Status of tranquility nursing in Jinshan District， Shanghai / 上海预防医学

Objective:To analyze current situation of hospice care in Jinshan District of Shanghai and provide recommendations for development of hospice care. Methods:This cross-sectional study included 8 hospice care institutions， their health staff and 316 patients during May-August， 2019. Results were analyzed by quantitative and qualitative methods. Results:There had 80 approved beds in Jinshan District， but only 53 beds were used for 189 inpatients in 2017. Hospice care institutions received financial supports from district government at the construction stage， but relied on their own for daily operation costs. The motivation of health staff to work in hospice care was not strong. Among the hospice care workers， 18% had no certification for hospice care， and only 5% of them had senior professional titles. Continued education for health workers in hospice care was weak and the workers were under heavy stress. Conclusion:The potential needs for hospice care were unmet while hospice care was under-utilization in Jinshan District. The development of hospice care was unbalanced， and the health workers lacked motivation. The resources of hospice care need to be strengthened.

Exosomes from bone marrow mesenchymal stem cells regulate the balance of Foxp3+ Treg/Th17 in asthmatic mice / 中国组织工程研究

BACKGROUND: Imbalance of Th1/Th2 immune response is an crucial pathophysiological manifestation of asthma, but recent studies have proved that asthma also has a close correlation with the imbalance of Foxp3+Treg/Th17. Accumulating evidence indicate that the immunoregulatory capacity of mesenchymal stem cells are mainly related to exosomes secreted by the cells. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cell exosomes on Foxp3+Treg cells, Th17 T cells and airway inflammation of asthmatic mice as well as cytokines in the bronchoalveolar lavage fluid. METHODS: Twenty-seven BALB/c mice of SPF grade were randomly divided into three groups: normal control group, asthmatic model group, and exosomes group. Except the normal control group, each mouse in the other groups was sensitized by ovalbumin to establish asthma models. In the exosomes group, each mouse was intravenously administrated with exosomes at 21 days of sensitization. At 24 hours after the final administration of ovalbumin, the proportion of Foxp3+Treg and Th17 in the sleep of asthmatic mice as well as the expression of CTLA-4 and PD-1 in Foxp3+Treg cells were detected by flow cytometry. The total number of inflammatory cells, and the number of eosinophils, lymphocytes, monocytes and neutrophils in the bronchoalveolar lavage fluid were counted to analyze the degree of airway inflammation in the combination with pathological observation. We also detected the expression of interleukin-4,5,13,10,17 and interferon-γ in the bronchoalveolar lavage fluid as well as p27kip1in CD4+T cells. RESULTS AND CONCLUSION: (1) The proportion of Foxp3+Treg in splenic lymphocytes and the CTLA-4 and PD-1 expression in Foxp3+Treg were significantly higher in the exosomes group than the asthmatic model group (P < 0.01). (2) The proportion of Th17 in splenic lymphocytes was ranked as follows: asthmatic model group > exosomes group > normal control group (P < 0.01). (3) The total number of inflammatory cells and the number of eosinophils, lymphocytes, neutrophils and monocytes in the bronchoalveolar lavage fluid were ranked as follows: asthmatic model group > exosomes group > normal control group (P < 0.01). (4) Pathological observation of the lung showed that the asthmatic mice appeared to have severest airway inflammation. However, mesenchymal stem cell exosomes could significantly alleviate the airway inflammation. (5) The detection of cytokines in the bronchoalveolar lavage fluid showed that levels of interleukin 13 and interleukin 17 were significantly reduced (P < 0.05, P < 0.01), while the level of interleukin 10 increased (P < 0.01). (6) The p27kip1expression in the CD4+T cells was obviously higher in the exosomes group than the asthmatic model group. In conclusion, exosomes from bone marrow mesenchymal stem cells can reverse the imbalance of Foxp3+Treg/Th17 and significantly inhibit the airway inflammation in asthmatic mice.

Effect of regulating platelet function on systemic inflammatory response and pul-monary oxygenation function in rats during extracorporeal circulation / 中国免疫学杂志

Objective:To investigate the effect of regulating platelet function on systemic inflammatory response and pulmonary oxygenation function in rats during extracorporeal circulation(ECC).Methods:The rats participating in the ECC model were randomly divided into ECC supplemented platelet group (group MH),ECC supplemented plasma group (group MP),and continuous pump Tirofiban+ECC supplemented platelet group (group TMH),with 8 rats in each group.Rats of the three groups lasted two hours with ECC,observed for two hours after operation.Enzyme linked immunosorbent assay (ELISA) was used to detect inflammatory factors in the plasma and bronchoalveolar lavage fluid of rats in the three groups.The oxygenation index and lung tissue water content were meas-ured.The number of circulating endothelial quantity was measured by flow cytometry,and pathological changes of lung tissue in rats were observed by hematoxylin eosin staining.The effects of regulating platelet function on the levels of systemic inflammatory response, the degree of vascular endothelial damage,the edema degree of lung tissue and pulmonary oxygenation function of ECC rats were analyzed.Results:There was no significant difference in platelet count between group MH and group MP after two hours with ECC(P>0.05),the group TMH was significantly higher than that in group MH and group MP (P<0.05).After two hours with ECC,the oxygenation indexes of the three groups were lower than that at the beginning of ECC(P<0.05).There was no significant difference in lung tissue water content between the three groups(P>0.05);the level of neutrophil elastase(NE) in bronchoalveolar lavage fluid of rats in group MH was lower than that in group TMH(P<0.05),the level of NE in plasma of group TMH was lower than that in group MP (P<0.05).At the end of ECC,there were no statistically significant differences in circulating endothelial quantity of the three groups (P>0.05).Pathological examination of lung tissue showed that the lung interstitium of group MH was thinner than that of other groups,and inflammatory cell infiltration was less.Conclusion:At the early stage of ECC,supplement fresh platelets after platelet sharp decrease can significantly reduce the inflammatory response in the circulation.The protective effect may be related to platelets adhere to the endothelium associated with ECC related inflammatory reactions,and decrease the neutrophils and endothelial tissue adhesion.It is suggested that certain number of platelets with normal adhesion and aggregation function in ECC may reduce inflammatory response and lung injury after operative,and improve prognosis.

Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 in vitro / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the expression of connective tissue growth factor (CTGF) and α-SMA in human lens epithelium cell (HLEC) line B3 after transfection by liposome-coated siRNA targeting CTGF.METHODS: HLECs were transfected with small interfering RNA (siRNA) targeting CTGF,labeled with 5`-fluorescein isothiocyanate (5`-FITC) and coated with lipofectamine.The transfection ratio was evaluated via fluorescence intensity.Cell counting kit-8 (CCK-8) assay was performed to assess cytoviability of both non-transfected and transfected HLECs.Quantitative RT-PCR,cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection.RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine.The transfection ratio reached 95% at 24h.The proliferation of HLECs was inhibited by siRNA after 72h transfection.The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h.This effect was not found in negative control siRNA.CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs,which is a potential therapeutic strategy for posterior capsular opacification.

Optimal Threshold of Stimulated Serum Thyroglobulin Level for 18F-FDG PET/CT imaging in Patients with Thyroid Cancer / 华中科技大学学报（医学）（英德文版）

This study was to explore the optimal threshold of thyroid-stimulating hormone (TSH)-stimulated serum thyroglobulin (s-Tg) for patients who were to receive 18F-fluorodeoxyglucose (18F-FDG) PET/CT scan owing to clinical suspicion of differentiated thyroid cancer (DTC) recurrence but negative post-therapeutic 131I whole-body scan (131I-WBS).A total of 60 qualified patients underwent PET/CT scanning from October 2010 to July 2014.The receiver operating characteristic (ROC) curve analyses showed that s-Tg levels over 49 μg/L led to the highest diagnostic accuracy of PET/CT to detect recurrence,with a sensitivity of 89.5％ and a specificity of 90.9％.Besides,bivariate correlation analysis showed positive correlation between s-Tg levels and the maximum standardized uptake values (SUVmax) of 18F-FDG in patients with positive PET/CT scanning,suggesting a significant influence of TSH both on Tg release and uptake of 18F-FDG.So,positive PET/CT imaging is expected when patients have negative 131I-WBS but s-Tg levels over 49 μg/L.

TIPS versus Endoscopic Therapy for Variceal Rebleeding in Cirrhosis:A Meta-analysis Update / 华中科技大学学报（医学）（英德文版）

Endoscopic therapy (ET) is most common method for preventing variceal bleeding in cirrhosis,but the outcomes are not perfect.Recently,transjugular intrahepatic portosystemic shunt (TIPS) is introduced into clinical practice.However,the beneficial effects of TIPS compared to ET on cirrhotic patients is unknown.The aim of this study was to evaluate and compare the effects of TIPS with those of the most frequently used ET for prevention of variceal rebleeding (VRB) in liver cirrhosis.The PubMed,EMBASE,and Cochrane Library databases were searched from inception to February 2017.The primary study outcomes included the incidence of VRB,all-cause mortality,bleeding-related death,and the incidence of post-treatment hepatic encephalopathy (PTE).The odds ratios (ORs) with 95％ confidence intervals (CI) were pooled for dichotomous variables.Subgroup analyses were performed.Twenty-four studies were eligible and they included 1120 subjects treated with TIPS and 1065 subjects treated with ET.Although there was no significant difference in survival and PTE,TIPS was superior to ET in decreasing the incidence of VRB (OR=0.27;95％ CI,0.19-0.39,P＜0.00001),and decreasing the incidence of bleeding-related death (OR=0.21;95％ CI,0.13-0.32,P＜0.00001).Subgroup analysis found a lower mortality (OR=0.48;95％ CI,0.23-0.97;P=0.04) without any increased incidence of PTE (OR=1.37;95％ CI,0.75-2.50;P=0.31) in the studies of a greater proportion (≥40％) of patients with Child-Pugh class C cirrhosis receiving TIPS,and TIPS with covered stent did not increase the risk of PTE compared to ET (OR=1.52,95％ CI =0.82-2.80,P=0.18).It was concluded that TIPS with covered stent might be considered the preferred choice of therapy in patients with severe liver disease for secondary prophylaxis.

Efficacy of EGFR tyrosine kinase inhibitors in advanced non-small cell lung cancer patients with uncommon EGFR 21L861Q mutation / 上海交通大学学报（医学版）

Objective· To evaluate the efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKI) in non small cell lung cancer (NSCLC)patients with uncommon EGFR 21L861Q mutation. Methods·Between June 2011 and Marth 2015,clinical data of 21 stageⅢB/ⅣNSCLC patients who received EGFR-TKI harboring uncommon 21L861Q mutation in EGFR at the Shanghai Chest Hospital were collected.Objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS ) and overall survival (OS ) of the patients under TKI therapy were retrospectively analyzed. Results · ORR and DCR of the patients under TKIs therapy (first-line+second-line+third-line) were 42.9% and 66.7% respectively. PFS and OS of patients who received therapy that consisted of EGFR-TKIs(first-line+second-line+third-line)were 7.03 months(95% CI,5.50-8.69)and 22.80 months(95% CI,16.22-25.65). Conclusion·Our post-hoc analyses demonstrated that EGFR-TKIs showed activity in patients with uncommon EGFR 21L861Q-mutant NSCLC, less effective than in those with common mutations.

The differentiation of human multipotent adult progenitor cells into hepatocyte-like cells induced by coculture with human hepatocyte line L02

PURPOSE: The aim of this study was to establish an in vitro method to purify human multipotent adult progenitor cells (hMAPCs) and assess their possible differentiation into hepatocytes by coculture with human hepatocyte line L02. METHODS: hMAPCs were isolated by magnetic activated cell sorting (MACS) depletion selection using CD45 and GlyA microbeads. After indirect or direct coculture of hMAPCs and human hepatocyte line L02, the expression of albumin (ALB), alpha-fetoprotein (AFP), cytokeratin (CK) 18, and CK19 by hMAPCs was detected by immunocytochemistry. RESULTS: With the MACS method, (5-10) x 10(4)/mL hMAPCs could be separated from 1 x 10(6)/mL bone marrow mononuclear cells. The purity of CD45-/GlyA- cells separated from bone marrow adherent cells was more than 98%, as determined by flow cytometry. In the coculture without cell-to-cell contact, hMAPCs expressed high AFP on day 1, and then tapered daily to low expression on day 7; ALB expression reached its peak on day 5, and remained high on day 7; CK18 was initially expressed on day 5 and was higher on day 7; CK19 was negative in all assays. In the coculture with cell-to-cell contact, ALB and CK18 were expressed by most cells while AFP appeared in only a few on day 5. CONCLUSION: hMAPCs were induced to differentiate into mature hepatocyte-like cells by coculture with a hepatocyte cell line, either with or without cell-to-cell contact, but the former seemed more effective.

Expression of interleukin-17 and retinoid-related orphan nuclear receptor gamma t in cornea of experimental keratomycosis / 中华实验眼科杂志

Background In the past few decades,the balance of Th1/Th2 is often used to explain the immune mechanisms of fungal infection and fungal disease.More recently,a novel subset of CD4+ effector Th cells has been found to participate in anti-fungal infection response.However,whether Th17 is involved in the immune response in fungal keratitis is unclear up to now.Objective Present study was to investigate the expression change of Th17 type cytokine and its specific transcription factor,retinoid-related orphan nuclear receptor gamma t (RORγt),in the cornea of Fusarium solani keratitis.Methods Ninety-six clean BALB/c mice were divided into Fusarium solani keratitis model group and control group by randomized digital table.Fusarium solani keratitis models were established by epikeratophakia-assisted corneal epithelial erasion and interlayerly injection of 5 μl (1 × 106 CFU/ml) Fusarium solani solution in the right eyes,and the equal volume of PBS was injected in the same way in the control group.10％ KOH wet film was used to examine the fungal hyphea and funga strain was identified by inoculation.The corneas were examined under the slit lamp microscope 1 day,3,5,7 days after modeling and the inflammatory response was scored based on the criteria of Wu and Hu.The histopathological examination of corneas was performed in the time points above.Real time fluorescence quantitative PCR was used to detect the expression levels of interleukin-17 (IL-17) mRNA and RORγt mRNA in the corneas.The expression of IL-17 protein in the corneas was detected by ELISA.The use and raise of the mice followed the Statement of Association for Research in Vision and Ophthalmology.Results The inflammatory scores were 3.2±0.8,6.6± 1.1,9.4± 1.1 and 6.8±0.8 in 1 day,3,5,7 days after modeling,showing a significant difference among them (F =89.786,P =0.010).The inflammatory scores were higher in the third and seventh day than that in the first day (P＜0.05),but they were significantly lower than that in the fifth day (P＜0.05).The infiltration of inflammatory cells showed a coincident tendency with the score.The expressing levels of IL-17 mRNA (2-ΔΔCt) in the corneas were 4.12±0.73,20.72±1.81 and 14.16±1.88 in 3,5,7 days after modeling,with statistically significant differences in comparison with those in the control group (P＜0.01),and the expression level was significantly higher in the fifth day than those in the first,third and seventh day in the model group(P＜0.01).The expression levels of IL-17 protein (ng/g) were significantly increased 1 day,3,5,7 days in the model group compared with the control group (P＜0.01).A similar change was found in the expression of RORγt mRNA to that of IL-17 mRNA.Conclusions Expressions of IL-17 and its transcription factor RORγt upregulate in the fungal keratitis and has an association with inflammatory degree,which suggests that Th17 subset may play an important role in the immune responses of fungal keratitis.

Establishment and characterization of a human gallbladder carcinoma cell line EH-GB1 originated from a metastatic tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To establish a human gallbladder carcinoma cell line derived from a metastatic gallbladder carcinoma and identify its biological characteristics.</p><p><b>METHODS</b>Tissue samples were separated from the surgical specimen obtained from a patient with metastatic carcinoma and single-cell suspension was prepared. Then the cells were cultured in DMEM medium supplemented with 15% fetal bovine serum. The morphology of tumor cells was observed under an electron microscope. The cell growth curve was plotted. The tumorigenicity of the cell line was studied by subcutaneous inoculation in SCID mice. The cells were infected by lentiviral vector carrying fluorescent report genes (lenti-GFP and lenti-Red2) separately for expressions of GFP and Red2, respectively.</p><p><b>RESULTS</b>A novel metastatic gallbladder carcinoma cell line was successfully established and named "EH-GB1". It could be passaged for over 20 generations with typical malignant epithelial morphology and a stable growth cycle of 24 h. Tumors were formed in all of the 10 SCID mice inoculated with EH-GB1 cells subcutaneously, and the tumor cells were tumor marker CA19-9-positive. Continuous expressions of fluorescent report genes were observed in EH-GB1 cells infected by lenti-GFP and lenti-Red2.</p><p><b>CONCLUSION</b>EH-GB1 cells might be the first stable cell line of human gallbladder carcinoma established from a metastatic focus of gallbladder carcinoma. This cell line with continuous expressions of GFP and Red2 might be a novel and perfect experimental model for clinical and basic research on gallbladder carcinoma.</p>

Preparation and biological activity of poly (gamma-glutamic acid) -cisplatin conjugate / 药学学报

Preparation of a poly (gamma-glutamic acid)-cisplatin conjugate was introduced and its in vitro antitumor effect was investigated. Poly (gamma-glutamic acids) was obtained by using fermentation methods. The hydrolyzed small molecular weight of poly (gamma-glutamic acids) was prepared by acid hydrolysis. The interaction between poly (gamma-glutamic acids) -cisplatin conjugate (PGA-CDDP) and DNA was investigated by PCR model. MTT assay was used to investigate the in vitro anticancer activity of the conjugate. Apoptosis assay of the conjugate was investigated by FCM assay and the in vivo toxicity was also proceeded. The results showed that the poly (gamma-glutamic acids) -cisplatin conjugate was obtained successfully and its yield is 10% - 12%. It has obvious antitumor effects on human liver tumor BEL7404 cells, human lung tumor H446 cells and human colon tumor RKO cells. At the same time, it also has apoptosis effects on the three kinds of tumor cell lines. The in vivo toxicity of PGA-CDDP was examined in normal mice and the results showed that the in vivo toxicity of this conjugate was significantly lower than that of free CDDP. In conclusion, the poly (gamma-glutamic acids) -cisplatin conjuate could be used as a potential clinic antitumor drug. The poly (gamma-glutamic acids) obtained by fermentation can be used as a valuable drug carrier system.

Advances in research of okadaic acid / 中国海洋药物

The origin,distribution,chemical structure and biological properties of okadaic acid are described in this paper.Research prospects of okadaic acid and several research fields with priority are proposed according to the present research progress in China.