Objective To investigate the influence of HAp-CS composite microsphere scaffold on the in vitro cell behaviors of mesenchymal cells and evaluate its potential application for bone tissue engineering.Methods Nano-hydroxyap-atite (HAp)and chitosan (CS)composites solution were assembled into microsphere scaffold through microfluidic and observed by inverted microscope.Rat bone marrow mesenchymal stem cells were co-cultured in vitro with the microspheres for calculating the adhesion rate for the first 6h.Proliferation rate was measured by cell counting in the next 1,3,6,9 d,respectively,and GraphPad Prism 6 software was used for statistical analysis.The morphology of BMSCs on the surface of HAp-CS composite microsphere was observed by scanning electron micros-copy (SEM)and confocal scanning microscopy.The cells and HAp-CS microspheres were filled into a disc mold and co-cultured for 14 ~21 d to observe the morphology.Results HAp-CS microspheres were observed to be round and with uniform size by microscope.The adhesion rate of BMSCs reached 80% after cultured for 6 h,and proliferation rate reached the highest value when cultured for 6 d.SEM observations showed that BMSCs adhered compactly to the surface of the microspheres,and the microspheres could be connected together through BMSCs.Af-ter co-culturing BMSCs with microspheres for 14 ~21 d,a complete tissue constructs could be formed.Conclusion HAp-CS microspheres are proved to be good scaffolds for promoting BMSCs adhesion and proliferation.Large a-mount of extracellular matrix can be formed to connect microspheres after co-cultured for a certain time,which paves the way for HAp-CS microspheres to be applied for bone regeneration in animal experiments.

Objectives To investigate neuroprotective effect of melatonin on preterm rats brain damage after hypoxia-ischemia (HI). Methods In this study, 5-day-old Wistar rats were divided into four groups: normal saline group, melatonin group, HI+NS group and HI+melatonin group. HI was conducted by unilateral ligation of the left common carotid artery (ische-mia) and 50 min of hypoxia. Melatonin was injected at a dose of 5 mg/kg intraperitoneally three times:before ischemia, after hy-poxia and 24 h after the second dose. The pups were sacrificed at 24 h, 72 h, and 7 weeks after HI;for galectin-3 cells count at 72 h and 7 weeks;TNF-α, IL-1βprotein were measured in 24 h and 72 h after HI;and fear condition and elevated plus maze were tested in 7 weeks after HI. Results The number of galectin-3-positive cells was lower after melatonin treatment than vehi-cle treatment in 72 h and 7 weeks after HI (all P<0.05). TNF-αprotein and IL-1βprotein both increased at 24 h and 72 h after HI, and reduced after melatonin treatment (all P<0.05). Melatonin treatment improved memory ability and learning ability, re-duced anxiety in 7 weeks after HI. Conclusions Melatonin has long-term and short-term protective effect on developing brain damage after HI.

Objective To observe the effect of Gongxuening for the women placing the IUDafter cesarean section, and to analyze the effect and value of Gongxuening on intrauterine device placement after cesarean section. Methods 156 women who received voluntary intrauterine device after cesarean sectionin theShengzhou Maternal and Child Health Hospital of Shaoxing, Zhejiang were as the observation object. According to whether the application of Gongxuening, they were divided into two groupsafter treatment, the Gongxuening group of 75 cases (using Gongxuening + Nifloxacin) and the control group of 81 cases (using norfloxacin). The changes of serum inflammatory indexes were compared between the two groups before and after operation, and the uterine bleeding was compared between the two groups. Results After operation, the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) were with a significantfluctuation, and at the same time, Gongxuening group TNF-α, IL-6, IL-10, 24hours after surgery, postoperative 7d was significantly lower than the control group, the difference was statistically significant (P< 0.05). After treatment, the WBC of Gongxuening group 24hours after surgery, postoperative 7d was significantly lower than the control group, but the Hb 24 hours after surgey was significantly higher than the control group, the difference was statistically significant (P<0.05). The bleeding volume2h, 24h, bleeding stop time and puerperium duration in the Gongxuening group were significantlylower than the control group, the difference was statistically significant (P<0.05). Conclusion After cesarean section maternal IUD use Gongxuening capsule treatment, not only can effectively inhibit the expression of serum inflammatory markers, and can shorten the time of uterine bleeding, reduce bleeding, prevent postoperative anemia. It is suggested to pay more attention to clinical treatment.

Objective:To analyze the skin microbiota diversity in patients with pemphigus vulgaris (PV) using 16S rDNA sequencing.Methods:Ten patients with PV and 10 healthy controls were collected from the Department of Dermatology, the Third People′s Hospital of Hangzhou. Skin swabs were collected from perilesional skin (PV group) and contralateral non-lesional skin (PVn group) of the patients with PV, as well as from the corresponding body sites of the healthy controls (normal control group) . The 16S rDNA amplicon sequencing technology was used for gene sequencing and classification in all microbiota samples, and Usearch software for data cluster analysis to obtain operational taxonomic units (OTUs) and assess species abundance at the phylum, class, order, family and genus levels. Observed species index, Shannon index and Simpson index were used to estimate α diversity, and principal coordinate analysis (PCoA) was performed to analyze β diversity. Linear discriminant analysis effect size (LEfSe) analysis was conducted to identify differentially abundant species in each group. PICRUSt software was used for gene function prediction. Wilcoxon rank sum test was used as nonparametric test for comparisons between 2 groups, and Kruskal Waills test as nonparametric test for comparisons among 3 groups.Results:There were 2 002, 1 869, 1 751, 1 611 and 1 120 OTUs at phylum, class, order, family and genus levels respectively. Cluster analysis showed that skin microbiome in the 3 groups mainly consisted of Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria at the phylum level. At the genus level, Staphylococcus was the most abundant in the PV group and PVn group, and Corynebacterium was the most abundant in the normal control group. The observed species index, Shannon index and Simpson index all significantly differed among the 3 groups (all P< 0.05) , and the Shannon index and Simpson index were significantly lower in the PV group (3.24±1.30, 0.70±0.19, respectively) than in the normal control group ( P< 0.05) . PCoA analysis showed no significant difference in β diversity among the 3 groups ( P=0.054) . Rank sum test showed that the abundance of 32 species significantly differed among the 3 groups ( P< 0.05) . Among them, high relative abundance was observed in the class Bacilli enriched in the PV group, as well as the genera Micrococcus and Brevundimonas enriched in the normal control group. According to the disease duration, the patients with PV were divided into long-course PV group with disease duration of ≥ 3 months, and short-course PV group with disease duration of < 3 months. Clostridiales, Oscillibacter, Sphingomonas were enriched in the long-course PV group, and Gammaproteobacteria was enriched in the short-course PV group. Gene function prediction analysis showed that the genes related to infectious diseases were enriched in the pemphigus group. Conclusion:The 16S rDNA-based microbiota profiling suggested differences in the diversity and composition of skin microbiota between patients with PV and healthy individuals.

Objectives To study the influence of Staphylococcus epidermidis (SE) on the neonatal mice of different ages. Methods A total of 60 neonatal mice including postnatal day 1(PND1) and postnatal day 3(PND3) were divided into SE group, normal saline (NS) group and control group, with 20 mice each. Mice in SE group were intravenously injected with 50μl SE (108/ml). Mice in NS group were given 50μl NS and mice in control group was not intervened. On postnatal day 14, the brain, liver and spleen obtained from mice were weighted. Serial sections of paraffin-embedded brain tissue were used for the detec-tion of microtubule associated protein-2 (MAP-2) and myelin basic protein (MBP) by immumohistochemical staining, and then the areas and volumes of grey and white matter were calculated. Result The mortality of PND1 mice in SE and NS group was 60.0%and 40.0%, respectively, and there was no difference between two groups (P>0.05). The mortality of PND3 mice in SE and NS group was 10.0%and 0.0%, respectively, and there was no difference between two groups (P>0.05). There were no dif-ferences in body weight, body weight gain, spleen and liver weights and organ coefficient between PND1 and PND3 mice (P>0.05). In PND1 mice, the areas and volumes of grey and white matter were significantly smaller in SE group than those in NS group (P<0.05). However, in PND3 mice, there was no differences in areas and volumes of grey and white matter between SE and NS group (P>0.05). Conclusions SE infection can result in brain injury in PND1 mice, but has no effect on brain tissues of PND3 mice.

ObjectivesTo study the mechanism of brain damage caused byStaphylococcus epidermidis (SE) in mice. Methods A total of 80 neonatal mice of postnatal day 1 (PND1) were divided into SE group (48 mice), normal saline (NS) group (16 mice) and control group (16 mice). Mice in SE group were intravenously injected with 50 μl SE (108/ml). Mice in NS group were given 50 μl NS. Mice in control group were not intervened. At different time points after SE injection (6 h, 24 h, 72 h, 5 d, 7 d), the CFU of brain, blood, and spleen were calculated. Serial sections of parafifn-embedded brain tissue were used for detection of ionized calcium-binding adaptor moleculor1 (Iba-1) by immunohistochemical staining. The positive cells were calculated. ELISA was used to measure the levels of tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), interleukin-5 (IL-5), interleukin-6 (IL-6) of brain at 6 h and 24 h after SE injection.Results There was no SE in brain in different time points. The CFU was at the highest level at 6 h and then decreased after 24 h in blood and spleen. The Iba-1 positive cells in SE group were signiifcantly increased compared to NS group and control group at 24 h and 72 h (P<0.05). There was no difference of Iba-1 positive cells be-tween 24 h and 72 h after SE injection (P>0.05). The levels of TNF-α, IL-1β, IL-5, and IL-6 were signiifcantly higher in SE group than those in NS and control at 6 h and 24 h (P<0.05). The levels of TNF-α, IL-1β, IL-5, and IL-6 were signiifcantly lower in SE group at 24 h than those in SE group at 6 h (P<0.01).Conclusions It is suggested that cytokines produced by microglias may be the mediators of SE-caused brain damage.

Objective To research early diagnosis and treatment of acute spontaneous spinal epidural hematoma (SSEH). Methods Medical records of 13 SSEH patients adimtted in Timone Ste Marguerite Hospital,France and Ruijin Hospital,China from 1985 to 2000 were retrospectively reviewed.The etiology, neurological symptoms, neuroradiology therapy, as well as the prognosis of this rare disease are discussed in comparison with the literaturs.Results Six of thirteen SSEH wee relatd with innormal coagulation. All patients had the same original symptom which was radicular pain. Eleven cases were diagnosed by MRI. After decompressive surgery, recovery occurred in 3 of 5 patients. Five of 8 patients had a favorable outcome after medical treatment. Conclusions The majority of spontaneous hematomas results from a rupture of the venous plexus. It could be diagnosed according typical symptoms and MRI. Decompressive surgery is urgent but not unique. A idiosyncratic type which has a spontaneous complete recovery could be found.

OBJECTIVE:To discuss the effect of tanshinone Ⅱ A on the apoptosis of HeLa cell line and its mechanism.METHODS:MTT was used to measure the levels of the proliferation of HeLa cells cultured with tanshinone Ⅱ A in different concentrations.The apoptosis was measured by Hoechst33258 staining,and the level of mRNA of Bcl-2 and bax in HeLa cells was tested by semi-fix quantify RT-PCR method.RESULTS:Tanshinone Ⅱ A could inhibit the proliferation of HeLa cells in a dose dependent manner,induce cell apoptosis,and markedly decrease the expression of Bcl-2 on mRNA level after treatment for 48h,while it had no obvious change in bax mRNA level.CONCLUSION:Tanshinone Ⅱ A can inhibit the proliferation of HeLa cells and induce the apoptosis of the cells.

OBJECTIVE:To outline the development of phytomedicine in Germany to offer a reference for traditional Chinese medicine.METHODS:From the aspects of research,production and market to make an exposition of phytomedicine's develop?ment in Germany.RESULTS:There existed a gap between Germany and China in technology and standards in production of botanicals.CONCLUSION:We should use the experience of Germany in developing botanicals to draw up international quality standard of traditional Chinese medicine.

Objective Aspirin is a non steroidal anti-inflammatory drugs, whether or not which does harm to renal function in the course of using, there is a big debate at home and abroad.In this paper, The aim of this study was to observe the effects of different doses of aspirin on renal function in elderly patients with cardiovascular disease at different ages.MethodsWe Selected a total of 288 patients with cardiovascular disease from January 2014 to March 2016 in Zhejiang Rongjun hospital.There were 288 cases of cardiovascular disease, including 144 cases of normal uric acid, 144 cases with hyperuricemia (including gout).The two groups according to the age of 60 to 70 years, 71 to 80 years old, over 80 years old respectively into three subgroups, each subgroup of 48 cases;Each subgroup was randomly selected without the use of aspirin (0mg/d) and the use of different doses of aspirin (50mg/d, 100mg/d, 150mg/d) for a total of four groups, each which is 12 patients.We detect patients routine blood tests, liver and kidney function, blood urea nitrogen (BUN), blood creatinine (SCR) levels and glomerular filtration rate (GFR) (MDRD) and blood and urine beta 2-microglobulin used aspirin before and used after 6 months and 12 months.ResultsBefore treatment, there were no significant differences in body mass index (BMI), blood pressure, hemoglobin, serum albumin and platelet count between various groups of the same age group.After treatment for 6 and 12 months,Uric acid normal patients and high uric acid patients with 0mg/d, 50mg/d, 100mg/d or 150mg/d dose of aspirin has no statistical difference in BUN, Scr and eGFR, 2-microglobulin Ccr, blood, urine beta 2-microglobulin.ConclusionRegardless of uric acid, different doses of aspirin had no significant effect on renal function in elderly patients with cardiovascular disease, and can be safely used.