OBJECTIVE:To establish the method for determining 10 residual organic solvents in norvancomycin hydrochloride raw material. METHODS:Headspace gas chromatography was performed on the column of nitro modified polyethylene terephthal-ate glycol as stationary phase capillary column;the oven temperature program started at 40 ℃ for 3 min and increased at a rate of 8 ℃/min up to 150 ℃ for 10 min;the temperature was 200 ℃ with carrier gas of high-purity nitrogen gas,the constant flow rate was 5 ml/min with split ratio of 15∶1;the headspace vial equilibrium temperature was 85 ℃ with equilibrium time of 40 min,and the volume was 1 ml. RESULTS:The concentration of n-pentane,acetone,ethanol,benzene,acrylonitrile,toluene,xylene,chlo-robenzene,styrene,divinylbenzene had good linear relationship with its peak area values(r=0.995 7-0.999 9);the RSDs of preci-sion,repeatability tests was ≤6.6%;average recovery was in the range of 94.3%-106.6%(RSD=0.5%-4.5%,n=9). CONCLU-SIONS:The method is fast,sensitive and accurate,and can be used for the determination of residual organic solvents in norvanco-mycin hydrochloride raw material.

Objective To evaluate the efficiency of transcatheter closure of secundum artrial septal defect (ASD) using Amplatzer occluder device in 5 years.Methods There were 18 children with ASD.Each ASD was occluded using Amplatzer occluder device through the percutaneous procedure.The closure procedure was guided by fluoroscopy and transthoracic echocardiography.All patients were followed up by physical examination,color flow echocardiography,chest radition,electrocardiography at 48 h,1month,3 month,6 month,one year,two years,three years,four years and five years.Results Cardiac murmur disappeared or lightened.Arrhythmia and right heart volume in all patients were improved after the procedure.A case revealed a trival shunt of short duration and vanished after one month color flow echo.A case has “spring′s beat” in heart when she goes to bed in three years after the procedure.Another patient had serious headach after closure and recoved after heparin therapy.Conclusions Transcatheter closure of secundum ASD using amplatzer occluder device is safe and efficient by follow-up.In order to prevent cerebral embolism,heparin should be injected in vein in two days.

To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

OBJECTIVE: To investigate the anti-arthritic effect of bee venom in adjuvant induced arthritis (AIA) in rats. METHODS: Thirty-two male SD rats were enrolled in the experiment. Six were treated as negative controls and 20 AIA models were randomly divided into 3 groups: model controls (n=6), sodium chloride treatment group (n=6), and bee venom treatment group (n=8). The rats in the model control were killed before the treatment and the peripheral blood and synovium samples were collected for pre-treatment controls. The rats in the bee venom treatment group were injected hypodermically with bee venom for 14 days, while those in the sodium chloride treatment group were treated with the same volume of sodium chloride. During this period, the circumference of the affected joints and the total scores of the joints in all groups were measured every 2 days and X ray examinations were performed before and after the treatment. At the end of the treatment, all the rats were killed and their peripheral blood and synovium samples were collected for measurements of tumor necrosis factor TNF-alpha and interleukin IL-1 beta and histological studies, respectively. RESULTS: Compared with the sodium chloride group, the rats in the bee venom treatment group were less swollen in joints and circumference of joints and lower joint scores decreased (P<0.05). At the same time, the bone erosion and the infiltration of inflammatory cells in the synovium were also significantly reduced in the bee venom treatment group. In addition, the serum concentrations of TNF-alpha and IL-1 beta were significantly lower in rats of the bee venom treatment group than those of the sodium chloride group (P<0.05 and P<0.01, respectively). CONCLUSION Bee venom is effective in treating AIA by reducing synovitis, downregulating the serum concentrations of cytokine TNF-alpha and IL-1 beta and alleviating the bone erosion.
Animals ; Arthritis, Experimental ; drug therapy ; Bee Venoms ; therapeutic use ; Interleukin-1beta ; blood ; Male ; Rats ; Rats, Sprague-Dawley ; Synovial Membrane ; pathology ; Synovitis ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

Objective: To explore the time and genotype distribution of human enterovirus (HEV) isolated from sewage in Shanghai in 2013-2014. Methods: One sewage sample each was collected from two local sewage plants located in Minhang District and Jiading District on the same day at the day 24-28 of every month from 2013 to 2014. Each sample weighed 1 L. The specimens were concentrated by anionic membrane absorption, eluted with beef extract solution, and then used to inoculate RD, HEp-2, and L20B cell lines. A total of 249 enterovirus strains were isolated from sewage samples during the study period, including 185 non-polio enterovirus (NPEV) and 64 poliovirus (PV) strains, which were identified as vaccine strains. RT-PCR and Sanger sequencing were performed to identify HEV genotypes. Homologous analysis of VP1 sequences was conducted using BioEdit (version 7.0.0). Phylogenetic analysis was performed using the neighbor-joining method based on the alignment of VP1 gene sequences using MEGA (version 4.0.2). Results: Among 185 NPEV strains, 178 strains were successfully sequenced and classified into 15 genotypes, including coxsackievirus group B (CVB) 2, 3, and 5; enteric cytopathic human orphan (ECHO) virus 1, 3, 6, 7, 11, 13, 19, 20, 24, 25, and 30; and coxsackievirus group A 4. CVB5 and ECHO6 genotypes accounted for 33.5% (56 strains) and 24.9% (43 strains) of NPEV isolates, respectively. During the study period, HEV isolates were mainly isolated in summer and autumn in Minhang District. ECHO6 strains were frequently isolated from June 2013 to July 2014. Thereafter, the number of ECHO6 strains gradually reduced in the second half of 2014. CVB5 strains demonstrated scattered distribution from 2013 to the first half of 2014 and gradually increased in the second half of 2014. The distribution of ECHO6 and CVB5 strains in Jiading District was similar to that in Minhang District. In 2013-2014, CVB5 strains comprised C6 and C8 subgenotypes, which belong to two transmission chains and show large differences compared with foreign strains isolated during the same period. ECHO6 strains comprised C6, C8, and D9 subgenotypes, which belong to three transmission chains. Moreover, ECHO6 subgenotype D9 was a dominant subgenotype in Shanghai, with broad geographical distribution both at home and abroad. Conclusion Poliovirus was identified as a vaccine strain in environmental surveillance from June 2013 to April 2014 in Shanghai. Several transmission strains of ECHO6 and CVB5 were identified, which were the dominant serotypes.

OBJECTIVETo compare the clinical and oncological outcomes between laparoscopic and open intersphincteric resection in patients with low rectal cancer.

METHODSFrom January 2007 to January 2010, patients with low rectal cancer treated by laparoscopic or open intersphincteric resection were included in a retrospective comparative study. Patients were classified into laparoscopy group (n=27) and open group (n=41). The operative procedures, postoperative complications, anal function and clinicopathological data were compared.

RESULTSCompared to the open group, the laparoscopic group had longer operative time [(242.2±42.5) min vs. (199.1±44.3) min, P=0.000], less blood loss [(150.5±102.2) ml vs. (258.4±149.2) ml, P=0.002], faster recovery of bowel function [(2.9±1.1) d vs. (3.6±1.5) d, P=0.032] and resumption of regular diet [(6.6±1.2) d vs. [(7.5±1.7) d, P=0.012], and shorter postoperative hospital stay [(7.7±1.4) d vs. (9.1±2.4) d, P=0.006]. The postoperative complication rate between the laparoscopic and open groups was not significantly different [18.5% (5/27) vs. 19.5% (8/41), P=0.464]. Oncological parameters were comparable between the two groups including lymph node harvested [(14.1±4.1) vs. (16.4±6.8), P=0.113], distal resection margin [(1.4±0.7) cm vs. (1.6±0.8) cm, P=0.311], and circumferential margin [7.4% (2/27) vs. 2.4% (1/41), P=0.709]. Local recurrence rates in laparoscopic and open groups were 7.4% (2/27) and 2.4% (1/41), and distant metastasis rates were 0 and 4.9% (2/41) respectively, and the differences were not significant (both P>0.05).

CONCLUSIONSLaparoscopic intersphincteric resection possesses same efficacy of open intersphincteric resection with less blood loss, shorter recovery time and hospital stay, and similar oncological outcomes, and no increased postoperative morbidity and mortality.

Aged ; Female ; Humans ; Laparoscopy ; Laparotomy ; Male ; Middle Aged ; Prognosis ; Rectal Neoplasms ; surgery ; Retrospective Studies

OBJECTIVETo prepare the trimeric subunits of recombinant human mannan-binding lectin (MBL) with biological activities.

METHODSA prokaryotic expression vector containing human MBL N-terminal deletant (rhMBLδN) gene we previously constructed was transformed into E. coli for efficient expression of rhMBLδN fusion protein. Based on the principle that the collagen polypeptides tend to self-assembly into the tertiary structure of proteins by forming a triple helix due to the characteristic properties of the collagen proteins, rhMBLδN fusion protein was limitedly hydrolyzed with thrombin. The obtained rhMBLδN polypeptide was repeatedly dialyzed in 50 mmol/L PBS (pH7.2) and ddH(2)O, and the final product was analyzed for its bioactivities using a ligand-binding assay and a C4d deposition assay.

RESULTSrhMBLδN polypeptide with a relative molecular mass of about 20 000 was obtained by limited proteolysis of rhMBLδN fusion protein with thrombin. Repeated dialyses of rhMBLδN polypeptides in 50 mmol/L PBS and ddH(2)O resulted in the isolation of the trimeric subunit trhMBLδN (with a relative molecular mass of about 50 000), which contained a collagen-like helix. The trhMBLδN protein had a higher ligand-binding activity than rhMBLδN polypeptide, and acquired the activity to initiate the lectin pathway of complement activation, but the activities were lower than those of natural MBL.

CONCLUSIONWe have successfully obtained the bioactive trimeric subunit of rhMBL, trhMBLδN, and this structural subunit is also the functional subunit of the MBL molecule.

Complement Activation ; Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Mannose-Binding Lectin ; biosynthesis ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis

OBJECTIVETo study the chemical constituents in the root of Isatis indigotica.

METHODThe constituents root were separated through various chromatographic techniques and their structures were elucidated by means of physicochemical properties and the analysis of their spectral data.

RESULTEleven compounds were isolated and identified as (+) -isolariciresinol (1), lariciresinol (2), lariciresinol-9-O-beta-D-glucopyranoside (3), lariciresinol-4'-O-beta-D-glucopyranoside (4), lariciresinol-4,4'-bis-O-beta-D-glucopyranoside (5), 3-formylindole (6), 1-methoxy-3-indolecarbaldehyde (7), 1-methoxy-3-indoleacetonitrile (8), deoxyvasicinone (9), epigoitrin (10), adenosine (11).

CONCLUSIONCompounds 4-8 were isolated from I. indigotica for the first time.

Furans ; analysis ; chemistry ; isolation & purification ; Glucosides ; analysis ; chemistry ; isolation & purification ; Indoles ; analysis ; chemistry ; isolation & purification ; Isatis ; chemistry ; Lignans ; analysis ; chemistry ; isolation & purification ; Lignin ; analysis ; chemistry ; isolation & purification ; Naphthols ; analysis ; chemistry ; isolation & purification ; Plant Extracts ; analysis ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To investigate the epidemiological status of visceral leishmaniasis in Hamangou coal mine area of Korla City of Xinjiang Uygur Autonomous Region.Methods Based on a hint of possible existence of patients, a retrospective survey was carried out house by house to find cases with suspected signs/symptoms of the disease.Meanwhile, a survey on current status was conducted, including physical examination(liver and spleen palpation) to those less than 15 years-old, leishmanin skin test and rk39 immunochromatographic strip test for part of the residents.Bone marrow smears were examined for the cases with clinical signs/symptoms and positive rk39 strip test.Sandflies were collected using routine methods in and around the area, identified, and dissected to find infection of promastigotes.Results Leishmanin skin test was performed in 185 people with a positive rate of 21.1%(39/185), 39 out of 140 local residents who have lived there for more than 6 years showed positive(27.9%) , while all residents who have lived less than 6 years and children under 5 years old were negative.Of the 81 children under 15 years old with a negative skin test, one showed positive for rk39 strip test, and leishmania body was found in the bone marrow smear of this case, so confirmed as visceral leishmaniasis.12 sandfies were identified as Phlebotomus alexandri, and natural infection with promastigotes was found in one sandly.Conclusion The investigation confirms that visceral leishmaniasis is endemic in the Hamangou coal mine area.

OBJECTIVEVasoactive intestinal peptide (VIP) is a neuro-peptide that can modulate immunity. Previous studies indicated that VIP can attenuate the deleterious consequences of severe sepsis and septic shock by regulating production of inflammatory cytokines in immune activated cells. The signaling induced by bacterial components occurs primarily through Toll like receptors (TLRs). TLRs have been recognized to play a key role in pathogen recognition and innate immunity. It was convincingly demonstrated that lung is one of early suffered disaster organ and may trigger multiple organ dysfunction syndrome in sepsis. The present study was conducted to investigate the effects of VIP on TLR2/4 mRNA expressions on acute lung injury of endotoxic shock induced by lipopolysaccharide (LPS) in rat.

METHODForty Sprague-Dawley rats were randomly divided into 3 groups, i.e., LPS shock group (n = 16), LPS + VIP group (n = 16), and control group (n = 8). LPS shock model was established by LPS (E. coli O(55)B(5) 10 mg/kg) with tail intravenous injection. The rats in LPS + VIP group were given a bolus of 5 nmol VIP intravenous injection follow by LPS. The rats in control group were given normal saline. The rats were sacrificed at 6 h, 24 h after being injected. The lung tissues were collected. The TLR2 mRNA and TLR4 mRNA expressions were detected by RT-PCR from the lung tissues. Pathological changes of the lungs were observed by light microscope and electron microscope 24 h after LPS injection.

RESULT(1) Lung histopathology: the alveolar space was full with leukocyte, necrotic cells, segmental hemorrhage and protein effusion. Partial alveolar space was enlarged, lung interstitial edema were observed in LPS shock group. However, pathological changes of LPS + VIP group were milder than those in LPS shock group. (2) The expressions of TLR2 mRNA and TLR4 mRNA were significantly higher in LPS shock group compared with those of the control group (F = 16.638, P = 0.000; t = 5.876, P = 0.000), TLR2 mRNA and TLR4 mRNA expression on 24 h was down-regulated in LPS + VIP shock subgroup than those in LPS shock subgroup (F = 16.676, P = 0.000; t = 3.946, P < 0.001).

CONCLUSIONExpressions of TLR2 mRNA and TLR4 mRNA were up-regulated on LPS induced lung injury in rats. VIP mitigated lung injury induced by LPS, which may be related to TLR2 mRNA and TLR4 mRNA down-regulation of expression. The effect of VIP may suggest a protective mechanism in sepsis. VIP may play a potential protective role in severe infection.

Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Down-Regulation ; Lipopolysaccharides ; Lung ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Up-Regulation ; Vasoactive Intestinal Peptide ; pharmacology