Objective To develop a matrix metalloproteinase(MMP)-responsive hyaluronic acid(HA)-based controlled-release material for brain-derived neurotrophic factor(BDNF)to provide a novel therapeutic strategy for intervention and repair of traumatic brain injury(TBI).Methods HA was modified with amination,followed by condensation with Suflo-SMCC carboxyl group to form amide,and then linked with glutathione(GSH)to synthesize HA-GSH.The recombinant glutathione S-transferase(GST)-tissue inhibitor of metalloproteinase(TIMP)-BDNF(GST-TIMP-BDNF)expression plasmid was constructed using molecular cloning technique with double enzyme digestion by Bam H Ⅰ and Eco R Ⅰ.The recombinant GST-TIMP-BDNF protein was expressed in the Escherichia coli prokaryotic expression system,and purified by ion exchange chromatography,confirmed by Western blotting.MMP diluents were supplemented with PBS,MMP inhibitor marimastat,and varing concentrations(0.4,0.6,0.8 mg/ml)of GST-TIMP-BDNF or GST-BDNF.MMP-2 activity was analyzed using an MMP activity detection kit to evaluate the inhibitory effect of the recombinant protein on MMP.Primary rat neurons were extracted and cultured to establish an iron death model induced by RSL3.The effect of recombinant protein GST-TIMP-BDNF on neuronal injury was detected by immunofluorescence staining.Results MRI hydrogen spectrum identification confirmed the successful synthesis of HA-GSH.Western blotting results showed the successful expression of the recombinant protein GST-TIMP-BDNF containing the GST tag using the E.coli prokaryotic expression system.MMP activity detection results indicated that the recombinant protein GST-TIMP-BDNF had a superior inhibitory effect on MMP-2 activity compared to GST-BDNF(P<0.05).Immunofluorescence staining results showed a significant increase in fluorescence intensity in rat neurons treated with GST-TIMP-BDNF after RSL3 induction(P<0.05).Conclusion A MMP-responsive HA-based BDNF controlled-release material has been successfully developed,exhibiting a protective effect on neuron damage.

Objective: Cimicifuga dahurica (Ranunculaceae) has many bioactivities. Although there have been intensive studies for saponin constituents at present, only a few studies have focused on for chemical constituents of phenolic acid. To define the phenolic acid constituents,C. dahurica was separated, and the structures of the compounds were identified,in the expectation of providing a basis for its further development,utilization and quality control. Method: A total of 16 kg rhizome of C. dahurica was extracted with 70%ethanol for three times by heating reflux. These 3 extracts were decompressed and concentrated,and then dissolved in water. Then the solvent was successively extracted with petroleum ether,ethyl acetate(EtOAc) and n-butanol(BuOH). The components of EtOAc and water extract were isolated and purified by macroporous,silica gel,ODS,Sephadex LH-20 column chromatography,preparative HPLC and recrystallization,and the structures were identified by nuclear magnetic resonance(NMR) and physicochemical analysis etc. Result: Fifteen compounds were isolated from the ethyl acetate and water fractions,and identified as cimicifugic G (1),2-caffeoyl piscidic acid (2),cimicifugic A (3),cimicifugic B (4),caffeic acid 3-O-β-D-glucopyranoside (5),cimicifugic E (6),cimicifugic F (7),trans-ferulic acid 4-O-β-D-glucopyranoside (8),carboxymethyl isoferulate (9),3,4-dimethoxycinnamic acid (10),ethyl ferulate (11),caffeic ester glucoside (12),shomaside A (13),isoferulic acid (14),caffeic acid (15). Conclusion: Compounds 1-7,9-10,13 were isolated from the plant for the first time.

Paecilomyces hepiali is a new species of fungus isolated from a field collection of Ophiocordyceps sinensis from Baima snow mountain, Diqing Tibetan Autonomous Prefecture, Yunnan Province by the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences. The specimen was identified and named as Paecilomyces hepiali by Qing-Tao Chen, the professor of the Institute of Microbiology, Chinese Academy of Sciences (Paecilomyces hepiali) (2008), who identified a dried culture of living strain 82-2 as the holotype. Until now, the holotype (the voucher specimen) was deposited in the Herbarium of the Institute of Chinese Materia Medica (HICMM), China Academy of Chinese Medical Sciences, Beijing. The P. hepiali neotype designated by the paper "Neotypification of P. hepiali (Hypocreales)" published in TAXON 64 (1) by Yao Yi-Jian et al. in February 2015 is untenable.

OBJECTIVETo investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs.

METHODBeagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration.

RESULTCompared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine.

CONCLUSIONMethyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Animals ; Biomarkers ; urine ; Dogs ; Female ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Male ; Proteins ; metabolism ; Tablets ; adverse effects ; Urine ; chemistry

BACKGROUNDThe sensitization and elicitation phases are involved in the immunopathogenesis of contact hypersensitivity (CHS). Langerhans cells (LCs) are believed to play pivotal roles in the sensitization stage of CHS. Local hyperthermia on skin induces the migration as well as maturation of epidermal LCs. Although fever-range whole body hyperthermia and local hyperthermia at 43°C prior to sensitization were reported to suppress CHS, the effects of different temperatures and the timing sequence of local hyperthermia on CHS have not been tackled.

METHODSLocal hyperthermia was applied to murine dorsal skin 3 days prior to, concurrent with, or 2 days post sensitization with fluorescein isothiocyanate (FITC) in BALB/c mice. Local hyperthermia temperatures at 37°C, 39°C, 41°C and 43°C were applied to mouse dorsal skin and the severity of CHS was calculated by measuring the swelling response of the challenged ears.

RESULTSLocal hyperthermia at 39°C, 41°C and 43°C prior to sensitization reduced the severity of CHS, as compared with that at 37°C. The suppression of CHS was temperature dependent in that higher temperature had a stronger effect. On the contrary, the hyperthermia treatments, either concurrent with or post-sensitization, resulted in an enhanced temperature-dependent ear swelling response.

CONCLUSIONSThe severity of murine CHS could be influenced by local hyperthermia at the sensitization stage in a temperature dependent manner. The temporal effect of local hyperthermia suggested a novel factor in interpreting the severity of allergic contact dermatitis.

Animals ; Dermatitis, Contact ; therapy ; Female ; Hyperthermia, Induced ; Langerhans Cells ; physiology ; Mice ; Mice, Inbred BALB C

Objective To study the clinical significance of the method of three perpendicular planes plus special planes in diagnosing fetal cleft lip /palate by prenatal ultrasound .Methods The approach of three perpendicular planes and special planes were used in diagnosing 110 cases of cleft lip/palate.The sonogram features in each section were analyzed and the outcomes were recorded during follow-up.Results On prenatal ultrsound ,110 cases were examined with three perpendicular planes method .The coronary section could be displayed at 100%cases (110 cases), sagittal section 76.4%cases (84 cases),transverse section 96.4%cases (106 cases) and parasagittal section 25.5%cases (28 cases).With special planes method,74 cases were examined .The section through pyriform aperture could be displayed in 47 cases,in 45 cases on the section through the lower lip/lower jaw/submandibular triangle ,and in 16 cases on the section through the cheek.Combining the three perpendicular planes and special planes methods ,94.5%(104/110) cases could be diagnosed definitely.Six cases (5.5%,6/110) were missed because of fetal position or oligoamnios . Conclusions The method of three perpendicular planes plus special planes is effective in prenatal ultrasound diagnosing cleft lip/palate,which is of great help in improving prenatal diagnostic accuracy of fetal cleft lip/palate.

Objective To investigate the effects of enamel matrix proteins (EMP) on odontogenic differentiation of human dental pulp cells (HDPC) in vitro.Methods HDPC were isolated by explant culture methods and expanded to passage 4 for experiments.The cells were seeded into 6-well plates (2 × 105 cells/well) and divided into 5 groups,treated with 1,10 or 100 mg/L EMP,10-8 mol/L dexamethasone and 100 mg/L ascorbic acid (Dex-AA) or basal media respectively.Alkaline phosphatase (ALP) activity was measureded after up to 10 days in culture.The mRNA expression of dentin matrix protein-1 (DMP-1)and dentin sialophosphoprotein (DSPP) were determined by quantitative real-time reverse transcriptase polymerase chain reaction.Calcified nodule formation was evaluated by alizarin red staining and spectroscopic method.Results The ALP activity levels of all 5 groups were increased in a time-dependent manner.There was no significant difference among each group after 1 day.After 5 days,the ALP activity was increased significantly especially for the 10 mg/L EMP,100 mg/L EMP and Dex-AA groups,values were 7.573 ± 0.267,6.119 ± 0.502 and 5.846 ± 0.096,respectively (P ＜ 0.05).After 10 days,the increase of ALP activity in the 10 mg/L EMP(21.035 ± 0.149) and Dex-AA groups(13.223 ± 0.797) was remarkable,compared with the negative control group (5.825 ± 0.404) (P ＜ 0.01).Statistical differences were detected for DMP-1 and DSPP mRNA expressions in 10mg/L EMP group(14.791 ± 0.164,12.238 ± 0.421),compared with negative control group (4.959 ± 0.184,2.645 ± 0.570) (P ＜ 0.01).Phase contrast microscopy revealed orange-red stained nodules in 1 mg/L and 10 mg/L EMP groups.Such staining was not that much obvoius in the negative control group.After quantifying the staining,it can be clearly seen that 10 mg/L EMP enhanced calcium deposition [(191.8 ± 2.0) μmol/L] significantly,compared to the negative control[(81.1 ± 8.1) μmol/L] (P ＜ 0.01).Conclusions This study showed that optimized concentration of EMP can stimulate odontoblastic differentiation of HDPC,which suggests that EMP may be useful for directing HDPC down the odontoblastic lineage in dentine tissue engineering.

Objective To investigate the prevalence of unintended pregnancy (UP) and exploring the risk factors of UP for married women of child-bearing age from Qingshan district,Wuhan.Methods A cross-sectional study was adopted in this study.Cluster sampling method was used with 3256 women recruited,in 2010.Information on history and risks related to social-demographic factors of UP were collected,using a self-administered questionnaire.Results Of the 3256 participants,over half of them (53.8％) reorted ever having had the history of UP and 9.1％ reported UP in the past year.Rate of UP in the past year for different age cohorts (18-30,31-40,41-49 years) were 31.8％,10.5％ and 1.8％ respectively.The most frequently reported reason for UP across all the age cohorts was "Did not use any contraceptive methods",with proportions on the reason that reported by women at 18-30,31-40 and 41-49 year-olds,were 69.7％,51.1％ and 42.4％ respectively.The second frequently reported reasons for UP were "Failure of traditional contraception" for younger cohort ( 18-30 years:13.0％ ) and "IUD dropped or pregnancy with IUD" for older-age cohorts (23.4％ at 31-40 year-olds and 37.0％ at the 41-49 year-oplds).The most frequently cited reason for "Did not use any contraceptive methods" was "Believe we were lucky so far,not to get pregnant" (59.6％).The risk factors of UP were being at older age,experiencing sex debut at younger age and got married at younger age.Conclusion The prevalence of lifetime UP history was high among women at child-bearing age from Qingshan district,Wuhan.Reproductive health services and interventions should be taken according to the needs from different age cohorts of women.Younger cohort of women should receive more attention.

Objective To investigate the effect of chewing gum after cesarean section on restoration of gastrointestinal function.Methods 300 patients who had undergone low cesarean section in our hosplta|were randomlv divided into observation group(150 cases)and control group(150 eases).The former received sugarfree chewing gum 4 times per day(2～4 pieces each time)from 2h to 3d after cesarean section and the latter received routine nursing care. Gastrointestinal function of the two groups Was observed.Results Earlier alleviation of abdominal distension.as well as resume of bowelsound，anal exhaust and defecation was found in the observation group compared to the control group(P<0.05).Conclusions Chewing gum can promote restoration of gastrointestinal function and thus benefits rehabilitation of patients after cesarean section.

BACKGROUNDImplantation of tissue-engineered scaffolds is one of the most promising therapeutic strategies for inducing nerve regenerations following spinal cord injuries. In this paper, we report a novel bioengineered hybrid scaffold comprised of three major extracellular matrix (ECM) proteins.

METHODSECM-scaffolds (ECM-S) were prepared by gelling fibrinogen, fibronectin and laminin using fresh rat plasma. Olfactory ensheathing cells (OECs) were isolated from fresh rat olfactory mucosa, purified under differential adhesion, and assessed by immunofluorescent staining. OECs were seeded onto ECM-S and cultured. The effects of the scaffolds on the seeded cells were detected using the immunofluorescent staining, Western blotting, scanning electron microscopy and transmission electron microscopy.

RESULTSTissue-engineered ECM-S could be easily molded into mat-like or cylindrical shapes and gelled by addition of fresh plasma. Observations by electron microscopy show that the ECM-S forms a stable three-dimensional porous network. Studies on the effects of the ECM-S on the biological behaviors of OECs in vitro indicate that the scaffold can promote OEC adhesion, proliferation and process extensions. Additionally, OECs seeded on the scaffold maintained the expression of nerve growth factor, matrix metalloproteinase-3 and matrix metalloproteinase-9.

CONCLUSIONWe developed a biosynthetic hybrid gel which could be used as a scaffold for OEC transplantation; this gel can promote nerve regeneration following spinal cord injuries.

Animals ; Cells, Cultured ; Immunoblotting ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Olfactory Bulb ; cytology ; physiology ; transplantation ; Olfactory Mucosa ; cytology ; physiology ; transplantation ; Rats ; Spinal Cord Injuries ; therapy ; Tissue Engineering ; methods ; Tissue Scaffolds