Entorhinal Cortex/physiology* ; Hippocampus/physiology* ; Neural Pathways/physiology*

OBJECTIVE：To study the skin irritation and se nsitization of domestic generic drug Clobetasone butyrate cream ， and to compare it with commercial drug （original drugs ）. METHODS ：The skin irritation test was conducted on rabbits. Totally 24 rabbits were randomly divided into test preparation intact skin group ，test preparation abraded skin group ，commercial drug intact skin group and commercial drug abraded skin group ，with 6 rabbits in each group. 0.5 mL test preparation or commercial drug was administered to the left side of intact or abraded skin and the same amount of excipient on the right side of each rabbit twice a day for consecutive 7 days. The irritation of the drug to the rabbit skin was observed ，and the erythema and edema of the skin were scored；the skin of administration site was taken at 72 h after last administration and the end of 7 d after drug withdrawal for histopathological examination. The skin sensitization test （Buehler test ）was carried out on guinea pigs. Totally 60 guinea pigs were randomly divided into test preparation group （n＝20），commercial drug group （n＝20），positive control group （n＝10）and excipient control group （n＝10）. 0.2 mL test preparation or commercial drug was administrated to the left side of the rib abdomen skin of each guinea pig at the 0，7th，14th day to induce model ，and an equal amount of corresponding preparation was administered to the right side in the same way at the 28th day for stimulation. Hypersensitive response such as erythema and edema were observed and scored at 24 h and 48 h after the stimulation. The incidence of hypersensitive response was then calculated. RESULTS：In skin irritation test of rabbits ，no erythema and edema was caused by the test preparation or commercial drug on intact skin of rabbits ；scores of skin irritation was 0；there was no dermal irritation. Both test preparation and commercial drug caused transient slight erythema on abraded skin of a few rabbits ；scores of intact and abraded skin irritation were 0-0.33；there was no dermal irritation. There was no statistical significance among groups. No dermal pathological changes were observed. In skin sensitization test of guinea pig ，no hypersensitive response such as erythema and edema was found on the skin of guinea pigs in both test preparation and commercial drug groups ；both score and the incidence of hypersensitive response were 0. Compared with excipient control group ，there was no statistical significance of average score and the incidence of hypersensitive response in test preparation group and commercial drug group. CONCLU- SIONS：In skin irritation test of rabbits and skin sensitization test of guinea pigs ， the evaluation results of generic Clobetasone butyrate cream are the same as those of the original drug. It has no irritation to the skin of rabbit ，and no sensitization to the skin of guinea pigs.

Objective To design bundle treatment plan in the early stage for severe human infection by avian influenza H7N9, and explore its clinical efficacy and application value. Methods Fifteen patients with severe human infection by avian influenza H7N9 in Guizhou Province from December 29th, 2016 to June 7th, 2017 were enrolled. Patients admitted from March 6th, 2017 to June 7th, 2017 served as a prospective observation period (bundle treatment group), and those from December 29th, 2016 to March 5th, 2017 were selected as a historical control period (conventional treatment group). Conventional treatment group was given conventional treatment such as isolation, anti-virus, symptomatic treatment, and traditional Chinese medicine and so on. Bundle treatment group was given bundle treatment on the basis of conventional treatment, including isolation, anti-virus, respiratory support, restrictive fluid management, immunotherapy, inhibition of inflammation, antibiotic therapy, nutritional support, prevention of hospital acquired infection (HAP), individual sedation, continuous blood purification (CBP) for acute kidney injury (AKI) and severe acute respiratory distress syndrome (ARDS) patients, and intensive care. A cluster of bundle treatment team was set up to ensure that all measures carried out smoothly. The gender, age, onset to diagnosis time, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score, oxygenation index (PaO2/FiO2) at admission, the length of intensive care unit (ICU) stay, total hospitalization time and prognosis of the two groups were observed. Correlation analysis between bundle therapy and prognosis was analyzed by Spearman correlation analysis. Receiver operating characteristic (ROC) curve was drawn, and the clinical value of bundle treatment was analyzed. Results There was no significant difference in gender, age, onset to diagnosis time, APACHEⅡscore, PaO2/FiO2, the length of ICU stay, or total hospitalization time between bundle treatment group (n = 9) and conventional treatment group (n = 6), but the death patients in the bundle treatment group was significantly fewer than those in conventional treatment group (cases:2 vs. 5, χ2= 3.225, P = 0.041). Correlation analysis showed that there was a significant correlation between the mortality and whether received bundle treatment or not in patients who infected by avian influenza H7N9 (r = -0.875, P = 0.018). ROC curve analysis showed that the area under the ROC curve (AUC) of non-bundle treatment for predicting the death in patients with severe human infection by avian influenza H7N9 was 0.938, 95% confidence interval (95%CI) was 0.795-1.000, the sensitivity was 88.88%, and the specificity was 98.62%. Conclusions Early bundle therapy has a significant effect on severe human infection by avian influenza H7N9, which can improve the prognosis and reduce the mortality of patients. It is worthy for clinical application.

A total of 108 subjects were enrolled in this study, including 21 healthy subjects(control group), 34 non-obese patients with polycystic ovary syndrome [PCOS1 group, body mass index(BMI)<25 kg/m²], 32 obese patients with PCOS(PCOS2 group, BMI≥25 kg/m²), and 21 simple obese patients whose age and BMI matched with PCOS2(OB group). BMI, waist-hip ratio(WHR), fasting plasma glucose(FPG), postprandial 2h plasma glucose(2hPG), HbA_1C, fasting insulin(FINS), postprandial 2h insulin(2hINS), sex hormones, and lipid parameters were determined. The status of insulin resistance was assessed by homeostasis model assessment for insulin resistant index(HOMA-IR)and insulin sensitivity index(ISI). Levels of plasma galectin-3 and interleukin-6(IL-6)were detected by ELISA. The results showed that the plasma level of galectin-3 was significantly higher in PCOS group than those in control and OB groups(all P<0.05). Moreover, the level of galectin-3 was also higher in OB group compared with control group, while galectin-3 level in PCOS2 group was higher than that in PCOS1 group(all P<0.05). After adjusted by age, BMI, and WHR, correlation analysis showed that the level of galectin-3 was positively correlated with FPG, 2hPG, FINS, HOMA-IR, highly-sensitive C-reactive protein, and IL-6, while negatively correlated with ISI. A multiple linear regression analysis revealed that the plasma galectin-3 concentration was independently associated with IL-6, HOMA-IR, and BMI(all P<0.05). These data suggest that plasma galectin-3 is closely associated with glucose metabolism, chronic inflammation, and insulin resistance, which may be involved in the pathogenesis of PCOS. (Chin J Endocrinol Metab, 2018, 34: 578-582)

Objective To observe the levels of neutrophil CD64 expression in prostatic fluid of patients with chronic prostatitis and different degrees of infection,to evaluate the value of CD64 in diagnosis and treatment of chronic bacterial prostatitis.Methods 100 subjects were divided into two groups according to chronic prostatitis symptom index (CPSI) established by the National Institutes of Health (NIH):CPSI high score group and CPSI low group,bacterial infection group,and 30 healthy controls.CD64 expression of neutrophils in prostatic fluid was detected by flow cytometry and prostate fluid routine microscopy inspection.Results The expression of CD64 in severe group (high score) was (4857.25±278.45) molecules / cell,which was significantly higher than that in the mild group (P＜0.05).There was significant difference between the mild group and the nonbacterial infection group or the healthy control group (P＜0.01).There was no significant difference between nonbacterial infection group and healthy control group.The expression of CD64 was positively correlated with some white blood cells in the bacterial infection group,but not the whole.ROC curve was drawn according to the data of the subjects.When CD64 was 2135.2 molecules/cell,the sensitivity was 92.9％ and the specificity was 91.7％.Conclusion Quantitative detection of CD64 in prostatic fluid can be used as a sensitive index for diagnosis of chronic bacterial prostatitis and has clinical value.

OBJECTIVE:To investigate the effects of different operation methods on the residual amount of docetaxel. METH-ODS:60 Docetaxel injections were selected and divided into two groups,i.e. standard operation method was adopted in group A (30 injections),and clinical common operation method was used in group B(30 injections). Docetaxel injections were placed for 5,10 or 15 minutes respectively,and drained. Residual liquid was collected to calculate residual amount. The difference of residual amount,due to preparation method and standing time,were compared between two groups. RESULTS:After placing for 5,10 or 15 min,the volume and amount of residual liquid in group B were significantly higher than in group A(P<0.05). In same group, the volume and amount of residual liquid after placing for 10 and 15 min were all lower than that after placing for 5 min(P<0.05 or P<0.01);the volume and amount of residual liquid after placing for 15 min were lower than that after placing for 10 min(P<0.05). CONCLUSIONS:The standard operation of pharmacy dispensing staff is very important to reduce the residual amount of docetaxel,and guarantee safe,effective and sufficient medication. It is suggested that clinical staff should abide by standard opera-tion method.

AIM:Early calcification of atherosclerotic plaques are colocalized with macrophage and high mobility group box 1 (HMGB1), a cytokine associated with biomineralizing process under physiological and pathological conditions .Our study aims to evaluate whether HMGB1 induces ectopic mineralization via promoting the secretion of matrix vesicles ( MVs) from macrophages .METHODS:HMGB1 was added to the medium of macrophages , the secretion of MVs in the supernatant was tested by flow cytometry analysis .The mineral deposition in calcifying medium was detected by Alizarin Red staining and von Kossa staining .Transmission electron microscopy showed the formation of hydroxyapatite crystals in MVs .Then we subcutaneous injection into mice with MVs to induce regional minera-lization.RESULTS:HMGB1 significantly promoted secretion of MVs from macrophages as raveled by flow cytometry analysis .TNAP activity, considered as a marker of MVs maturation , was higher in HMGB1-induced MVs compared to the control-MVs.HMGB1-MVs also led to mineral deposition in an in vitro MVs-collagen mineralization model .Subcutaneous injection into mice with MVs derived from HMGB1-treated cells showed a greater potential to initiate regional mineralization .Mechanistic experiments revealed that HMGB 1 activated neutral sphingomyelinase 2 ( nSMase2 ) that involved the receptor for advanced glycation end products ( RAGE ) and p38 MAPK (upstream of nSMase2).Inhibition of nSMase2 with GW4869 or p38 MAPK with SB-239063 prevented MVs secretion and min-eral deposition .CONCLUSIONS: HMGB1 induces MVs secretion from macrophages at least in part , via the RAGE/p38 MAPK/nSMase2 signaling pathway .Our findings thus reveal a novel mechanism by which HMGB 1 may participated in the early calcification of atherosclerotic plaques .

Objective To explore the improving effect of L-leucine on memory impairment in plateau and the mecha-nism. Methods After successfully trained in the 8-arm radial maze,50 male Kunming mice were selected and randomly divid-ed into normoxic control group (NC group),model group,and L-leucine (low,medium and high dose) groups.Animals in L-leu-cine groups were intragastrically given 0.473 g?kg-1 ,0.945 g?kg-1 and 1.89 g?kg-1 L-leucine for 7 days and those in NC and model control groups were administered the same volume of purified water for the same period of time.At the 4th day of the treat-ment,the mice in the model control group and L-leucine groups were placed in a large low-pressure and low-oxygen chamber to simulate low-pressure hypoxic environment of the plateau (7 500 m,3 d).The 8-arm radial maze was used to measure the spatial learning and memory ability of mice and dry-wet method to measure the water content of brain tissue.HE staining was employed to observe the cell morphological changes in CA1 region of the hippocampus.The expression levels of mTOR,P70S6K and 4E-BP1 mRNA in the hippocampus were detected by SYBR Green real-time PCR. Results The reference memory error ( RME) ,total error ( TE) ,testing time ( TT) ,and water content of brain tissue were significantly increased,the neuron injury was exacerbated in CA1 region of the hippocampus,and the expression levels of mTOR and P70S6K mRNA were markedly decreased in model control group when compared with those in NC group (P<0.05 or P<0.01).These indexes,however,were significantly improved in L-leu-cine groups,especially in high-dose group. Conclusion L-leucine can improve memory impairment in plateau,and the mecha-nism may involve the activation of mTOR and its downstream substrates (4E-BP1 and P70S6K).

Astract:Objective To investigate the ultrastructural changes and HSP70 expression in liver of mice after microwave irradiation with lethal dose and to explore the application of these indexes as the basis of medical identification in microwave irradiation induced death. Methods The mice were divided into the control group and the irradiation group. Mice of the irradiation group were induced death by whole body exposure to 129 W/cm2 microwave irradiation for 30 minutes. The ultrastructure of liver was observed by transmission electron micro-scope;changes of the HSP70 mRNA and protein expression in liver were detected by reverse transcription polymerase chain reaction ( RT-PCR) and Western blotting respectively. Results Liver cytoplasm was observed dissolved with points and sheets and there were mitochondri-al crest and membrane solution in the irradiation group. And the HSP70 mRNA and protein expression level increases significantly compared with the control group with statistically significant difference (P<0. 01). Conclusion Death induced by microwave irradiation could lead to liver cytoplasm dissolution, mitochondria damage, mRNA and protein expression of HSP70 up-regulation, which may be used as important diagnostic indicators of microwave irradiation induced death.