OBJECTIVE: To extract effective constituent of Magnolia offcinalis and to identify its structure.METHODS: Ultrasound extraction was used to extract magnolol and honokiol from M.officinalis and the extracts were isolated using polyamide column chromatography.The content of magnolol and honokiol were determined using RP-HPLC.RESULTS: The optimal extractive condition was moderate ultrasonic,95% alcohol,8-fold solvent,extracting for three times,each time for 30 min.The extraction rates of phenols,magnolol and honokiol were 1.92%,1.26% and 0.66%,respectively.Honokiol was obtained by polyamide column chromatography.CONCLUSION: The method is simple,stable and supply reference for the extraction and identification of M.officinalis.

Objective To investigate the possibility of early diagnosis of colorectal cancer by combined detec-tion of tumor markers dermcidin(DCD),CA199,CA724 and CEA. Methods We selected 81 colorectal cancer patients and 30 healthy individuals. Serum levels of DCD were detected by ELISA,and serum levels of CA199, CA724 and CEA were detected by electrochemical luminescent immunoassay,and the expression of DCD in colorectal cancer tissue and para-carcinoma tissue was measured by Western blot. Results Serum levels of CA199,CA724,and CEA were significantly higher in colorectal cancer patients than in healthy individuals(P<0.001). Combined detection of DCD,CA199,CA724 and CEA had a greater AUC(0.942)for identifying colorec-tal cancer from the controls. The expression of DCD was significantly higher in colorectal tissue than in para-carci-noma tissue(P<0.05). Conclusions Level of DCD expression was significantly higher in colorectal tissue than in para-carcinoma tissue. Expressions of serum DCD,CA199,CA724,and CEA may be a marker in diagnosis of can-cer,with higher clinical value for the adjuvant diagnosis of colorectal cancer.

Objective To analyze the outcome of intra-artery chemotherapy for T1G3 bladder cancer , and its effectiveness and safety. Methods From June 2003 to May 2014, 39 patients with T1G3 bladder cancer chose intra-artery chemotherapy (Gemcitabine plus cis-platin), and close follow-up was required after 2 cycles of chemotherapy. During the follow-up, transurethral resection of bladder tumor was performed for non-muscle invasive bladder cancer, and cystectomy was performed for muscle invasive tumor. Results Of all patients, 32 were male and 7 were female. The median age was 56 years old (range: 32-82 years), and median follow-up time was 56 months (range: 12-136 months). Nineteen patients were primary bladder cancer, and 20 were recurrent tumor. During the follow-up, 17 patients developed recurrent tumors, including 8 progressed tumors and 3 died from tumor. Two-year and 5-year progressed-free survival were 88% and 74%, and 2-year and 5-year cancer-specific survival were 97% and 89%, respectively. During 5 years′ follow-up, 81% survivor preserved intact bladder, and only 1 patient cancelled chemotherapy for adverse effect. Conclusions Intra-artery chemotherapy (GC regimen) is a choice for T1G3bladder cancer, preventing from disease progression with good tolerance.

BACKGROUND: Chronic osteomyelitis is difficult to manage clinically, and two or more operations were commonly needed. No satisfactory method for one-stage repair has been currently available.OBJECTIVE: To examine the possibility of using fibrin glue(FG) as the common carrier for both bone morphogenetic protein(BMP) and gentamicin for one-stage repair of infected bone defects.DESIGN: A completely randomized controlled experiment.SETTING: Center of Orthopaedic Surgery, Lanzhou General Hospital of Lanzhou Area Command of of Chinese PLA.MATERIALS: The experiment was conducted using 48 healthy adult Chinchilla rabbits of either sex on normal diet with body mass of 1.9 to 2.4 kg,provided by the Institute of Biological Products, Lanzhou, Gansu Province.INTERVENTIONS: Animal models of chronic osteomyelitis were established in 48 Chinchilla rabbits, and after thorough debriding of the wounds,an elongated semicircular bone defect was induced 1.5 cm to the medial proximal metaphysis in the tibias. Three treatments were subsequently given with FG, MP and gentamicin complex(Group A), FG/BMP complex(Group B), and nothing(control group, Group C), respectively.MAIN OUTCOME MEASURES: The general conditions of the rabbits were observed postoperatively. Bacterial culture of the bone specimen, bacterial counting, X-ray and histological examination were performed.RESULTS: Rabbits in Group A exhibited obviously better infection control and greater regenerated bone mass than those in Group B, and no significant difference was noted in infection control between Groups B and C, the latter group showing poor bone defect repair.CONCLUSIONS: FG, BMP and gentamicin complex helps to control infection and promotes bone formation, and can be used to repair bone defects caused by infection and open injury with severe contamination.

Objective To detect the effect of low-dose heparin on endothelial function by detecting the change of yon Willebrand factor (vWF) in blood plasma of patients with sepsis shock after treatment with low-dose heparin in order to observe organ function and prognosis of patients.Methods A total of 118 patients with sepsis shock were randomized into low-dose heparin group and control group.In addition to the routine treatment,the patients in low-dose heparin group were given low-dose heparin,while those in control group were not.The levels of APTT,PT,platelet (PLT) count,hepatic function and renal function were determined before and after treatment in two groups,and hemorrhagic events or other complications were recorded.The lengths of stay in ICU and in hospital,the days of mechanical ventilation and the rates of acute respiratory distress syndrome (ARDS),disseminated intravascular coagulation (DIC) and multiple organ dysfunction syndrome (MODS) and 28-day survival rate in the two groups were documented.The levels of vWF in patients of two groups were determined by using immunoturbidimetry before and after treatment.Results The rates of DIC and MODS in low-dose heparin group decreased significantly after therapy (rate of DIC,10.6％ vs.26.7％,P ＜ 0.05;rate of MODS,19.3％ vs.32.0％,P ＜ 0.05).Hepatic function and renal function in low-dose heparin group were significantly improved than that in the control group (P ＜ 0.05),28-day survival rate in the low-dose heparin group was higher than that in the control group (78.3％ vs.69.1％,P ＜0.05),the differences between low-dose heparin group and control group were not statistically significant in the lengths of stay in ICU and hospital,the days of mechanical ventilation and the rate of ARDS (P ＞ 0.05).The differences in APTT,PT and PLT were not significant from pre-treatment to after treatment and between the two groups (P ＞ 0.05).The levels of vWF in low-dose heparin group decreased significantly after therapy rather than those in control group (P ＜ 0.05).Conclusions The endothelial function is improved in patients with sepsis shock after employment of lowdose heparin.Low-dose heparin alleviates the interaction between coagulation and inflammation,improves hepatic,renal and other important organs function,decreases the rate of MODS and increases 28-day survival rate patients with sepsis.The low-dose heparin therapy is a safe and promising treatment in sepsis patients without severe side effects.

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

Objective To understand the prevalence of alcohol dependence in individuals living in Guangxi Zhuang Autonomous Region,China. Methods From July 2007 to December 2007,21290 subjects aged more than 15 were randomly selected for 6 Prefecture-level cities and 10 counties by stratified multi-stage cluster sampling method. All subjects were investigated by face to face interview. The Chinese version of Composite International Diagnostic Interview (CIDI3.0) Provided by the Chinese Center for Disease Control And Prevention was used as the screening tool and the diagnoses were made with the 10th edition of international classification of Disease Criteria. Results 18 219 subjects completed the screening,the Completion rate was 85.58‰. The general point prevalence and the general lifetime prevalence of alcohol dependence in individuals living in Guangxi was both 4.50‰. The general lifetime prevalence was higher in rural (6.17‰) than in urban (1.85‰) ( RR =0.31,95% CI =0. 17 ～0.57, P=0. 000) ,higher in men (8.59‰) than in women (0. 33‰) ( RR =0.04,95% CI =0. 01 ～ 0.13, P = 0.000 ) and higher in Zhuang (9.63‰) than in Han(1.95‰) ( RR = 5. 01,95% CI = 3.02 ～8.34, P = 0.000). The lifetime prevalence of alcohol dependence tends upwards as age increasing (P＜0.05 ) and it differs in different marital status,higher in divorce groups (P ＜ 0.05). Conclusion Schizophrenia, Depression disorder, Alcohol dependence disorder are the most common mental disorders and have become the public health problem in Guangxi. To do this,it is necessary to carry out targeted research in prevention and rehabilitation.

Objective:To evaluate the effectiveness and safety of 3.5% lidocaine hydrochloride ophthalmic gel for eye surface anesthesia.Methods:A multicenter, randomized, double-blind, placebo controlled clinical trial was conducted in 10 hospitals in China from August 2018 to April 2019 under the approval of an Ethics Committee of Beijing Hospital (No.2018BJYYEC-014-02). A total of 220 fellow eyes of 220 subjects who received ocular surgery in one eye were actually enrolled according to a same included criteria in different institutes.The 220 eyes were randomly divided into control group and trail group.Two drops of lidocaine hydrochloride gel were dropped at about 5 mm away from corneal limbus at 6 o'clock direction of experimental eye of the trail group, and the blank gel was used in the eyes of the control group in the same way.The pain sensation was assessed with a 0.3 mm toothless micro forceps on conjunctiva within a specified time, and ''pain'' or ''no pain'' was answered by the subjects.The primary effective indexes, namely the number of eyes and percentage of ''no pain'' within 5 minutes following dropping, as well as the secondary indexes including the onset time point of the drug and the duration of anesthesia were recorded and evaluated.Safety evaluation took ocular and system adverse events into account.Results:Within 5 minutes after dropping, ''no pain'' occurred in 104 eyes (94.55%) and 29 eyes (26.36%) in the trail group and control group, respectively, showing a significant difference between the two groups ( χ2=106.948, P<0.001). And there was a significant difference in anesthesia onset time between the trail group and control group (40.0 seconds vs. 300.0 seconds) ( Z=-15.17, P<0.001). The duration of anesthesia was 860.5 (577.5, 1 180.0) seconds in the trail group and 676.0 (280.0, 1 401.0) seconds in the control group, with no statistically significant difference between the two groups ( Z=0.898, P>0.05). The incidence of adverse events in the trail group and control group were 5.45% (6/110) and 4.55% (5/110), respectively, without statistical significance between them ( P=1.000). Conclusions:The 3.5% lidocaine hydrochloride is a safe, effective, easy to use and high-quality surface anesthesia drug for eye surgery.

Next-generation sequencing has yielded a vast amount of cattle genomic data for global characterization of population genetic diversity and identification of genomic regions under natural and artificial selection. However, efficient storage, querying, and visualization of such large datasets remain challenging. Here, we developed a comprehensive database, the Bovine Genome Variation Database (BGVD). It provides six main functionalities:gene search, variation search, genomic sig-nature search, Genome Browser, alignment search tools, and the genome coordinate conversion tool. BGVD contains information on genomic variations comprising ～60.44 M SNPs, ～6.86 M indels, 76,634 CNV regions, and signatures of selective sweeps in 432 samples from modern cattle worldwide. Users can quickly retrieve distribution patterns of these variations for 54 cattle breeds through an interactive source of breed origin map, using a given gene symbol or genomic region for any of the three versions of the bovine reference genomes (ARS-UCD1.2, UMD3.1.1, and Btau 5.0.1). Signals of selection sweep are displayed as Manhattan plots and Genome Browser tracks. To further investigate and visualize the relationships between variants and signatures of selection, the Genome Browser integrates all variations, selection data, and resources, from NCBI, the UCSC Genome Browser, and Animal QTLdb. Collectively, all these features make the BGVD a useful archive for in-depth data mining and analyses of cattle biology and cattle breeding on a global scale. BGVD is publicly available at http://animal.nwsuaf.edu.cn/BosVar.

Objective:To investigate the therapeutic efficacy and potential mechanisms of integrin α vβ 3-targeted radionuclide therapy (TRT) in combination with anti-programmed cell death protein ligand 1 (PD-L1) immunotherapy. Methods:Integrin α vβ 3-targeted molecule Arg-Gly-Asp (RGD) was conjugated with Evans blue (EB) and then labeled with 177Lu to obtain 177Lu-EB-RGD. The radioactivity and radiochemical purity were determined. MicroSPECT imaging, biodistribution, and in vivo therapeutic efficacy were subsequently performed in MC38 murine colon cancer models. Volume of tumor and body mass of mice were observed to assess the therapeutic efficacy and safety ( n=9 in each group). Flow cytometry was used to evaluate therapy response of saline-treated (control, group A), 18.5 MBq 177Lu-EB-RGD-treated (group B), 10 mg/kg PD-L1 antibody-treated (group C), TRT combined with immunotherapy-treated (group D, 18.5 MBq 177Lu-EB-RGD and 10 mg/kg PD-L1 antibody) mice and alterations in tumor microenvironment (PD-L1 + immune cells, CD8 + T cells and regulatory T cells). Independent-sample t test and repeated measures analysis of variance were used for data analysis. Results:The radioactivity of 177Lu-EB-RGD was (55.85±14.00) GBq/μmol. SPECT imaging clearly visualized the MC38 tumors in mice models with high uptake and long retention time, the tumor/muscle ratio reached 14.87±0.88 at 24 h postinjection, while less uptake and retention in normal tissues. Tumor uptake of 177Lu-EB-RGD was significantly higher than that of 177Lu-RGD 4 h post-injection ((12.00±1.60) vs (3.69±0.37) %ID/g; t=8.63, P<0.01). The efficacy results between each treatment group was significantly different ( F=7.32, P=0.03) at day 6 post-treatment. The combination therapy showed the most outstanding anti-tumor efficacy with 7/9 mice showed complete response. Flow cytometry results showed that TRT up-regulated the PD-L1 expression significantly, namely, PD-L1 + immune cells in group B and group A were significantly different (CD45 + /PD-L1: 2.34% vs 0.95%, CD11b + /PD-L1: 2.41% vs 0.66%; t values: 11.17 and 8.70, both P<0.01); immunotherapy and combination therapy dramatically stimulated the infiltration of CD8 + T cells (2.07% vs 0.26%, 2.71% vs 0.26%; t values: 4.10 and 6.03, both P<0.05). Conclusion:TRT in combination with immunotherapy synergistically enhance anti-tumor efficacy, which is expected to play a role in the treatment of patients with advanced tumor where TRT can be applied.