Objective: To observe the effectiveness and safety of electroacupuncture (EA) plus Luohua Anshen oral liquid for patients with perimenopausal insomnia. Methods: A total of 66 participants who met the inclusion criteria were enrolled in the randomized controlled trial and allocated to a treatment group and a control group at a ratio of 1:1, with 33 cases in each group. Both groups were given Luohua Anshen oral liquid as a basic treatment. The treatment group was additionally given EA every other day, three times a week. Both groups were treated for four weeks and a four-week follow-up was conducted. The scores of Pittsburgh sleep quality index (PSQI), Kupperman index (KI) and traditional Chinese medicine sleep syndrome scale (TCMSSS) were recorded at pre- and post-treatment, and at the follow-up. Meanwhile, adverse effects were monitored and recorded. Results: After four-week treatment, the global scores of PSQI, KI and TCMSSS in both groups declined significantly (all P<0.05), and the decreases in the treatment group were more significant than those in the control group (allP<0.05). The global scores of PSQI, KI and TCMSSS in both groups at the follow-up visit were significantly different from the corresponding baseline (allP<0.05), while insignificantly different from those assessed at post-treatment (allP>0.05). The total effective rate was 93.9％ in the treatment group, significantly higher than 72.2％ in the control group (P<0.05). No significant adverse event was reported in this trial excepted one patient experienced slight dizziness in the first acupuncture treatment. Conclusion: EA plus Luohua Anshen oral liquid is safe for perimenopausal insomnia with satisfactory short- and long-term effectiveness, and it shows certain advantage compared with using Luohua Anshen oral liquid alone.

Objective To explore the technique of maximum entropy model for extracting Oncomelania hupensis snail habi?tats in Poyang Lake zone. Methods The information of snail habitats and related environment factors collected in Poyang Lake zone were integrated to set up the maximum entropy based species model and generate snail habitats distribution map. Two Land?sat 7 ETM+remote sensing images of both wet and drought seasons in Poyang Lake zone were obtained,where the two indices of modified normalized difference water index(MNDWI)and normalized difference vegetation index(NDVI)were applied to ex?tract snail habitats. The ROC curve,sensitivities and specificities were applied to assess their results. Furthermore,the impor?tance of the variables for snail habitats was analyzed by using Jackknife approach. Results The evaluation results showed that the area under receiver operating characteristic curve(AUC)of testing data by the remote sensing?based method was only 0.56, and the sensitivity and specificity were 0.23 and 0.89 respectively. Nevertheless,those indices above?mentioned of maximum en?tropy model were 0.876,0.89 and 0.74 respectively. The main concentration of snail habitats in Poyang Lake zone covered the northeast part of Yongxiu County,northwest of Yugan County,southwest of Poyang County and middle of Xinjian County,and the elevation was the most important environment variable affecting the distribution of snails,and the next was land surface tem?perature(LST). Conclusions The maximum entropy model is more reliable and accurate than the remote sensing?based meth?od for the sake of extracting snail habitats,which has certain guiding significance for the relevant departments to carry out mea?sures to prevent and control high?risk snail habitats.

Objective:To observe the efficacy and safety of dexzopiclone plus auricular acupressure in intervening primary insomnia.Methods:A total of 72 participants who met the inclusion criteria were enrolled in a randomized controlled trial,with 36 cases allocated to a treatment group and 36 cases allocated to a control group.Both groups were given dexzopiclone as the routine treatment.Patients in the treatment group were given auricular acupressure with Wang Bu Liu Xing (Semen Vaccariae) seeds at the auricular acupoints related to sleep and emotion based on meridian theory,whereas for patients in the control group,the medical plasters with Wang Bu Liu Xing (Semen Vaccariae) seeds were only gently stuck to acupoints unrelated to sleep without stimulation.Patients in both groups were required to visit the hospital once a week for replacing the seeds and plasters.The course of intervention lasted for 8 weeks and the patients were followed up for another 4 weeks.Pittsburgh sleep quality index (PSQI) and Karolinska sleep diary (KSD) were used to evaluate the outcomes.Meanwhile,adverse effects were monitored and recorded.Results:In the enrolled 72 cases,4 patients (one in the treatment group and three in the control group) reported thirst and a bitter taste,and one case in the control group reported nausea and vomiting.At last,3 cases in the control group dropped out for adverse reactions,and 69 cases completed the clinical trial.After 8 weeks of treatment,the global scores of PSQI in both treatment and control groups decreased significantly compared with the baseline (both P＜0.001).Furthermore,the global score of PSQI in the treatment group was lower than that in the control group (P＜0.01).The global scores of PSQI in both groups at the follow-up were significantly different from the baseline (both P＜0.001),but insignificantly different compared with the post-treatment results (both P＞0.05).According to KSD,both treatment protocols could prolong the total sleep time,shorten sleep-onset latency,improve sleep efficacy and sleep quality significantly,and the changes in the treatment group were more significant.The total effective rate was 88.9％ in the treatment group,higher than 81.8％ in the control group,though the difference was statistically insignificant (P＞0.05).Conclusion:Dexzopiclone plus auricular acupressure is effective and safe for patients with primary insomnia both in short and long terms,and it is more effective than monotherapy of dexzopiclone.

OBJECTIVETo understand the etiology of hand, foot and mouth disease (HFMD) in Guangzhou area in 2008.

METHODTotally 1023 clinical specimens were collected from pediatric patients suspected of HFMD in 2008. TaqMan real-time RT-PCR were used for detection of enterovirus 71 (EV71), Coxsackievirus A16 (CA16) and other enteroviruses. The specimens which were enterovirus positive by RT-PCR method with universal primer but EV71 and CA16 negative, were amplified and sequenced for 5'untranslated region.

RESULTEnterovirus was identified from 434 of 1023 samples and detection rate of enterovirus was 42.42%; of the 434 samples, 276 were positive for EV71 (63.6%), 126 for CA16 (29%), 4 samples for enterovirus 84, 3 for Echovirus 11, 2 for Echovirus 9, 3 for Coxsackievirus B3, 4 for Coxsackievirus A10, 3 for Coxsackievirus A6, 6 for Coxsackievirus A12 or A5, and for 7 samples typing was difficult.

CONCLUSIONThe major causative agents of HFMD in Guangzhou were EV71 and CA16 in 2008, and EV84, CA10, CA12, CA6, COSB3, ECHV11, ECHV9 were also the pathogens for smaller proportions of patients.

Child ; Child, Preschool ; China ; epidemiology ; Coxsackievirus Infections ; epidemiology ; DNA Primers ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; virology ; Humans ; Infant ; Male ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo find out a possible approach to improve the effectiveness of radiotherapy and chemotherapy for Ewing's sarcoma by constructing a eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia and to evaluate the effects of this HRE regulated HSV-TK system on killing effect of gancyclovir (GCV) on Ewing's sarcoma cell line SK-ES under hypoxic condition.

METHODSThe HRE was synthesized according to the literature and cloned into the enhancer site of pIRES(2)-EGFP vector to obtain the pHRE recombinant plasmid. The HSV-TK was amplified by PCR and cloned into the multiple clone site of pIRES(2)-EGFP and pHRE to obtain pTK and pHRE-TK recombinant plasmid. The human Ewing's sarcoma cell line SK-ES was transfected by pTK or pHRE-TK recombinant plasmid with liposome and then was exposed to normoxic (21% oxygen) or hypoxic (3% oxygen) condition. The expression of enhanced green fluorescent protein (EGFP) was monitored by fluorescent microscopy. The sensitivity of human Ewing's sarcoma cell line SK-ES transfected with pTK or pHRE-TK recombinant plasmid to the anti-tumour drug GCV was determined with the method of tetrazolium (MTT) after treating with GCV for five days.

RESULTS(1) The result of sequencing showed that the recombinant plasmid pHRE contained HRE, and that the recombinant plasmid pTK and pHRE-TK contained HSV-TK gene in the sense direction. (2) Comparison of fluorescent optical density (FOD) showed that (1) the EGFP FOD value of pHRE and pHRE-TK group cells exposed to hypoxia was significantly higher than those exposed to normoxia (P < 0.01); (2) when the cells were exposed to hypoxia, the EGFP FOD value of pHRE and pHRE-TK group cells was significantly higher than that of pTK and empty vector group (P < 0.01); (3) there was no significant difference among the four groups of cells when they were exposed to normoxia (P > 0.05). (3) Comparison of the sensitivity of four groups of cells to GCV showed that (1) the cells in pHRE-TK and pTK groups were much more sensitive to GCV than the cells in pHRE group under hypoxia condition (P < 0.01), the higher the GCV concentration, the greater the difference; (2) the cells of pHRE-TK group were more sensitive to GCV than those in pTK group under hypoxic condition (P < 0.01), but was almost equally sensitive under normoxic condition (P > 0.05); (3) the pHRE-TK group cells had higher sensitivity to GCV under hypoxia than normoxia (P < 0.01) while the pTK group cells had almost the same sensitivity to GCV under hypoxia and normoxia (P > 0.05).

CONCLUSION(1) The eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia was constructed successfully. (2) HRE could up-regulate expression of EGFP by SK-ES cells under hypoxia condition. (3) HRE could enhance the killing effect of HSV-TK/GCV system on human Ewing's sarcoma cell line SK-ES under hypoxic condition.

Antiviral Agents ; pharmacology ; Cell Hypoxia ; drug effects ; genetics ; Cell Line, Tumor ; Ganciclovir ; pharmacology ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Microscopy, Fluorescence ; Plasmids ; Polymerase Chain Reaction ; Response Elements ; genetics ; Sarcoma, Ewing ; drug therapy ; metabolism ; Simplexvirus ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection

Stimulated by retinoic acid gene 8 (STRA8) is specifically expressed in mammalian germ cells before their transition from mitosis to meiosis. STRA8 expression is observed only in the postnatal testis. Single nucleotide polymorphisms but no mutations were identified in the coding or proximal promoter region of STRA8 in some gonadal dysgenesis patients. Studies on the teratocarcinoma cells and embryonic stem cells (ESC) transfected with STRA8-EGFP, a fusion construct harboring the promoter and coding region of the enhanced green fluorescence protein, have shown that the STRA8-EGFP positive cells may undergo meiosis, develop into sperm and generate live offspring mice. STRA8-EGFP positive cells derived from the bone marrow are able to differentiate into spermatogenic cells, but arrest in the premeiotic stage, and those from the adult mouse testis, when cultured in ESC culture conditions, may acquire ESC properties, pluripotency and redifferentiation capacity and act as a new stem cell source for tissue regeneration. The presence of oocytes renewed in postnatal mouse ovaries calls in question the absence of STRA8 in postnatal mouse ovaries.
Adaptor Proteins, Signal Transducing ; Animals ; Biomarkers ; metabolism ; Cells, Cultured ; Female ; Germ Cells ; cytology ; metabolism ; Humans ; Male ; Mice ; Oocytes ; metabolism ; Proteins ; metabolism ; Testis ; cytology ; metabolism

OBJECTIVETo investigate the protective action of Epimedium against chemotherapy-induced damage to rat epididymides.

METHODSFifty 60-day-old male rats were divided into a control, a model and a treatment group. Procarbazine was injected into the abdominal cavity of the model rats at the dose of 30 mg/(kg x d). In addition to procarbazine, Epimedium was given intragastrically to the treatment group. The changes in the ultrastructure of the epididymis were observed after 10 and 20 days.

RESULTSElectron microscopy showed that the chemotherapy-induced damages to the epididymal epithelia mainly included cell swelling, local cavitation of mitochondria, tumor-like change in nucleoli, agglutination of marginal translocation of heterochromatin and cell apoptosis. The damage to the epithelial ultrastructure was slight in the treatment group as compared with the model rats. Chemotherapy significantly affected sperm concentration, sperm viability and sialic acid (SA), which were (15.59 +/- 4.01) x 10(6)/ml, (76.71 +/- 10.11)% and (19.38 +/- 9.34) g/mg prot in the model group in comparison with (10.63 +/- 3.82) x 10(6)/ml (P < 0.01), (60.03 +/- 7.54)% (P < 0.01) and (13.62 +/- 7.81) g/g prot (P < 0.05) in the control. Epimedium significantly increased sperm viability in the treatment group (60.03 +/- 7.54)% as compared with the model rats (69.90 +/- 12.58)% (P < 0.05).

CONCLUSIONEpimedium can lessen chemotherapy-induced damage to the epididymis and protect the reproductive function of rats.

Animals ; Antineoplastic Agents ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Epididymis ; drug effects ; physiopathology ; ultrastructure ; Epimedium ; chemistry ; Infertility, Male ; chemically induced ; physiopathology ; prevention & control ; Male ; Microscopy, Electron, Transmission ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

This study was aimed to prepare a reconstructed B. Fragilis-derived recombinant α-galactosidase developed for human B to O blood group conversion. Based on the construction of recombinant E. Coli (DE3) which can express α-galactosidase, the inducing time and inducer concentration were optimized for high expression of α-galactosidase. Then, the expression products in supernatant were purified by cation and anion exchange column chromatography. The purified α-galactosidase was used to treat B group red blood cells in phosphate buffer (pH 6.8) for 2 hours to prepare O group red blood cells. The results showed that the optimal inducing conditions for α-galactosidase expression were IPTG 0.1 mmol/L, 37°C and 2 hours. The specific enzyme activity of purified protein increased from 0.42 U/mg to 2.1 U/mg as compared with pre-purification. And, the conditions of B to O blood group conversion were 26°C, pH 6.8 (neutral pH condition) and 2 hours. Moreover, 225 µg of the enzyme could converse 1 ml B red blood cells to O completely. It is concluded that the technology of expression and purification of recombinant α-galactosidase has been established, and the purified protein can converse B red blood cells to O completely, which means that an effective enzyme conversing B red blood cells to O has been obtained.
ABO Blood-Group System ; immunology ; Bacteroides fragilis ; enzymology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Humans ; Recombinant Proteins ; biosynthesis ; alpha-Galactosidase ; biosynthesis

BACKGROUND: There are various treatment methods for osteonecrosis of the femoral head (ONFH) and collapse, but conservative treatment is invalid. Once femoral head collapse occurs, the development is irreversible. Our previous research has shown that local administration of zoledronic acid can prevent necrotic femoral head collapse. Moreover, bone marrow mononuclear cells obtain satisfactory short-term efficacy in the treatment of ONFH. OBJECTIVE: To investigate the curative efficacy of local administration of mononuclear cell, platelet-rich plasma and zoledronic acid for the prevention and treatment of early ONFH and collapse. METHODS: This prospective, single-center, randomized, parallel, controlled clinical trial was conducted at the Chinese PLA General Hospital, Beijing, China. One hundred patients with ONFH (stages I-II by Ficat and Arlet classification) were enrolled and randomly assigned into either the treatment group or control group (n=50 per group). Patients were given an injection of mononuclear cell, platelet-rich plasma and zoledronic acid into the necrotic femoral head, or drilling decompression at the necrotic area. Patients in both groups were then followed up for 4, 8, 12, and 18 months. The primary outcome measures were the blood supply, osteogenesis and appearance of the necrotic femoral head observed on hip perfusion by dynamic MRI, CT restruction of the hip joint and radiography of the hip joint, as well as Harris hip scores and numerical rating scale scores. Secondary outcome measures included SF-36 Health Survey and Activities of Daily Living scores. DISCUSSION: The outcomes of this trial have provided quantitative data for analyzing the effectiveness of local administration of mononuclear cell, platelet-rich plasma and zoledronic acid on ONFH and collapse. Written approval for this protocol was obtained from the Ethics Committee of the Chinese PLA General Hospital in China (approval No. S2015-082-01). Participants and their families are informed of the study protocol and procedures, and signed an informed consent. The study was in accordance with the guidelines of the Declaration of Helsinki, formulated by the World Medical Association. Trial began in January 2015 and will be completed in December 2017. Trial results will be published in scientific reports, or in peer-reviewed journals. This trial was registered with the ClinicalTrials.gov identifier: NCT02721940. Patient recruitment is ongoing.

OBJECTIVETo study the expression of recombinant human SCF-TPO fusion protein and its biological function.

METHODSFour primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.

RESULTSThe high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.

CONCLUSIONSExpressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; physiology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics ; metabolism ; physiology ; Thrombopoietin ; genetics ; metabolism ; physiology