Objective: To investigate the bioactive constituents of coral Scleronephthya sp. collected from the South China Sea. Methods: The compounds were isolated and purified using repeated column chromatographies on Sephadex LH-20, normal- and reversed-phase silica gels, and RP-HPLC. The structures of the compounds were elucidated based on the detailed spectroscopic analysis in combination with reported data. The in vitro antimicrobial activity of these compounds were assessed by an agar diffusion test. Results and conclusion: Three 5α, 8α-epidioxysterols, two alkyl glycerol ethers and 1, 2-O-alkyl glycerol were isolated from Scleronephthya sp. ; these compounds displayed different levels of antifungal and antibacterial activities in bioassay in vitro. Compound 5 inhibited the growth of Gram-negative bacterium Escherichia coli and the Gram-positive bacterium Bacillus megaterium. Compound 6 showed marked antifungal activity against Microbotryum violaceum. Compounds 1-4 demonstrated a weak antimicrobial activity.

OBJECTIVE:To observe clinical efficacy and safety of Ebastine tablet combined with Chushi zhiyang ointment in the treatment of hand keratinizing chapped eczema. METHODS:135 cases of hand keratinizing chapped eczema were divided into control group A(45 cases),control group B(43 cases)and treatment group(47 cases)according to treatment regimen. Control group A was orally given Ebastine tablet,10 mg each time,qd;control group B was given Chushi zhiyang ointment alone,twice a day,morning and evening,applying thin layer of ointment on the affected area;treatment group was given same dose of Ebastine tablet orally and applied Chushi zhiyang ointment on the affected area. 3 groups received treatment for consecutive 4 weeks. Clinical efficacies of 3 groups were observed as well as the scores of pruritus,skin lesion area,keratinization,rhagades and VAS before and after treatment. The occurrence of ADR was compared among 3 groups. RESULTS:The total effective rate of treatment group was 68.09%,which was significantly higher than that of group A(42.22%)and control group B(16.28%),with statistical significance(P<0.05). There was no statistical significance in the scores of pruritus,skin lesion area,keratinization,rhagades and VAS among 3 groups before treatment(P>0.05). After treatment,above scores of 3 groups decreased significantly,and those of treatment group were significant-ly lower than those of control group A and B,with statistical significance(P<0.05). There was no statistical significance in the inci-dence of ADR among 3 groups(P>0.05). CONCLUSIONS:Ebastine tablet combined with Chushi zhiyang ointment is effective for hand keratinizing chapped eczema,and can significantly improve the skin of patients with good safety.

AIM:To evaluate the inhibitory effect of FRNK on the phosphorylation of FAK and apoptosis in hepatic stellate cells (HSCs). METHODS: After stimulated with fibronectin, HSCs was transfected with FRNK plasmid by cationic liposome method. The apoptosis of FRNK-induced HSCs was examined by Annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscope. The protein levels of FRNK, FAK and p-FAK (Tyr397) in HSCs were assayed by Western blotting, and RT-PCR was used to detect the expression of mRNA. RESULTS: The expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSCs in vitro. The apoptotic rate in HSCs exposed to FRNK plasmid for 48 h was higher than that in the non-FRNK plasmid group [(25.37?1.92) % vs (9.28?1.05) %, P

Objective To investigate the effects of human papilloma virus 16 E6 (HPV16 E6) on endoplasmic reticulum (ER) stress-autophagic response in the cervical cancer C33A cells.Methods Polymerase chain reaction was used for detecting the integration of HPV DNA.The eukaryotic expression vector of HPV16 E6 was constructed and transfected via lipofectamine into C33A cells.Experimental cells were classified into 3 groups:pcDNA3.1--HPV16 E6 group,pcDNA 3.1-group and C33A group.Western blot was used to measure expression of protein of HPV16 E6,Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 in transfected cells.Results here was no HPV DNA integration in C33A cells that were confirmed as the intervention cells.Eukaryotic expression vector pcDNA3.1--HPV16 E6 was constructed successfully.The eukaryotic expression vector pcDNA3.1--HPV16 E6 significantly improved the expression of protein of HPV 16 E6 in C33A cells.The protein expression of Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 were significantly improved after transfection with vector pcDNA3.1 +-HPV16 E6 (P ＜ 0.05).Furthermore,LC3 Ⅱ protein level was reduced by treatment with ER stress inhibitor.Conclusion HPV16 E6 can improve autophagy through the ER stress pathway,and this response may play an important role in the process of HPV16 E6 inducing cervical cancer,providing one of the new strategies for gene therapy of cervical carcinoma.

AIM: To investigate the effect of FAK - related non - kinase ( FRNK) on the expression of membrane - type matrix metalloproteinase -1 ( MT1 - MMP) in hepatic stellate cells ( HSC). METHODS:FRNK were trans-fected into HSCs by cationic liposome method. The protein levels of FRNK in HSC were assayed by Western blotting. The levels of MT1 - MMP were determined by RT - PCR for mRNA and by Western blotting for protein, respectively. RESULTS: The up -regulated expression of FRNK protein was observed and it was at 48 h after transfection that the FRNK protein content was the highest ( P < 0.05 ). The expressions of MT1 - MMP mRNA and protein were also up - regulated by the transfection of FRNK, and it was at 48 h after transfection that the MT1 - MMP protein content was significantly increased. CONCLUSION: The mRNA and protein of FRNK were over - expressed in HSC transfected with the gene of FRNK. The inhibitory effect of FRNK on the collagen synthesis in HSC may be through the up - regulation of MT1 - MMP.

ObjectiveTo investigate the effect of isoflurane preconditioning on glutamate-induced apoptosis in rat neuronal PC12 cells.MethodsThe PC12 cells were cultured for 5 d with nerve growth factor in vitro.The cells were seeded into 6-cm-diameter culture dishes (3 ml/dish) or 6-well plates (2 ml/well) with the density of 5 × 104/ml and randomly divided into 4 groups (n =18 each): normal control group (group C); glutamate group (group G) ;glutamate + isoflurane group (group GI) and glutamate + isoflurane + xestospongin C (an antngon of inositol trisphosphate receptors) group (group GIX).The neuronal PC12 cells were exposed to glutamate 500 μmol/L in groups G,GI and GIX.The neuronal PC12 cells were exposed to 1.2％ isoflurane for 2 h in groups GI and GIX.Xestospongin C was added to the culture medium immediately before isoflurane preconditioning.Glutamate was added to the culture medium at 10 min after isoflurane preconditioning in groups GI and GIX.The cells were collected from six dishes or wells in each group after being incubated with glutamate for 20 min.The apoptosis and mitochondiral membrane potential (MMP) were assessed by flow cytometry.Intracellular Ca2+ concentration ([ Ca2+ ] i)was detected by confocal fluorescence microscopy.ResultsCompared with group C,the apoptotic rate and [Ca2+ ]i were significantly increased and MMP was decreased in groups G and GIX ( P ＜ 0.01 ),but there was no significant difference in the variables mentioned above in group GI (P ＞ 0.05).Compared with group G,the apoptotic rate and [ Ca2 + ]i were significantly decreased and MMP was increased in groups GI and GIX ( P ＜ 0.05 or 0.01).Compared with group GI,the apoptotic rate and [Ca2+ ]i were significantly increased and MMP was decreased in group GIX ( P ＜ 0.01 ).ConclusionIsollurane preconditioning can inhibit apoptosis in rat neuronal PC12 cells by activating inositol trisphosphate receptors,inhibiting Ca2+ release from the endoplasmic reticulum and increasing MMP.

Objective To compare the MRI manifestations and characteristics of ≤ 3 cm nonepithelial hepatic angiomyolipomas (HAML)and small hepatocellular carcinoma (SHCC),then improve the preoperative diagnostic accuracy.Methods A retrospective analysis of 20 patients ≤3 cm nonepithelial HAML and 26 cases of SHCC,confirmed by clinical pathology,with both in clinical data and MRI characteristics.Results ≤3 cm nonepithelial HAML commonly occurs in women;The enhancement patterns“wash in and wash out”in 1 1 cases,6 cases “wash in but slow out”,3 cases with delayed enhancement;Mature adipose tissue found in 5 cases, 3 cases has pseudocapsule enhancemen in delayed phase,13 cases can see central vessels;While SHCC often occurs in men,with“wash in and wash out”enhancement pattern in 23 cases,1 7 cases with pseudocapsule enhancemen in delayed phase,12 cases can see central vessels in lesions.ADC values for SHCC was significantly lower than that for ≤3 cm nonepithelial HAML,the ADC values of SHCC significantly lower than the surrounding liver parenchyma;The ADC values between ≤3 cm nonepithelial and liver parenchyma around has no significant differences.ADC values of liver parenchyma with liver cirrhosis was lower than that without cirrhosis.Conclusion The existence of mature adipose tissue,the MRI enhancement pattern and the value of ADC can help to distinguish between ≤3 cm nonepithelial HAML and SHCC,then improve the preoperative diagnostic accuracy.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Clear Cell ; metabolism ; pathology ; Adult ; CA-125 Antigen ; metabolism ; Carcinoembryonic Antigen ; metabolism ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; surgery ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; Ki-67 Antigen ; metabolism ; Lymph Node Excision ; Membrane Proteins ; metabolism ; Neoplasm Invasiveness ; Uterine Cervical Neoplasms ; metabolism ; pathology ; surgery

Aim To investigate whether human umbili-cal cord mesenchymal stem cells(hUC-MSCs)exposed to inflammatory conditions could release large amounts of exosomes to induce regulatory T cells(Treg).Meth-ods hUC-MSCs were isolated by enzyme digestion method.(In vitro)interferonγ(IFN-γ)was added in-to hUC-MSCs to mimic inflammatory microenviron-ments,then exosomes were extracted from the superna-tant of normal conditional medium or IFN-γpretreated hUC-MSCs.Both sources of exosomes,Nor-hUC-exo and IFN-γ-stimulated hUC-exo, were identified by Nanoparticle Trafficking Analysis (NTA )and Western blot for the exosome-enriched protein CD63 .Next,hu-man peripheral blood mononuclear cells (PBMCs ) stimulated with PHA were respectively co-cultured with hUC-MSCs,IFN-γ-pretreated-hUC-MSCs,hUC-MSCs exosomes or IFN-γ-stimulated-hUC-MSCs exosomes for 5 days to assess the exosomes-T cells communication. The proliferation rate of PBMCs and frequency of CD4 +/CD25 +/Foxp3 + Treg were measured by flow cytometry.Results The isolated cells from human um-bilical cord tissue,which were positive for CD73, CD44,CD29,CD90 and HLA-ABC,but were nega-tive for CD31 and CD34,were mesenchymal stem cells indeed.After IFN-γtreatment,hUC-MSCs secreted nu-merous exosomes(P<0.05 ).Morerover,there was a significantly higher level of CD63 ,but no difference in diameter between Nor-hUC-exo and IFN-γ-stimulated hUC-exo.IFN-γ-stimulated hUC-exo had a superior a-bility compared with Nor-hUC-exo to suppress the pro-liferation of PHA stimulated PBMCs due to their upreg-ulation of the percentage of Treg (1 1.53 ±0.88% vs 6.60 ±0.56%,P <0.01 ).Conclusion hUC-MSCs could promote the expression of Treg to modulate im-munosuppression through exosomes,especially for IFN-γ-licenced exosomes,which might carry much immu-notherapeutic potential.

Objective To evaluate the detection of membrane neutrophilic alkaline phosphatase ( mNAP) by flow cytometry in diagnosis of bloodstream infection .Methods A total of 298 patients with suspected bloodstream infections admitted in the First People ’ s Hospital of Lianyungang during June 2013 and October 2014 were enrolled;80 healthy subjects in physical examination center were also enrolled as the control group.Bloodstream infection was diagnosed by blood culture and mNAP was detected by flow cytometry.Serum levels of procalcitonin (PCT) and C-reactive protein (CRP) were detected by electro-chemiluminescence (ECL) and immune scatter turbidimetry , respectively.The value of mNAP, PCT and CRP in diagnosing bloodstream infection was determined by receiver operating characteristic ( ROC) curve. Results Among 298 patients, 109 were confirmed with bloodstream infections , including 43 patients with Gram-positive bacterial infections and 66 with Gram-negative bacterial infections .The median levels of CRP , PCT and mNAP in bloodstream infection group were 138.71 mg/L, 7.04 ng/mL and 13 929 AB/c, which were significantly higher than those in healthy control group (1.50 mg/L, 0.12 ng/mL, 1 831 AB/c;U=5.00, 48.50 and 65.01, P<0.01).The expression of mNAP in Gram-positive bacterial infection group was 9 598 ( 6 064-11 643 ) AB/c, which was significantly lower than that in Gram-negative bacterial infection group [16 512 (11 654-22 001) AB/c] (U=250.00, P<0.01).ROC curve analysis showed that, the areas under the curve (AUCs) of mNAP, PCT and CRP in diagnosing bloodstream infection were 0.987, 0.962 and 0.901.When 4 578AB/c, 0.90 ng/mL and 13.50mg/L were taken as optimal cut-off values, the sensitivities of mNAP, PCT and CRP in diagnosis of bloodstream infection were 95.8%, 93.0%and 90.3%; the specificities were 97.8%, 95.6% and 85.5%, respectively.Conclusion Among mNAP, PCT and CRP, mNAP is of the highest value in diagnosing bloodstream infection , and may be used as a biomarker for clinical diagnosis of bloodstream infection .