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 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.

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.

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 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.

Objective To investigate the effects of pioglitazone on angiotensin Ⅱ(AngⅡ) -induced proliferation and collagen typeⅠ expression of adventitial fibroblasts (AF). Methods The AFs were isolated from rat thoracic aorta. MTT colorimetry and flow cytometry were used to study the effects of pioglitazone on proliferation and cell cycles of AF. The expression of collagen typeⅠ regulated by pioglitazone was examined by the method of Western blot. Results Pioglitazone inhibited the proliferation of AF in a dose-dependent manner, and the most marked effect could be observed at the concentration of 10?10-6 mol/L for pioglitazone (P

Objective To investigate the effect of hydrogen-rich water on cerebral edema and aquaporin 1 (AQP1) expression in rats with traumatic brain injury (TBI).Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into sham operation group,TBI model group,hydrogen-rich water treatment group (H group),with 30 rats in each group.TBI model was reproduced by weight dropping method.The skulls of rats in sham operation group underwent only craniotomy without direct hit and with bone wax sealed suture.5 mL/kg of hydrogen-rich water injection was given intraperitoneally after model reproduction in H group,and equal amount of normal saline was given in sham and TBI groups,once a day for both groups for 5 days.Six rats from each group were sacrificed at 6,12,24,48 hours and 5 days after evaluating neurological severity scores (NSS).The cerebral cortex was harvested,and the pathological changes in morphology of brain tissue were observed with light microscope.The positive expression of AQP1 in cerebral cortex was observed with immunohistochemistry by light microscopy,the AQP1 mRNA expression in cerebral cortex was determined by real-time fluorescent quantization reverse transcription-polymerase chain reaction (RT-PCR),and the AQP1 protein expression in cerebral cortex was determined by Western Blot.Results ① All rats in sham operation group had a NSS of zero at each time point.NSS of TBI group was obviously raised with time prolongation,and peaked at 24 hours followed by a lower tendency,while the score in H group was significantly lower than that of TBI group,and the difference was the most obvious at 24 hours as compared with TBI group (9.83 ± 2.78 vs.13.50± 2.42,P ＜ 0.05).② It was shown by light microscope that in the TBI group there were pathological changes in cerebral cortex,including obvious irregular arrangement of nerve cells,cerebral edema,obvious bleeding,especially at 24 hours,then the cerebral edema became vanished gradually;and the positive expression of AQP1 in the pia mater at all the time points in the TBI group was significantly increased,and it was most obvious at 24 hours.Compared with TBI group,the pathological changes at time points of 12 hours to 5 days in H group was significantly lessened,and the positive expression of AQP1 in the cerebral pia mater was reduced obviously.③ Compared with sham operation group,the mRNA and protein expressions of AQP1 in cerebral cortex in TBI group were significantly elevated,peaked at 24 hours [AQP1 mRNA (2-△△Ct):7.50±0.26 vs.1,AQP1 protein (gray value):1.986±0.110 vs.0.336±0.034,both P ＜ 0.05],then they gradually declined.The mRNA and protein expressions of AQP1 in cerebral cortex were significantly decreased after hydrogen-rich water treatment [24-hour AQP1 mRNA (2-△△Ct):5.40±0.21 vs.7.50±0.26,24-hour AQP1 protein (gray value):1.246±0.137 vs.1.986±0.110,both P ＜ 0.05].Conclusions The up-regulation of AQP1 mRNA and protein in ratst cerebral cortex after TBI perhaps participates in edema formation which might be involved in the pathophysiology of cerebral edema in TBI.Early treatment with an intraperitoneally injection of hydrogen-rich water is capable of attenuating the extent of TBI-induced up-regulation of AQP1 mRNA and protein,alleviating cerebral edema,and achieving its protective effects.

Objective: To assess the values of conventional vascular ultrasound (US) and superb micro vascular imaging (SMI) for diagnosing carotid artery stenosis in relevant patients. Methods: A total of 37 patients of extra cranial carotid stenosis (with 70 blood vessels) treated in our hospital from 2014-08 to 2015-03 were retrospectively studied. Digital subtraction angiography (DAS) examination was used as golden standard, the diagnostic efifcacies for carotid artery stenosis by US and by SMI were compared. Results: The accuracy, sensitivity and specificity for diagnosing carotid stenosis by US were 81.42%, 83.33% and 80.95%; by SMI were 91.43%, 92.16% and 89.47% respectively. Conclusion: US and SMI showed good agreement for diagnosing carotid artery stenosis, while SMI was superior to US for accurately assess the degree of carotid stenosis, it might be used as a more reliable method for evaluating carotid plaque and stenosis in relevant patients.

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.