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.

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.

Neurolymphomatosis(NL) is characterized by lymphomatous infiltration of the peripheral nervous system. We report a case of neurolymphomatosis(NL) which was confirmed by sural nerve biopsy. Sural nerve specimen from a 49-year-old female patient with weakness of limbs was examined with routine histochemical and immunohistochemistry staining, in which the first antibodies against CD3, CD20, CD45, CD45RO and CD68 were used. Numerous T-lymphoma cells invaded in the adipose tissue of epineurium of sural nerve. The nerve biopsy showed marked axonal degeneration of myelinated fibers. The clinical and histopathologic findings confirmed the diagnosis of neurolymphomatosis.

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.

Objective To investigate the effects of hydrogen rich water on the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and oxidative stress in rats with traumatic brain injury (TBI).Methods Ninety healthy male Sprague-Dawley (SD) rats were randomly divided into sham operation group, TBI group and hydrogen rich water treatment group (HW group), with 30 rats in each group.TBI model was reproduced by the modified Feeney weight dropping method.The skulls of rats in sham operation group underwent only craniotomy without direct hit.The rats in HW group received brain injury by hitting after craniotomy, followed by injection of hydrogen rich water (5 mL/kg) intraperitoneally once a day for 5 days after successful reproduction of the model.The rats in sham operation group and TBI group were given an equal amount of normal saline in same manner.Six rats from each group were sacrificed at 6, 12, 24, 48 hours and 5 days after evaluating neurological severity scores (NSS).The brain tissue in injured ipsilateral cortex was harvested.The activity of catalase (CAT), glutathione peroxidase (GSH-Px), and content of malondialdehyde (MDA) were determined by spectrophotometry.The expressions of mRNA and nucleoprotein of Nrf2 were determined by quantitative real-time polymerase chain reaction (RT-qPCR) and Western Blot.The pathological changes were observed with microscopy after hematoxylin and eosin (HE) staining.Results ① NSS score:compared with TBI group, NSS in HW group at 12, 24, 48 hours and 5 days were significantly decreased (12 hours: 9.83±2.32 vs.13.17±2.71, 24 hours: 9.83±2.79 vs.13.50±2.43, 48 hours: 7.50±2.07 vs.11.83±2.14, 5 days:5.50 ± 1.87 vs.10.50 ± 2.43, all P ＜ 0.05).② Compared with sham operation group, the activity of GSH-Px and CAT in TBI group were markedly declined after operation, while the MDA content was elevated significantly, especially at 24 hours [CAT (kU/g): 1.080±0.312 vs.3.571 ±0.758, GSH-Px (kU/g): 9.195±3.173 vs.32.385± 10.619, MDA (μmol/g): 12.282±2.896 vs.4.349± 1.511, all P ＜ 0.01].Compared with TBI group, the parameters in HW group were improved, and they were similar as sham operation group.③ RT-qPCR: no significant difference was found in the expression of Nrf2 mRNA at each time point in three groups.④ Western Blot: the expression of Nrf2 nucleoprotein (gray value) in TBI group was apparently higher than that in sham operation group, and peaked at 24 hours (0.703 ± 0.262 vs.0.238 ± 0.120, P ＜ 0.05), and the expression in HW group was obviously higher than that in TBI group, especially at 24 hours (1.110 ± 0.372 vs.0.703 ± 0.262, P ＜ 0.05).⑤ HE staining: the brain structure in sham operation group was found to be intact.However, there were different degrees of pathological changes at each time in TBI group, especially at 24 hours.The pathological damage of brain tissue in HW group was significantly milder.Conclusions Hydrogen rich water can up-regulate the expression of Nrf2, and reduce oxidative damage of traumatic brain injury in rats.Nrf2 can up-regulate the expression of its downstream antioxidant enzymes, which may be the mechanism of the upregulation expression of Nrf2 in the study.

Objective To investigate the role of NMDA receptors in central medial thalamus (CMT) in the unconscious?ness induced by general anesthesia. Methods A total of 60 rat models for microinfusion were assigned into 4 groups (n=15 for each group). After induction with propofol, 10 mmol/L (NMDA10 group), 20 mmol/L (NMDA 20 group) and 40 mmol/L (NMDA40 group) of NMDA and normal saline (group C) with equal volume were microinfused into CMT. The incidence of purposeful movement and recovery time of righting reflex were observed in each group respectively. Infusion sites were local?ized by histological method. Results When the microinfusion site localized within CMT, comparing with group C, the recov?ery time of righting reflex reduced notably in three NMDA groups (P<0.05). The recovery time was significantly shorter in NMDA20 group and NMDA40 group than that of NMDA10 group. The incidence of purposeful movement during propofol an?esthesia was higher in NMDA20 group and NMDA40 group than that of group C (P<0.05). When the microinfusion site lo?calized out of CMT, the recovery time of righting reflex was remarkably longer than that within CMT in three NMDA groups (P<0.05), and there was no significant difference in the incidence of purposeful movement and recovery time between four group (P>0.05). Conclusion Microinfusion of NMDA agonist into CMT reverses propofol anesthesia, indicating that NMDA receptor in CMT may contribute to the propofol-induced unconsciousness.