in cirrhotic patients.

Objective To investigate the effects of heme oxygenase-1 (HO-1) preconditioning on oxidative damage in alveolar epithelial type Ⅱ (AE- Ⅱ) cells in rats. Methods The primarily cultured AE- Ⅱ cells isolated from male SD rats were randomly assigned to one of 6 groups (n = 8 each): control group (group C),H2O2 group and 4 different concentrations of HO-1 preconditioning group (group H1-4). The cells were continuously incubated for 5 h in group C. H2O2 0.5 mmol/L was added and the cells were incubated for3 h in group H2O2.In group H1-4, HO-1 0.01, 0.10, 1.00 and 10.00 μmol/L were added and the cells were incubated for2 h, then H2O2 0.5 mmol/L was added and the cells were incubated for 3 h. After the end of incubation, the cell morphology was observed under inverted phase contrast microscope, the AE- Ⅱ cells were counted, and the cell viability was determincd. Results The most AE- Ⅱ cells were adherent and round, had homogeneous cytoplasm, and the cytoplasm contains granular materials in group C and H2-4, while the vacuoles appeared in AE- Ⅱ cells and the cell debris appearecd in the supernatant in group H2 O2 and H1 . Compared with group C, the cell count and cell viability were significantly decreased in group H2 O2 and H1 (P ＜ 0.05). Compared with group H2 O2 and H1, the cell count and cell viability were significantly increased in group H2-4 (P ＜ 0.05). There was no significant difference in the cell count and cell viability between group H2O2 and H1, and among group H2～4(P ＞ 0.05) .Conclusion Preconditioning with 0.10-10.00 μmol/L HO-1 can reduce oxidative damage in rat AE- Ⅱ cells.

BACKGROUND: Dendritic cells, the most potent antigen presenting cells known at present, have been extensively used in the immunotherapy as adjuvant. OBJECTIVE: The present study was to construct Der p 2 eukaryotic expression vector and validate its expression in the mouse bone marrow-derived dendritic cells. DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Respiratory Disease,Xinqiao Hospital, Third Military Medical University of Chinese PLA between May and December 2005. MATERIALS: C57BL/6 mice were included. Plambd-Der p 2 was the product of Heska Company, USA.pCI-neo plasmid was provided by the Institute of Respiratory Disease, Xinqiao Hospital, Third Military Medical University of Chinese PLA.METHODS: Mouse bone marrow-derived dendritic cells were in vitro isolated and cultured.Complete Der p 2 cDNA was spliced from prokaryotic expression vector plambd-Der p 2, and then cloned into eukaryotic expression vector pCI-neo (pCI-neo-Der p 2).The positive recombinants pCl-neo-Der p 2 transfected into dendritic cells.Non-transfected and blank vector pCI-neo-transfected dendritic cells were used as controls. MAIN OUTCOME MEASURES: ①Identification of pCI-neo-Der p 2 recombinant plasmid.②Detection of Der p 2 mRNA and protein expression by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot techniques. RESULTS: Sequencing results showed Der p 2 cDNA in pCI-neo-Der p 2 was in coincidence with the sequence registrated in Gene Bank.RT-PCR and Western Blot results showed that expression of Der p 2 mRNA and protein could be detectable in the pCI-neo-Der p 2-transfected dendritic cells. CONCLUSION: The Der p 2 cDNA was successfully constructed into the eukaryotic expression vector, and Der p 2 gene and protein could be expressed efficiently in dendritic cells.

BACKGROUND: Techniques in the focal cerebral ischemia model in mice have been well developed in some foreign countries. However, rats are commonly used in cerebral ischemia studies due to resource availability in China.OBJECTIVE: To establish reliable and reproducible focal cerebral ischemia reperfusion model in mouse to explore the molecular mechanism of cerebral ischemia reperfusion process in genetic level.DESIGN: Randomized controlled study.SETTING: Fundamental Medical Research Institute of Chengde Medical College. MATERIALS: The experiment was carried out in Fundamental Medical Research Institute of Chengde Medical College from September 2002 to May 2003. Totally 30 Kunming mice (25 to 30 g), provided by Experimental Animal Center, Chengde Medical College, were randomly grouped as follows: control group, 6, 3-hour occlusion and 18-hour, 24-hour reperfusion, with 6 mice in each group respectively.INTERVENTIONS: Nylon thread (qb0.128 mm) was soaked in paraffin wax and inserted into internal carotid artery in mouse to reversibly occlude the middle cerebral at the right side after 3 hours, then was removed, and reperfusion was performed in 18, 24 hours, respectively.MAIN OUTCOME MEASURES: The infarct volume was observed and neurological deficit scored was analysis.of mice in operation group was shown as head turning to left, adduction of left forelimb, internal rotation of shoulder, decrease of muscular tension of left forelimb, unable to straighten completely during tail lifted. At independent activity, mice turned to left as the left hidlegs as the center of a circle; under the abdomen, left forelimb was crossed with right forelimb as scissor shape. Palsy of left limbs was observed obviously, especially forelimb.tion was shown clearly with TTC. The infarct volumes were (13±2.3) mm3 and (16±4.5) mm3 for 18 hours and 24 hours reperfusion and 3-hour ischemia, respectively. Pathological changes were observed under light microscope after ischemia. The infarct volume increased as the increasing of reperfusion time, including 3-hour blocking and 18-hour and 24-hour reperfusion.CONCLUSION: Using suture method to establish focal cerebral ischemia reperfusion model in mice, it is less invasion and trauma without requiring skull opening. Ischemia position is relatively fixed, thus, ischemia reperfusion time can be controlled precisely. It is also an ideal animal model to study pathological changes in cerebral vascular diseases and an efficient tool to study the therapeutic effect of medicine after surgery.

To investigate the neuroprotective of ligustilide (LIG) against glutamate-induced apoptosis of PC12 cells, cell viability were examined by MTT assay. Flow cytometry was applied to assay cell apoptosis rate. Intracellular calcium concentration was measured by using fluorescent dye Fluo-3/AM. Cytochrome C (Cyt C), Caspase-3, Bax and Bcl-2 protein expression were assayed by western blot. The results showed that glutamate is cytotoxic with an inhibitory concentration 50 (ID50) of 15 mmol · L(-1). Pretreatment with LIG (1, 5, 15 μmol · L(-1)) significantly improved cell viability. The apoptosis rate in glutamate-induced PC12 cells was 13.39%, and decreased in the presence of LIG (1, 5, 15 μmol · L(-1)) by 9.06%, 6.48%, 3.82%, separately. Extracellular accumulation of Ca2+ induced by glutamate were significantly reduced by LIG. The results of western blot manifested that pretreatment LIG could decrease the release of Cyt C from mitochondria, down-regulate Caspase-3 protein expression and up-regulate Bcl-2/Bax ratio, thereby protects PC12 cells from apoptosis. In summary, LIG had protective effect on glutamate-induced apoptosis in PC12 cells through attenuating the increase in intracellular Ca2+ concentration, and inhibiting the release of Cyt C from mitochondria to cytoplasm.

Objective:To investigate the cytokine profile expressed by CIK cells.Methods:CIK cells were induced from peripheral blood mononuclear cells in the presence of IFN-?,IL-1?,IL-2 and mAb against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis.Cytokine expression profiles were determined at mRNA level by semi-quantitative RT-PCR.Results:The percentage of CD3~+CD56~+ positive cells reached up to or higher than 50%.CIK cells couldn't express IFN-?、IFN-? and IFN-? under this kind of condition. But they could express IFNs principally on day 10 to 20. The level of ILs went down after steadily expressed on day 6 to 10(or 20).TNF-? and TRAIL constitutively expressed during the culture period in vitro. TNF-? maintained high expression on day 20.TGF-?_1 mainly expressed on day 30.Conclusion:CIK cells can widely express various kinds of cytokines in vitro.

Objective To compare the differences and characteristics of the dose distribution of the two optimization methods in the three dimensional brachytherapy,and provide the basis for clinieal treatment.Methods Excel 2007 was used to generate random number.And a total of 21 patients of cervical cancer were selected from those who have completed the treatment.Inverse simulated annealing optimization (IPSA) plans were designed for graphical optimization (GO) plans.The dose volume histogram (DVH) parameters of the targets (V100％,V150％) and the organs (D1 cm3,D2cm3) of the two methods were analyzed.Results The targets dose of both plans could meet the prescription requirements.There was no statistically significant difference in the dose parameters of all targets (P ＞ 0.05).The closes of D1 cm3 and D2cm3 in the bladder of IPSA plan were significantly lower than that of the GO plan (t =3.596,3.490,P ＜ 0.05).There was no statistically significant difference in the dose parameters of rectum (P ＞ 0.05).Conclusions For cervix brachytherapy,the GO and IPSA have no effect on targets dose,but IPSA optimization can reduce the maximum dose of bladder.