Objective To investigate the construction of compound membrane with corneal stromal cells and collagen-chitosan by tissue engineering technique and its biocompatibility.Methods Rabbit and human corneal stromal cells were separated and seeded into collagen-chitosan membrane.The compound membrane was transplanted into rabbit corneal stroma.Then the growth condition of keratocytes,the effect on normal keratocytes and degradation of compound membrane were detected by corneal confocal microscope,anterior OCT and histological and immunohistochemical methods ex vivo 1,2,4 weeks after grafting.Results The rabbit and human corneal stromal cells grown well in collagen-chitosan scaffold.The compound membrane degradated gradually after grafting.There was no necrosis and dissolvation.Corneal epithelium,stroma and endothelial cells were all normal.Conclusion Collagen-chitosan can be used as a biological scaffold for construction of corneal stroma.Corneal confocal microscopy and anterior OCT are new methods to observe the biological activity of constructed corneal stroma.

Objective To investigate effect of folic acid combined with xin funing on CRP, HGF, IL-2,TNF-αof patients with cervical cancer caused by human papillomavirus.Methods 80 cases of cervical cancer patients were randomly divided into control group, 40 cases in the control group were given conventional chemotherapy, 40 cases in the experimental group were on the base of the control with folic acid combined with xin funing.CRP, HGF, TNF-α, IL-2 and T lymphocyte subsets were compared before and after the treatment.Results Compared with the control group, the serum CRP, HGF and TNF-αof the experiment group were lower(P<0.05), IL-2 levels was higher (P<0.05), CD4 +and CD4 +/CD8 +level were higher(P<0.05), level of CD8 +was lower(P<0.05) and the clinical effective rate were higher(P<0.05).Conclusion Folic acid combined with Xin Funing has important significance for the treatment of patients with cervical cancer.It is speculated that the mechanism may be to reduce the level of serum CRP and HGF in patients with cervical cancer, and to increase the level of IL-2, and to regulate immune cells.

OBJECTIVE:To study the effects and mechanism of rapamycin on invasion and metastasis of cervical cancer HeLa cell. METHODS:HeLa cells were divided into control group and rapamycin low-dose,medium-dose and high-dose groups (10, 30,100 nmol/L). After treated for 48 h,cell viability was measured by MTT assay,and inhibitory rate was calculated;migration and invasion of cell was tested by Transwell assay. The expression of matrix metalloproteinase 2(MMP-2),MMP-9,Vimentin and E-cadherin,and phosphorylation of protein kinase B (Akt),mammalian target of rapamycin (mTOR) were detected by Western blot. RESULTS:Compared with control group,the inhibition rate of cell viability was increased in rapamycin groups(P<0.01);the number of invasion and metastasis cells decreased(P<0.01);the expression of MMP-2,MMP-9 and Vimentin were decreased (P<0.01 or P<0.05);the expression of E-cadherin was enhanced(P<0.01 or P<0.05);the phosphorylation of Akt and mTOR were reduced (P<0.01). CONCLUSIONS:Rapamycin could inhibit invasion and metastasis of HeLa cell via Akt/mTOR signal pathway.

Objective To establish the fingerprints analysis of the methanol extracts of nutmeg,and study quality uniformity of nutmeg in different areas.Methods A ZORBAX EclipseXDB-C18 (4.6 mm?150 mm,5 ?m) column was used.The mobile phase consisted of methanol-acetonitrile-water (25∶35∶40),the flow rate was 1 mL/min,the column temperature was 30 ℃,the detective wavelength was 270 nm.Dehydrodiisoeugenol was used as reference compound.Results Fingerprint of nutmeg was established,consisted of l7 common peaks.The similarity of fingerprints was over 0.9.Conclusion The fingerprints of nutmeg in different areas have no differences.This method is accurate,reliable and provides a scientific basis for the quality control of nutmeg.

BACKGROUND:Compared with the conventional composite resin, the 3M Z350 nano-resin has good wear
resistance, physical mechanical properties, and polishing, and exerts a lower irritation to the dental pulp. Besides fil ing materials, a reliable tooth-prosthesis bonding interface is necessary for resin bonded repairs.
OBJECTIVE:To compare the clinical effects of self-etch bonding Adper Easy One and total-etch bonding Single Bond 2 on nano-resin bonding restoration of the anterior teeth.
METHODS:120 anterior teeth with vital pulp, which had defects at the incisal ends and were to be restored with nano resins, were divided into two groups randomly. Two kinds of adhesives, self-etch adhesive and total-etch adhesive, combined with nano-resin were used to restore the teeth. The patients were re-examined immediately, 6 months, 1 year and 2 years after the treatment. The fil ings, teeth and pulps of patients were examined,
including whether the prosthesis and tooth color were coordinated, whether the gap between the prosthesis and the teeth were sealed, whether the surface of the prosthesis was intact with no loose, whether the prosthesis and teeth had no staining and secondary caries, whether the condition of the tooth pulp had hot or cold
stimulation-induced pain.
RESULTS AND CONCLUSION:No significant difference in the fil ing effects was found between the two groups when the patients were re-examined immediately, 6 months and 1 year after the treatment (P>0.05). The pulp
lesions of the self-etching group were fewer than those of the total-etch group 2 years after the treatment (P<0.05). Self-etching group had 1, 6, 0, 2 cases and total-etch group had 0, 2, 1, 2 cases in uncoordinated color, edge seal,
incomplete restoration and secondary caries, respectively. No statistical y significant differences were found in these four aspects between the two groups (P>0.05). The 2-year fol ow up showed a low incidence of pulp lesions and satisfactory clinical performance after 3M Z350 nano-resin working with self-etching bonding system in the nano-resin fil ing of
anterior teeth with vital pulp.

Objective To investigate the value of CysC-based CFR in comparison with creatinine-based GFR (CG-GFR and MDRD-GFR) as an accurate serum marker in the prediction of early diabetic nephropathy. Methods A tatal of 133 type 2 diabetic patients (74 males and 59 females, aged 58.1 ±12.3) were enrolled. The level of diabetic nephropathy (normoalbuminuric, microalbuminuric and macroalbuminuric) was staged and estimated GFR based on serum creatinine and cystatin C(CysC) was calculated. The plasma clearance of Tc-DTPA, serum CysC, creatinine, BMI, HbA1c, serum lipid and blood pressure were measured. Results 99mTc-DTPA-GFR was used as golden standard. At 90 and 75 ml· min-1· (1.73 m2)-1 cut-points, diagnostic efficiencies of CysC-GFR (89% and 92%) were better than those of CG-GFR (79%=86%, P=0.004, 0.04) and MDRD-GFR (80%-86%, P=0.02, 0.04). At 60 ml · min-1 · (1.73 m2)-1 cut-point, diagnostic efficiencies of CysC-GFR,CG-GFR and MDRD-GFR were 92%, 90% and 92% respectively (P= 0.49, P=0.71). The Logistic regression analyses showed that retinopathy, HbA1c, CysC, diabetic duration, and CysC-GFR were indicators to predict the development of microalbuminuria. Conclusion CysC-GFR is more valuable than CG-GFR and MDRD-GFR in the prediction of early diabetic nephropathy and should be applied clinically.

ObjectiveTo observe β cell function in newly diagnosed diabetic patients with ketosis before and in remission period and evaluate its classification and predictive value.MethodsA total of 206 patients newly diagnosed as diabetic ketosis who had been treated with intensive insulin therapy in our hospital and entered in the honeymoon after the withdraw of insulin therapy were followed for 36 months from onset of diabetes.They were divided into two groups of type 1 and type2 diabetes ( group A and B),according to the dependence or independence on insulin treatment. The β cell function of the two groups before and in remission period was compared by oral glucose tolerance test (OGTF).β cell function was measured with the AUC of insulin and C-peptide and homeostatic model assessment β-cell function (HOMA-β),while homeostatic model assessment insulin resistant (HOMA-IR) for insulin resistant.The duration of the honeymoon and the change of insulin and C-peptide curve before and in honeymoon were also observed.ResultsThe AUC of insulin and C-peptide,the HOMA-β and the HOMA-IR before and after the intensive insulin treatment were lower in group A than that in group B [ before the insulin treatment:(10.18 ±2.36)mIU · h · L-1 vs (20.28 ±6.89)mIU · h · L-1,(1.56 ±0.53) μg · h · L-1 vs (3.75 ±0.67) μg · h · L-1,3.68 ± 1.08 vs 18.20 ±6.59,1.22 ±0.49 vs 3.06 ± 1.54,respectively;after the insulin treatment:(29.86 ± 8.65 ) mIU · h · L-1 vs (93.35 ± 19.42 ) mIU · h · L-1,( 3.99 ± 0.79 )μg · h · L-1 vs ( 12.54 ±3.83) μg · h · L-1,8.50 ±2.46 vs 56.17 ± 19.42,0.63 ±0.56 vs 1.42 ±0.78,respectively ].The duration of the honeymoon in group A was significantly shorter than in group B [ (7.9 ±5.2) months vs (20.9 ± 9.9 ) months ].In oral glucose insulin and C-peptide release test,the peak of insulin and C-peptide releasing curve in group A was brought forward by a half to 1 hour after intensive treatment while delayed in group B by 1 or 2 hours.The releasing peak of insulin and C-peptide in group A was less than two folds of the basic value,while four to ten fold of the basic value in group B.The positive ratio of glutamic acid decarboxylase antibody,insulin autoantibody and insular cellular antibody in group A and group B were 21.2％ vs 4.8％,18.1％ vs 3.3％,9.2％ vs 10.6％,respectively.ConclusionsOf all the patients newly diagnosed as diabetes ketosis who had entered into the honeymoon after intensive insulin therapy,91％ were type 2 diabetes.Inferior β cell function before insulin therapy,weaker remission after insulin therapy and shorter duration of remission period suggest the classification of type 1 diabetes.

Objective:To study the influence of vitamin C ( VC) on dendritic cells ( DC) ,and detect DC maturation,to provide a feasible method and thought for quickly preparating DC vaccines.Methods:Collected the peripheral blood (about 50 ml) from healthy volunteers,and isolated peripheral blood mononuclear cells with lymphocyte separation medium and obtain DC.With stimulating with different concentrations of VC for (24 h),then washed with PBS,and set up blank control group (V0).The expression of DC surface co-stimulating molecules CD80/86 and HLA-DR, CD40 was detected by flow cytometry.By setting the concentration gradient and time gradient, exciting optimal concentration and stimulating time of VC on DC, and analyzed the reasons of VC promoting DC maturation.Results:VC could effectively stimulate DC,CD80/86 expression had significantly increased contrast to the blank control group (V0).And the experiments show that VC’s best stimulating concentration was 1 mmol/L,and on the third day,the CD80/86 expression rate of VC group was (78.6±4.6) %,and blank control group V0 was (34.1±5.7) %.DC surface HLA-DR expression:VC (56.8± 4.4) %,blank control group V0 (25.4 ±4.7) %,the difference between two groups was statistically significant,P<0.05.CD40 and CD40L expression and results show that VC 2.5 mmol/L group of CD40 expression rate up to (59.3±3.7) %,while V0 group was only (11.1 ±2.4) %,that illustrate VC could significantly regulate CD40 expression on DC surface,but CD40L not reflect.Fluorescence mi-croscope results showed that DC’ s antigen catching ability was also significantly promoted.Conclusion:VC can significantly regulate DC maturity,and may up regulate CD40,thus promoting the express of CD80/86 and HLA-DR.When the concentration is 1 mmol/L,VC expresses the strongest regulation function on DC.

Objectives Understanding the characteristic changes of Keshan disease (KD) in different epidemic period to provide reference basis for prevention and teatment. Methods On the basis of medical record as fundamental element, the relative conditions of Keshan disease's prevailing and spreading period were compared. Results In high incidence years, familial aggregation [accounted for 12%(6/50)] and seasonal aggregation were found, and KD cases occurred mainly from May to September, which was 78% (39/50)of the total cases in the whole year. Circulatory dysfunction(gallop rhythm, pulmonary role, jugular venous engorgement, cyanosis of lips) was more severe in high incidence years than that in low incidence years(X2=8.53,P<0.01). The average age of incidence was (4.07±1.46) years old in high incidence years and (6.11±2.71) years old in low incidence years. The type constitution in high incidence years was significantly different from that in low incidence years (X2=40.68, P<0.01), and chronic type of KD accounted for 22.85%(707/3094),46.09%(53/115), respectively, in high and low incidence years. Conclusions Making a further research of seizure of disease, and improving diagnosis and cure management level are also the important content for prevention and cure research work of Keshan disease at right time.

OBJECTIVE:To establish the HPLC fingerprints for Herba clematidis in northeast. METHODS:HPLC was per-formed on the column of Hedera ODS-2 C18 with mobile phase of acetonitrile-0.5% phosphoric acid solution(gradient elution)at a flow rate of 1.0 mL/min,detection wavelength was 338 nm,column temperature was 30 ℃,and the injection volume was 20 μL. Using rutin as as a reference,the HPLC profiles of 10 batches of H. clematidis were determined,Similarity Evaluation Software for Chromatographic Fingerprint of Traditional Chinese Medicine(2004A edition) was used for the common peaks identification and similarity evaluation. RESULTS:There were 16 common peaks in the 10 batches of H. clematidis,similarity degree was higher than 0.9. It was proved that the HPLC profiles and control fingerprint profile of 10 batches of H. clematidis had good consistency. CONCLUSIONS:The established fingerprints can provide reference for the identification and quality evaluation of H. clematidis in northeast.