AIM: RNAⅢ inhibiting peptide (RIP) has been previously proved to possess good histocompatibility and safety for preventing and curing staphylococcal infection, and this study evaluated histocompatibility of polyaiticglycolic acid/RIP (PLGA/RIP) sustained release microsphere. METHODS: The experiment was performed in the Orthopedic Institute, Pharmacologic Research Institute and Animal Experimental Center of General Hospital of Chinese PLA from October 2005 to October 2007.①Preparation of PLGA/RIP: The solid-phase synthesis (Fmoc) method was used to synthesize RIP from C end to N end, then the synthesized peptide was purified by the reverse phase high performance liquid chromatography, and composition was collected by means of ultraviolet absorption peak. The purified RIP was obtained after freezing and drying. Liquid-phase multiple emulsion method was used to synthesize PLGA/RIP microsphere of 50-70 ?m diameter.②Acute general toxicity test was studied in PLGA/RIP. Effect of PLGA/RIP on the cell proliferation was detected with cytotoxicity test by MTT method. Intramuscular implanting test was used to observe the irritation reaction of muscles by implantation materials. Sensitivity test was used to observe the sensitization of PLGA/RIP. Changes of animal's body temperature were determined with pyrogen test. RESULTS: ①Acute general toxicity test: Neither toxicosis reaction nor animal death was found after animals were injected with 100% and 50% eluents of PLGA/RIP peritoneally. Animal's body weight was not changed significantly.②Cytotoxiciity test by MTT method: The average proliferation rate of cell in two kinds of eluents exceeded 85% and cytotoxicity was graded in 1 rank, indicating no cytotoxicity.③Intramuscular implanting test: At 4 weeks after RIP and PLGA/RIP were implanted into the animals, there was not obvious synathresis, denaturation or necrosis in tissues. No inflammatory cell infiltration occurred around the materials. There had been the fibrous capsules around the materials.④Sensitivity test: Average primary irritation index of three groups were 0.38, 0.33 and 0.31 respectively. There was no significant difference among three groups.⑤Pyrogen test: Fervescence of each animal in the experiment was under 0.5 ℃, confirming that the materials had no pyrogenic characteristics. This was in coincidence with evaluation criterion of pyrogen test. CONCLUSION: PLGA/RIP has good histocompatibility and safety, without general toxic reaction, cytotoxicity, immunological rejection, hypersensitive response or pyrogenic characteristics.

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Objective To study the effect of Zhimaikang Granules (Granules for lipid abnormality) on atherosclerosis of hyperlipidemia patients of phlegm turbidity retention type.Methods Totally 120 hyperlipidemia patients were randomized into treatment group and control group, 60 cases in each. The treatment group was administered Zhimaikang Granules, each time 1 bag, twice a day. The control group was given Simvastatin Tablets, oral taking before sleeping. Both groups were treated for 45 days. Changes of superoxide dismutase (SOD), malondialdehyde (MDA), and nitric oxide (NO) were observed.Results After treatment, activity of NO and SOD were significantly higher than before in both groups (P0.05).Conclusion Zhimaikang Granules was proved to enhance the activity of antioxidase, promote anti-free radical reactions, inhibit lipid peroxidation and protect endothelial cells in atherosclerosis patients.

Objective To explore the potential impact of low concentration of metformin on the morphology and function of mitochondria of HepG2 cells. Methods HepG2 cells in experimental group and control group were treated with or without low concentration of metformin (1mM/L), respectively. The cells were incubated for 12h in the incubator with constant temperature and humidity as well as 1% oxygen. Orange Mitoview was used to stain the mitochondria to detect the effects of the drug on its morphology and quantity. Transmission electron microscope was utilized to observe the effect of metformin on the ultrastructure of mitochondria. The mitochondrial respiratory chain complex I activity in HepG2 cell was detected by Complex I Enzyme Activity Dipstick Assay Kit (DAK). Results Orange Mitoview staining showed that low concentration of metformin had little effect on the morphology and number of mitochondria of cells in experimental group , and the difference between control and experimental group was not statistically significant (P > 0.05). In addition, the result was further determined by transmission electron microscopy. However, DAK analysis showed that complex I activity of cells in experimental group was significantly lower than that in control group. Conclusion Under Hypoxia conditions, low concentration of metformin had no significant effect on the morphology and number of mitochondria of HepG2 cells, but it significantly reduces the activity of mitochondria of HepG2 cells.

OBJECTIVETo investigate the effect of PIK3CA/siRNA chitosan nanoparticle on the invasiveness of gastric carcinoma and the potential value of PIK3CA/siRNA chitosan nanoparticle in suppressing the metastasis of gastric carcinoma.

METHODSGastric cancer cells were treated with PIK3CA/siRNA nanoparticle (with a diameter of 350 nm), and the efficiency of PIK3CA gene interference was evaluated using Western blotting and real-time PCR. The changes of the invasive capacity of the treated cells was assessed with Transwell assay.

RESULTSPIK3CA/siRNA chitosan nanoparticle efficiently lowered the expression level of PIK3CA and significantly decreased the invasion of BGC823 cells.

CONCLUSIONPIK3CA gene interference mediated by PIK3CA/siRNA chitosan nanoparticle can decrease the invasive capacity of gastric cancer cells in vitro.

Cell Line, Tumor ; Cell Proliferation ; Chitosan ; Class I Phosphatidylinositol 3-Kinases ; Humans ; Nanoparticles ; Phosphatidylinositol 3-Kinases ; genetics ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Stomach Neoplasms ; pathology

Objective:To analyze the changes of serum lipidomics in patients with sepsis and healthy controls, search for the differences of lipid metabolites, and reveal the changes of lipidomics in the process of sepsis.Methods:A prospective observational study was conducted. From September 2019 to April 2020, morning blood samples of upper extremity superficial veins were collected from 30 patients with definite sepsis diagnosed in intensive care unit (ICU) of Shanxi Bethune Hospital and 30 age-matched healthy subjects during the same period. Serum lipid metabolites were analyzed by ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS), and the quality control samples were analyzed by base peak spectroscopy (BPC) and verified experimental repetition. Student t-test and fold change (FC) were used for screening significant differences in lipid metabolites and determining their expression changes. Principal component analysis (PCA) and orthogonal projectionto latent structure discriminant analysis (OPLS-DA) were used to determine the entire allocation of experimental groups apiece, access the quality of being near to the true value of model, and screen the differential lipid metabolites with variable importance of projection (VIP). Finally, Metabo Analyst platform database was used to analyze lipid molecular metabolic pathways. Results:BPC results showed that the experimental repeatability was good and the experimental data was reliable. The main parameter model interpretation rate of PCA model R 2X = 0.511, indicating that the model was reliable. The main parameter model interpretation rate of OPLS-DA model R 2Y = 0.954, Q 2 = 0.913, indicating that the model was stable and reliable. With FC > 2.0 or FC < 0.5, P < 0.05, a total of 72 differential lipid metabolites were obtained based on VIP > 1. Based on Metabo Analyst 5.0, 24 distinguishable lipid metabolites were identified including 8 phosphatidylethanolamine (PE), 7 lysophosphatidylcholine (LPC), 6 phosphatidylcholine (PC), 2 lysophosphatidylethanolamine (LPE) and 1 phosphatidylserine (PS). Compared with healthy volunteers, the lipid molecules expression proved down-regulated in most sepsis patients, including PC, LPC, LPE, and some PE, while some PE and PS were up-regulated, which was mainly related to the PE (18∶0p/20∶4), PC (16∶0/16∶0) and LPC (18∶1) metabolic pathways in glycerophospholipids. Conclusions:There are significant differences in lipid metabolites between the sera of sepsis patients and healthy volunteers. PE (18∶0p/20∶4), PC (16∶0/16∶0) and LPC (18∶1) may be new targets for sepsis prediction and intervention.

Objective To explore the correlation between platelet function and immunity index in patients with sepsis. Methods The platelet function and immune indexes of one hundred and one patients with sepsis treated in Shanxi Dayi Hospital from July 1st, 2016 to October 31st, 2017 were analyzed retrospectively. According to their shock,they were divided into shock group (34 cases) and non shock group (67 cases). Another 50 healthy people in the same period in our hospital were selected as control group. The relationship between platelet function and immune indexes was compared. Results ( 1) the incidence of maximum blood block intensity decreased in the thrombus map of the septic shock group was higher than that in the non shock group, and the difference was statistically significant ( 65. 67%( 44/67 ) vs. 23. 53%(8/34),χ2＝41. 28,P＜0. 05); (2) the CD4+T lymphocyte and C3 in the septic shock group were all lower than those in the non shock group ((47. 28%±7. 78) vs. (54. 93%±11. 26),t＝3. 554,P＜0. 05; (0. 42 ±0. 23) g/L vs. (0. 75±0. 19) g/L,t＝－3. 057,P＜0. 05),the ratio of CD4+/CD8+T lymphocyte was higher than that in non shock group ((2. 68±0. 18) vs. (2. 45±0. 07),t＝7. 18,P＜0. 001)). (3) the maximum intensity of blood clots was correlated with the percentage of CD4+T lymphocyte,CD4+/CD8+T lymphocyte ratio,complement C3,acute physiology and chronic health status score system II score,and sequential organ failure score ( r ＝ 0. 617, 0. 411, 0. 563,－ 0. 631,－ 0. 547, P＜ 0. 01, or P＜ 0. 05 ) . Conclusion Thrombocytopenia is present in septic patients,which is correlated with changes in immune indices.

Objective:To evaluate the prognostic value of thromboelastography maximum amplitude(MA)and arterial blood lactate levels for sepsis in elderly patients.Methods:A retrospective analysis was performed on clinical data of 63 sepsis patients(≥60 years old)admitted to the Intensive Care Unit(ICU)of Bethune Hospital of Shanxi Province from December 2018 to February 2020.MA values, white blood cell counts, lymphocyte counts, platelets, acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)scores, sequential organ failure assessment(SOFA)scores, underlying diseases, body mass index, laboratory test results and other related treatments were analyzed.The subjects were divided into the survival group and the death group according to the 28-day survival outcome.Differences in MA, APACHE Ⅱ scores, SOFA scores and laboratory test results between the two groups were analyzed, and the correlations of MA with infection parameters and age were examined.Influencing factors of survival outcomes were analyzed using multivariate Logistic regression.The receiver operating characteristic curve(ROC)was used to calculate the prognostic value of MA and arterial lactate for sepsis in elderly patients.Results:The main sources of infections were pulmonary and abdominal(79.4%, 50/63)in 63 elderly patients with sepsis.The incidences of positive blood cultures and deaths were 15.9%(10/63)and 66.7%(42/63), respectively.There existed significant differences in lymphocyte counts, arterial lactate levels, MA and lengths of stay in the ICU between the survival group and the death group( t=3.847, 2.153, 2.745, -3.574, respectively, all P<0.05).MA was correlated with arterial lactate, SOFA score and survival outcome( r=-0.498, -0.506, and -0.358, respectively, all P<0.05).Multivariate Logistic regression analysis showed that MA and arterial lactate were independent factors for the survival outcome( OR=1.626, 0.766, all P<0.05).The area under the ROC curve(AUC, 95% CI)for the combination of MA and arterial lactate was larger than that of either MA or arterial lactate alone(0.89, range: 0.763-0.846; 0.58, range: 0.574-0.730; 0.77, range: 0.521-0.832; all P<0.05). Conclusions:The combination of thromboelastography maximum amplitude and lactate in arterial blood has important clinical value in assessing the prognosis of elderly patients with sepsis.

Objective:To screen lipid biomarker in sepsis patients with different survival outcome based on ultra high performance liquid chromatography-mass spectrometry(UHPLC-MS/MS) technique.Methods:From September 2019 to April 2020, 30 septic patients admitted in Department of Intensive Care Unit and 30 cases of physical examination at the same time in Shanxi Bethune Hospital were studied. Lipid metabolite in serum were detected by UHPLC-MS/MS technique. According to the 28 day survival outcome of sepsis patients, they were divided into survival group (21 cases) and death group (9 cases). The baseline data of case group and control group, survival group and death group were compared respectively. Independent sample t-test and orthogonal partial least squares discriminant analysis (OPLS-DA) were further performed to identify lipid biomarkers related to sepsis survival outcome. Receiver operating characteristic (ROC) curve to evaluate the predictive efficacy of differential lipids on the survival outcome of biomarker sepsis patients. Results:There were 32 lipid subclasses and 1 437 differential lipid molecules in the sepsis group compared with the control group. 196 differential lipid molecules in the sepsis survival group and the death group were screened according to the OPLS-DA model (variable weight of projection (VIP)>1), which were glycerophosphingolipids (129), sphingolipids (52), glycerides (14), and sterols (1).All the original data were statistically analyzed by univariate independent sample t-test. There were statistically significant differences in 15 lipid molecules between the two groups. Combined with VIP > 1 and P < 0.01, three lipid molecules were finally screened, which were sphingomyelin (SM) lipid molecules, SM (d30∶1), SM (d32∶2), SM (d32∶1). ROC curve analysis showed that the areas under curves of the above three lipid molecular were 0.915, 0.892, 0.898, respectively. The sensitivity was 77.27%, 95.45%,72.73%. The specificity was 100.0%, 87.5%,100.0%. Further Z-test showed that there was no significant difference in the area under the ROC curve ( Z(SM (d30∶1) and SM (d32∶1)) =0.36, P=0.722; Z(SM (d30∶1) and SM (d32∶2))=0.34, P=0.732; Z(SM (d32∶1) and SM (d32∶1))=0.07, P=0.942). Conclusions:Sphingomyelin may be involved in the formation of different clinical outcomes of sepsis, and has a good predictive effect on the survival outcome of sepsis.

Objective: To study the effect of low-to-moderate dose glucocorticoid therapy on viral clearance time in patients with COVID-19. Methods: A total of 72 patients diagnosed with COVID-19 from January 19 to February 17, 2020 at the First Affiliated Hospital, School of Medicine, Zhejiang University were recruited. All patients received oral abidol and/or combined lopinavir/ritonavir, darunavir antiviral, and symptomatic supportive care. Among them, 51 patients received methylprednisolone (0.75-1.50 mg·kg^-1·d^-1) (glucocorticoid treatment group), and 21 patients who did not use glucocorticoid were the control group. The time of stable virologic conversion insputumand the time of radiologic recovery in lungsince onset were compared between the two groups and among the normal patients.The Kruskal-Wallis test or Fisher exact test was used to compare the difference between groups. Results: The median ages of the glucocorticoid group and the control group were 52 [interquartile range (IQR):45, 62] years and 46 (IQR: 32, 56)years, and the differences were significant (P<0.05). The clinical conditions at hospital admission were different between the two groups (P<0.01). There were 52.0% critical ill patients in the glucocorticoid treatment group, compared to that of 71.4% normal patients in the control group. The median times from the onset tostable virologic conversion to negative in the two groups were 15 (IQR:13,20) days and 14 (IQR:12,20) days (P>0.05), and the difference was no statistically significant. The median times from onset to radiologic recovery were 13 (IQR: 11,15) days and 13 (IQR:12,17) days in the two groups, and there was no difference (P>0.05). In ordinary patients, the median timesfrom the onset tostable virologic conversion insputum were no difference (P>0.05), with 13 (IQR:11,18) days in the glucocorticoid group and 13 (IQR:12,15) days in the control group; The median times from onset to radiologic recovery in lungwere also no difference (P>0.05), with 12 (IQR: 10,15)days in the glucocorticoid group and 13 (IQR: 12,17) days inthe control group. Conclusions Low-to-moderate glucocorticoid treatment has no effect on the time of virus clearance in patients with different clinical types of COVID-19. The glucocorticoid is not recommended since no effectiveness on accelerating the improvement of radiologic recovery in lung has been observed.