Objective To investigate the effect of valproic acid (VPA) on NKG2D-ligand expression in ARK,OPM2 human myeloma cell lines and their sensitization to natural killer (NK) cell-mediated Killing.Methods Different concentrations of VPA from 0-5.0 mmol/L were used to treat ARK,OPM2 cells respectively,then the cell viabilities were tested by flow cytometry (FCM).Real-time quantitative-PCR and FCM were used to detect the changes in mRNA,protein levels of NKG2D-ligand respectively in the two cell lines treated with 1 mmol/L VPA for 48 hours.The calcein-release-assay (CARE-LASS) was carried out to detect cytotoxic changes of NK cells against mydoma cells after VPA treatment.Results VPA induced the expression of MICA/B,ULBP2 (P ＜ 0.05) and in turn enhanced the NK cytotoxicity on myeloma cells.The enhancing effect of VPA was blocked by NK cells pretreated with anti-NKG2D mAb (P ＜ 0.05).The primary mechanism of NK cell killing of myeloma cells was perforin/granzyme-mediated.Conclusion VPA can induce the expression of MICA/B,ULBP2 in ARK,OPM2 cells,thereby enhancing the cytotoxicity against myeloma cells,which implies a new mechanism of anticancer approach and may be a new approach in myeloma immunotherapy.

OBJECTIVE To strengthen the management for preventing and controlling nosocomial infection.METHODS The normal work in this field according to the standards of Ministry of Health was summarized.RESULTS Changing the cognition of management of nosocomial infection to comprehend the meaning of management of nosocomial infection.CONCLUSIONS We can control nosocomial infection and ensure medical safety with the recognition of the leader,consummating organization,perfecting system,normalizing management,cooperating among departments,staff's effort and ensuring supply.

The purpose of this study was to clone human HSP90beta cDNA and construct its eukaryote expression vector . The total RNA was isolated by TRIzol Reagent (Invitrogen) from human NPC and its cDNA was gained by RT-PCR. The purified RT-PCR products and PGEM-T Easy Vector were ligated and transformed into XL1-blue E. coli bacteria. The white clones were selected and the plasmid was purified, which was further identified by double enzyme digestion and sequenced. PGEM-hHSP90beta and pcDNA3.1(+) DNA were digested by AflII and Xbal respectively. After purification, the two fragments obtained were ligased by using T4 DNA ligase (Fermentas). This recombinant DNA was then transformed into E. coli Competent Cells XL1-blue and positive clones were selected on the LB agarose plate containing Ampr (100 microg/ml). Single clones were identified by double digestion with AflII and Xbal, and two fragments with the size 5.4 kb and 2.1 kb were produced as expected. The hHSP90beta gene was successfully inserted into the eukaryote expression vector pcDNA3.1(+) by the recombination technique in vitro.
Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; HSP90 Heat-Shock Proteins ; biosynthesis ; genetics ; Humans ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Objective To preliminarily evaluate the effect of levocetirizine hydrochloride at different concentrations on the growth of in vitro cultured human dermal papilla cells,and to explore its mechanism.Methods Human dermal papilla cells were divided into several groups to be cultured with Dulbecco's modified eagle medium (DMEM) containing 0 (control group),1,10,100,1 000,10 000 μg/L levocetirizine hydrochloride respectively for 48 hours.Immunofluorescence staining was performed to observe the growth of the dermal papilla cells,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of the dermal papilla cells.Real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expression of cyclooxygenase 2 (COX-2),prostaglandin D2 synthase (PTGDS),prostaglandin E2 (PGE2),prostaglandin F2alpha (PGF2α),G protein-coupled receptor 44 (GPR44),protein kinase B (AKT) and glycogen synthase kinase 3β (GSK3β),and Western blot analysis to determine the protein expression of PTGDS.After 24-hour culture with DMEM containing levocetirizine hydrochloride at different concentrations,enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of prostaglandin D2 (PGD2) and PGD2R receptor in the culture supernatant of the human dermal papilla cells.Statistical analysis was carried out with SPSS 17.0 software using one-way analysis of variance for the comparison of the above indices among the groups,and least significant difference (LSD)-t test for multiple comparisons.Results Immunofluorescence staining showed that human dermal papilla cells grew well and reached over 90％ confluence in the 100 μg/L levocetirizine hydrochloride group.MTT assay revealed that there were significant differences in the proliferation rate among all the groups (F =42.22,P ＜ 0.05),and the proliferation rate was significantly higher in the 100 μg/L levocetirizine hydrochloride group (115.80％ ± 5.10％) than in the control group (100％,t =28.26,P ＜ 0.05).The mRNA expression(2-△△Ct) of COX-2,PGF2a,PTGDS,GPR44 and AKT all significantly differed among these groups (F =1.97,3.66,2.17,2.66 and 7.32 respectively,all P ＜ 0.05),while no significant difference in the mRNA expression of PGE2 and GSK3β was observed among these groups (F =0.87 and 1.19 respectively,both P ＞ 0.05).The 100 μg/L levocetirizine hydrochloride group showed significantly decreased mRNA expression of COX-2,PTGDS and GPR44 (0.84± 0.08,0.81±0.10 and 0.85 ± 0.09 respectively) compared with the control group (t =1.97,2.17 and 2.66 respectively,all P ＜ 0.05),but significantly increased mRNA expression of PGF2α and AKT (1.96 ± 0.25 and 1.74 ± 0.32 respectively) compared with the control group (t =3.66,7.32 respectively,both P ＜ 0.05).Moreover,the protein expression of PTGDS,PGD2 and PGD2R significantly differed among these groups (all P ＜ 0.05),and was significantly lower in the 100 μg/L levocetirizine hydrochloride group than in the control group (P ＜ 0.05).Conclusion Levocetirizine hydrochloride can promote the in vitro growth of human dermal papilla cells,likely by inhibiting the PGD2-GPR44 pathway and activating the AKT signal pathway.

Objective To explore the correlation of waist-to-height ratio (WHtR) and triglycerides (TG) with changes in cerebral hemodynamic index (CVHI) and the risk of stroke. Methods From March 2018 to March 2019, a total of 500 medical examinees were selected from the civil servants who underwent a physical examination in the physical examination department of our hospital. According to the TG level, the subjects were divided into two groups: HTG group (208 cases) and normal group (292 cases). According to WHtR value, the subjects were divided into an abnormal group (216 cases) and a normal group (284 cases). According to the test results, CVHI and other information the stroke risk was analyzed among the 4 groups. Results The right Vmax and bilateral Qmean, Vmean, Vmin, and Dp in the HTG group were significantly lower than those in the normal group (P<0.05), while the bilateral Wv, Zcv, Rv, DR, and Cp were higher than those in the normal group (P<0.05). The parameters of bilateral Vmax, Qmean, Vmean, Vmin, and Dp in the abnormal group were significantly lower than those in the normal group (P<0.05), while the bilateral Wv, Zcv, Rv, DR, and Cp were higher than those in the normal group (P<0.05). The CVHI scores of the HTG group and the abnormal group were lower than those of the normal group (P<0.05), and the FSP scores were higher than those of the normal group (P<0.05). The proportion of high-risk strokes with CVHI<75 and FSP score ≥10 in the HTG group were both higher than that in the normal group (P<0.05). There were 4 predictors including HTG, WHtR, CVHI score, and FSP score. There was a statistically significant difference in the etiology of stroke between CVHI and FSP (P<0.05). The risk of stroke in patients with abnormal WHtR increased by 2.746 times, and the relative risk of stroke in patients with CVHI<75 increased by 3.298 times. Conclusion WHtR abnormality and CVHI<75 were two independent predictors of stroke risk. HTG may increase the risk of stroke by affecting cerebral hemodynamic indicators.

Objective:To explore the application of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score in the timing and nursing of noninvasive ventilation for patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD).Methods:106 AECOPD patients admitted to Haikou People's Hospital from January 2018 to October 2019 were selected as the study objects. According to the method of random number table, the patients were divided into observation group and control group, with 53 patients in each group. The control group selected the timing of noninvasive ventilation treatment according to the standards of Mechanical ventilation (second edition), weaned according to Clinical practice of mechanical ventilation, and received routine nursing in intensive care unit (ICU), including creating comfortable indoor environment, reasonable diet, condition monitoring, psychological nursing and complications nursing. On the basis of the control group, the patients in the observation group were given noninvasive ventilation when APACHE Ⅱ score was more than 10, and were weaned when APACHE Ⅱ score was less than or equal to 10. According to APACHE Ⅱ score < 10, 10-14, 15-19 and ≥ 20, the patients were given level-3 care, level-2 care, level-1 care and intensive care. The pulmonary function before and 3 days after the noninvasive ventilation treatment was monitored, and the duration of mechanical ventilation, the length of ICU stay, endotracheal intubation rate, incidence of complication [ventilator associated pneumonia (VAP)] and ICU mortality were recorded. The self-designed questionnaire of nursing satisfaction was used to evaluate the patients' nursing satisfaction. Results:There was no significant difference in general data such as gender or age between the two groups. After 3 days of noninvasive ventilation, the forced expiratory volume in one second (FEV1), forced vital capacity (FVC) and FEV1/FVC ratio of the two groups were increased significantly as compared with those before treatment, especially in the observation group, with statistical significances as compared with the control group [FEV1 (L): 3.02±0.22 vs. 2.54±0.19, FVC (L): 3.01±0.32 vs. 2.13±0.28, FEV1/FVC ratio: 0.89±0.08 vs. 0.79±0.08, all P < 0.05]. Compared with the control group, the duration of mechanical ventilation and length of ICU stay in the observation group were significantly shortened [duration of mechanical ventilation (days): 4.32±0.73 vs. 8.42±1.94, length of ICU stay (hours): 32.23±10.22 vs. 38.52±9.85, both P < 0.01]. The intubation rate, incidence of VAP and ICU mortality in the observation group were significantly lower than those in the control group [intubation rate: 1.9% (1/53) vs. 13.2% (7/53), incidence of VAP: 1.9% (1/53) vs. 15.1% (8/53), ICU mortality: 1.9% (1/53) vs. 13.2% (7/53), all P < 0.05]. The nursing satisfaction of patients in the observation group was significantly higher than that in the control group [96.2% (51/53) vs. 75.5% (40/53), P < 0.01]. Conclusions:APACHE Ⅱ score can be used to guide the choice of noninvasive ventilation treatment opportunity and nursing intervention measures for AECOPD patients. It can significantly improve the pulmonary function of patients, improve the treatment effect, reduce the incidence of complications, and improve the satisfaction of patients with nursing, which is effective in clinical application.

Objective: To investigate the pathogen spectrum distribution and drug resistance of febrile neutropenic patients with hematological diseases in Shanghai. Methods: A retrospective study was conducted on the clinical isolates from the febrile neutropenic patients hospitalized in the departments of hematology in 12 general hospitals in Shanghai from January 2012 to December 2014. The drug susceptibility test was carried out by Kirby-Bauer method. WHONET 5.6 software was used to analyze pathogenic bacteria and drug susceptibility data. Results: A total of 1 260 clinical isolates were collected from the febrile neutropenic patients. Gram-positive bacteria accounted for 33.3% and Gram-negative bacteria accounted for 66.7%. Klebsiella pneumoniae (12.5%) , Stenotrophomonas maltophilia (9.5%) , Escherichia coli (9.1%) , Pseudomonas aeruginosa (8.7%) , Acinetobacter baumannii (6.6%) , Staphylococcus aureus (5.6%) and Enterococcus faecium (5.0%) were ranked in the first 7 of all pathogens. In the respiratory tract secretions specimens, non-fermented strains accounted for 56.2%. Stenotrophomonas maltophilia accounted for 15.2%. Enterobacteriaceae and coagulase-negative Staphylococci accounted for 42.3% (104/246) and 32.6% (85/246) respectively in blood samples. Enterobacteriaceae and Enterococcus bacteria accounted for 39.4% (76/193) and 28.5% (55/193) respectively in pus specimens. The detection rates of methicillin resistant Staphylococcus aureus (MRSA) and methicillin resistant coagulase negative Staphylococci (MRCNS) were 54.3% and 82.5%, respectively. Staphylococcus bacterial strain was not found to be resistant to linezolid, vancomycin and teicoplanin. The detection rate of Enterococcus vancomycin-resistant strains was 8.9%. Enterococcus was not detected resistance to oxazolidinone strains. Enterobacteriaceae bacteria were highly sensitive to carbapenems. The resistance rate of Pseudomonas aeruginosa to imipenem and meropenem was 34.1% and 15.8%, respectively. Stenotrophomonas maltophilia was more sensitive to minocycline hydrochloride, levofloxacin and sulfamethoxazole. The resistance rate of Acinetobacter baumannii only to cefoperazone-sulbactam was less than 10.0%. The antibiotic resistance rate of Klebsiella pneumoniae, Stenotrophomonas maltophilia, Pseudomonas aeruginosa and Acinetobacter baumanii to most of common antibiotics was lower than that of the CHINET surveillance. Conclusions The pathogenic strain distribution in common infection sites of febrile neutropenic patients was characterized. Bacterial resistance surveillance was better than the CHINET nationwide large sample surveillance in China.

Objective: To analyze the survival time of people living with HIV/AIDS and related influencing factors in Sichuan province during 1991-2017. Methods: A retrospective cohort study was conducted to analyze the data of 143 988 HIV/AIDS cases. The data were collected from Chinese HIV/AIDS Comprehensive Information Management System. Life table method was used to calculate the survival proportion of the cases, and Cox proportion hazard regression model was used to identify the factors related with survival time. Results: Among 143 988 HIV/AIDS cases a total of 30 420 cases died of AIDS related diseases (21.1%) and the average survival time was 11.51 years (95%CI: 11.39-11.64). Multivariate Cox regression analysis showed that the influencing factors for the survival of HIV/AIDS cases were gender (male vs. female, HR=1.35, 95%CI: 1.32-1.40), education level (primary school or below vs. junior middle school: HR=1.15, 95%CI: 1.12-1.18), ethnic group (Han vs. other ethnic groups, HR=1.46, 95%CI: 1.41-1.52), occupation (farmer vs. other occupations: HR=1.26, 95%CI: 1.22-1.29), age (≥55 years old vs. 15-24 years old: HR=3.18, 95%CI: 3.02- 3.36), disease phase (AIDS vs. HIV infection: HR=1.44, 95%CI: 1.39-1.48), antiretroviral therapy (ART) (receiving ART vs. receiving no ART: HR=0.20, 95%CI: 0.19-0.20), and CD₄⁺T cell counts at diagnosis (>500 cells/μl vs.<200 cells/μl: HR=0.42, 95%CI: 0.40-0.45). Conclusions The average survival time of HIV/AIDS cases was 11.51 years in Sichuan during 1991- 2017. The risk factors for the survival of the cases were male, education level of primary school or below, Han ethnic group, farmer, old age at diagnosis, disease phase, The protective factors for the survival of HIV/AIDS cases were receiving ART and higher CD₄⁺ T cell counts at diagnosis.

Tocilizumab has been reported to attenuate the "cytokine storm" in COVID-19 patients. We attempted to verify the effectiveness and safety of tocilizumab therapy in COVID-19 and identify patients most likely to benefit from this treatment. We conducted a randomized, controlled, open-label multicenter trial among COVID-19 patients. The patients were randomly assigned in a 1:1 ratio to receive either tocilizumab in addition to standard care or standard care alone. The cure rate, changes of oxygen saturation and interference, and inflammation biomarkers were observed. Thirty-three patients were randomized to the tocilizumab group, and 32 patients to the control group. The cure rate in the tocilizumab group was higher than that in the control group, but the difference was not statistically significant (94.12% vs. 87.10%, rate difference 95% CI-7.19%-21.23%, P = 0.4133). The improvement in hypoxia for the tocilizumab group was higher from day 4 onward and statistically significant from day 12 (P = 0.0359). In moderate disease patients with bilateral pulmonary lesions, the hypoxia ameliorated earlier after tocilizumab treatment, and less patients (1/12, 8.33%) needed an increase of inhaled oxygen concentration compared with the controls (4/6, 66.67%; rate difference 95% CI-99.17% to-17.50%, P = 0.0217). No severe adverse events occurred. More mild temporary adverse events were recorded in tocilizumab recipients (20/34, 58.82%) than the controls (4/31, 12.90%). Tocilizumab can improve hypoxia without unacceptable side effect profile and significant influences on the time virus load becomes negative. For patients with bilateral pulmonary lesions and elevated IL-6 levels, tocilizumab could be recommended to improve outcome.
Antibodies, Monoclonal, Humanized ; COVID-19/drug therapy* ; Humans ; SARS-CoV-2 ; Treatment Outcome