[Objective] To investigate the value of HBV-M and HBV DNA of newborns born to HBsAg-positive mother, which were tested before combined immunization of hepatitis B. [Method] A total of 420 infants born to HBsAg-positive mothers delivered in Obstetric Department of the Third Affiliated Hospital of Sun Yat-Sen University from June 2006 to February 2008 were followed up at least 6 months and rechecked HBV-M to confirm the diagnosis of HBV intrauterine infection, which included 33 HBsAg or HBV DNA positive newborn babies and 6 newborns with both HBsAg seropositive and HBV DNA seropositive. [Result] HBV intrauterine infection rate was 0.95%. Using newborn both HBsAg positive and HBV DNA positive as diagnostic criterion to diagnose HBV intrauterine infection, the positive likelihood ratio was 208.3, while using newborn HBsAg positive or HBV DNA positive as diagnostic criterion, it was 14.3. [Conclusion] Newborn both HBsAg positive and HBV DNA positive obtained before combined immunization of hepatitis B may predict HBV intrauterine infection, and it may play as a clinical index of preliminary diagnosis of HBV intrauterine infection.

OBJECTIVE:To optimize the stir-bake with saltwater processing technology for Fujian Alismatis rhizoma,and es-tablish its HPLC fingerprint. METHODS:Using 23-acetyl alisol B,total triterpenoids contents and appearance as comprehensive in-dexes,single factor experiment and orthogonal test were employed,3 factors'levels including the quantity of salt,processing tem-perature and time were optimized. HPLC was used to develop fingerprints of 10 batches of Fujian Alismatis rhizoma at wavelength of 208,245 nm;the similarity between fingerprints and control profile was compared by using software. RESULTS:The optimal technology was as follow as 2 kg salt dissolved with 10 kg water for each 100 kg Fujian Alismatis rhizoma,moistening for 1 h, stir-frying for 8 minutes under 100 ℃. In verification test,the appearance of 3 batches of processed products were all in line with requirements,average comprehensive score was 93.94 (RSD=6.63%,n=3);23-acetyl alisol B and total triterpenoids contents were stable(RSD=7.41%,7.39%,n=3),respectively. Totally 17 and 10 common peaks were marked in 208 nm and 245 nm re-spectively;similarities of 10 batches of samples'fingerprints were higher than 0.9. CONCLUSIONS:Optimized stir-bake with salt-water technology is reasonable,feasible,and reproducible;the stability of common peaks of established fingerprints can conduct ef-fective quality evaluation for Fujian Alismatis rhizoma processed by saltwater.

Objective To analyze the role of a key intracellular signaling molecule GSK3β in hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS).Methods Using mouse hepatoma cell lines(Hepa 1) as cell apoptosis model triggered by tunicamycin,an endoplasmic reticulum stress inducer.One hour before Hepa 1 apoptosis induced by tunicamycin,SB216763 specifically inhibited the activity of GSK3β.Living cells/apoptotic cells were detected using acetoxymethyl (AM)/propidium iodide (PI) staining; Furthermore,the measurement of lactate dehydrogenase(LDH) of cell culture supernatant to evaluate the apoptosis.We detect p-GSK3β,GSK3β,the ERS-related protein(GRP78,CHOP and caspase-12) and caspase-3,cleaved caspase-3 protein expression using Western blot.Results Endoplasmic reticulum stress induced by tunicamycin promotes GSK3β activity; Inhibition of GSK3β activity alleviates endoplasmic reticulum stress:the expression of GRP78,CHOP and caspase-12 expression are inhibited.At the same time,GSK3β activity inhibition significantly reduced the endoplasmic reticulum stress-induced apoptosis:compared to cell apoptosis model group,the intervention group of SB216763 showed that the level of LDH decreased significantly,and PI staining of apoptotic cells was also significant reduction.Western blot results showed that the inhibition of GSK 3 β activity reduced reactive cleaved caspase-3 protein.Conclusion GSK3β is an important signaling molecule in the apoptosis pathway induced by endoplasmic reticulum stress ;Endoplasmic reticulum stress promotes hepatocyte apoptosis by mediating GSK3β.

AIM: To explore the roles of heat shock protein 90 (HSP90) in the blockage of hydrogen sulfide (H2S) against chemical hypoxia-mimetic agent (cobalt chloride, CoCl_2)-induced oxidative stress injuries in H9c2 cardiac cell. METHODS: H9c2 cells were treated with CoCl_2 to set up the chemical hypoxia-induced the model of cardiomyocyte injury. Sodium hydrosulfide (NaHS, a H2S donor) was added into medium for 30 min before CoCl_2 treatment. ATP content was detected by high performance liquid chromatogram (HPLC). Mitochondrial membrane potential (MMP) was measured by rhodamine123 (Rh123) staining and photofluorography. The activity of superoxide dismutase (SOD) was observed using a SOD kit. The expression of heme oxygenase-1 (HO-1) was evaluated by Western blotting. RESULTS: CoCl_2 at concentration of 600 μmol/L significantly reduced SOD activity, ATP level and MMP, and enhanced the expression of HO-1 in H9c2 cells. Pretreatment with 400 μmol/L NaHS dramatically inhibited the cytotoxicity induced by CoCl_2, increased SOD activity, ATP level and MMP, decreased HO-1 expression. 17-allylamino-17 demethoxygeldanamycine(17AAG), an inhibitor of HSP90, obviously blocked the inhibitory effect of H2S on the CoCl_2-induced cytotoxicity, reduced the levels of ATP and MMP, increased HO-1 expression. However, no significantly influence on SOD activity was observed. CONCLUSION: HSP90 may mediate the cardioprotection of H2S via inhibiting the oxidative stress induced by chemical hypoxia.

Objective To investigate the dynamic expression of the drug resistance protein P-glycoprotein (P-gp) within 72 hours in the pentylenetetrazol (PTZ)-induced status epilepticus (SE) model, and to identify the optimal detection time to inhibit P-gp. Methods mRNA and protein expressions of P-gp in rats hippocampal tissue were detected by using immunohistochemistry , RT-qPCR and Western blot at different time points after modeling (0, 3, 6, 12, 24, 48, 72 h). Results The mean density of P-gp protein in the hippocampus of status epilepticus model was 0.325 1 ± 0.008 2 at 24 h, and was 0.396 3 ± 0.016 8 at 48 h, which were consistently higher than those of the control group (P < 0.05, P < 0.01, respectively). Results of qRT-PCR showed that MDR1a expression was significantly upregulated at 24 h and at 48 h (P < 0.05, P < 0.01, respectively). Western blot assay revealed that P-gp protein was also significantly increased at 48 h after seizures (P < 0.05). Conclusions The upregulation of P-gp after SE peaked at 48 h, which maybe the optimal detection time to detect drug resistant after SE.

Objective To investigate the value of therapeutic bandage type contact lens in the treatment of severe dry tear with dry eye. Methods 54 cases of severe tear evaporative dry eye patients from January 2016 to April 2017 were randomly divided into observation group (27 cases) and control group (27 cases). The observation group received the treatment of artificial tears combined with bandage contact lens treatment, the control group only received the treatment of artificial tears. Continuous treatment trials compared two groups of symptoms score, tear secretion after 1 months (SⅠt, SⅡt), tear break-up time (break up, time, BUT) and tear ferning test (tear ferning, test, TFT). Results After treatment, the scores of the two groups were decreased, and the observation group was lower than the control group. The difference was statistically significant (P＜0. 05); the levels of SⅠt, SⅡt and BUT were improved in both groups, and the improvement effect was better in the observation group (P＜0. 05); The detection level of TFT in the observation group was better than that in the control group, the difference was statistically significant (P＜0. 05). Conclusion The treatment of bandage contact lens in severe tear evaporation effect in treatment of patients with dry eye in strong type is effective and able to significantly improve their symptoms score and SⅠt, BUT and TFT index.

OBJECTIVE: To investigate the effect of exosomes derived from Epstein-Barr virus (EBV)-positive nasopharyngeal carcinoma (NPC) cells on lymphangiogenesis and lymph node metastasis of NPC. METHODS: Exosomes from NP69 cells and EBV-positive HK1 (HK1-EBV) cells were obtained by ultracentrifugation and identified by Western blotting and nanoparticle tracking analysis. Dio dye phagocytosis test was performed to observe exosome uptake by lymphatic endothelial cells. Lymphatic endothelial cells were treated with exosomes from nasopharyngeal epithelium (NP69), HK1-EBV, and C666-1 cells or exosome-free supernatant of HK1-EBV and C666-1 cells, and tube formation and migration of the cells were observed. In a nude mouse model of popliteal lymph node metastasis of NPC, the effects of normal saline, NP69 cell-derived exosomes, HK1-EBV cell-derived exosomes, exosome-free supernatant of HK1-EBV cells, and HK1-EBV exosome-free supernatant protein on lymphangiogenesis and lymph node metastasis of the tumor were observed. RESULTS: The exosomes obtained by ultracentrifugation contained abundant exosome-specific proteins and showed a normal size range. The exosomes from NPC cells and NP69 cells could be taken up by lymphatic endothelial cells. Compared with the blank control and exosomes form NP69 cells, exosomes derived from HK1-EBV and C666-1 cells significantly promoted tube formation and migration of lymphatic endothelial cells ( CONCLUSIONS Exosomes from EBV-positive NPC cells can significantly promote lymphangiogenesis and lymph node metastasis of NPC.
Animals ; Cell Line, Tumor ; Endothelial Cells ; Epstein-Barr Virus Infections ; Exosomes ; Herpesvirus 4, Human ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Mice ; Mice, Nude ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms

OBJECTIVE To explore the antitumor effect and mechanism of total alkaloids of Gelsemium elegans （TA） and sempervirine （SPV） in vitro. METHODS The effects of low， medium and high concentrations of TA （50， 100， 200 μg/mL） and SPV （10， 30， 50 μmol/L） on the morphology of human hepatoma cells （HepG2， Bel-7402）， human lung cancer cells （A549） and human colon cancer cells （HCT-8） were observed， and the toxicity of TA and SPV to four tumor cells was monitored. The effects of TA and SPV on the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of phosphorylated protein kinase B （p-Akt） （Thr308， Ser473）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， survivin， C/EBP-homologous protein （CHOP）， immunoglobulin binding protein （Bip） and microtubule-associated protein 1 light chain 3Ⅱ （LC3Ⅱ） in HCT-8 cells were detected. RESULTS After the intervention of TA and SPV， the volume reduction and nuclear shrinkage were founded in four tumor cells； the cell activity decreased to varying degrees， among which TA and SPV had the best inhibitory effect on HCT-8 cells. After the intervention of TA and SPV， the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of Bax， CHOP， Bip and LC3Ⅱ all increased to different degrees， while the protein expressions of p-Akt （Thr308， Ser473）， Bcl-2 and survivin in HCT-8 cells all decreased to different degrees. CONCLUSIONS TA and SPV have inhibitory effects on the above four tumor cells， and the inhibitory effect on HCT-8 cells is the best. The mechanism of their action on HCT-8 cells may be related to promoting apoptosis， activating endoplasmic reticulum stress and autophagy.