Objective:To prepare standard decoction of Saposhnikoviae Radix pieces,and conduct a study on the quality standards, in order to provide the scientific reference for the study of clinical application and other decoction standard decoctions. Method:By the traditional water decocting method,11 batches of standard decoction of Saposhnikoviae Radix pieces were prepared to determine pH,prim-O-glucosylcimifugin and 4'-O-β-D-glucosyl-5-O-methylvisamminol content,calculate the paste rate and transfer rate,and investigate the feasibility of the measurement method. An analytical method for HPLC fingerprint was established. Hypersil ODS2 column (4.6 mm×250 mm,5 μm) was adopted and eluted with methanol-water as the mobile phase in a gradient mode. The detection wavelength was 254 nm,the flow rate was 1 mL·min^-1,and the column temperature was 25℃. The fingerprint analysis method was used to evaluate the similarity of the 11 batches of standard decoction of Saposhnikoviae Radix pieces. Result:Through the determination of 11 batches of standard decoctions of Saposhnikoviae Radix pieces,the average pH value was 5.5,the paste-out rates were between 34.3% and 46.3%,and the average paste-out rate was 41.4%,the standard deviation was 3.7%, the transfer rate of prim-O-glucosylcimifugin was 66.8%-93.5%,the average metastasis rate was 79.4%,the standard deviation was 12.1%, the transfer rate of 4'-O-β-D-glucosyl-5-O-methylvisamminol was 70.4%-98.2%,the average transfer rate was 83.40%,the standard deviation of 10.8%. The results of various methodological studies were consistent with the requirements of RSD. The fingerprinting analysis was performed using similarity evaluation software for traditional Chinese medicine fingerprints,and 9 principal common peaks were identified. The similarity was higher than 0.9. Conclusion:The quality evaluation method established in this study has a good stability,precision and repeatability. The fingerprints have a high similarity,which is suitable for the quality evaluation of standard decoctions of Saposhnikoviae Radix pieces, and can provide the scientific basis for the quality evaluation of other related preparations.

OBJECTIVETo study the status quo of oral professional and technical personnel of Guyuan city, Ningxia.

METHODSIn consultation with Administrative Department of Public Health, all oral health institutions by age, gender, education, professional, and department in four counties (Longde, Jingyuan, Pengyang and Xiji) and Yuanzhou district in Guyuan City were investigated. The distribution, basic information, professional branch, ratio with the amount of health system medical staff and population were analyzed.

RESULTSAll comprehensive hospital at the county level and above set up department of stomatology. Center for Disease Control and Prevention, health inspection bureau, maternity and child health care bureau and 78.46% town health center didn't set up department of stomatology. Oral professional and technical personnel of Guyuan city in number was 115 and mainly young doctors, men were more than women. On education, the tertiary education was mostly received. On professional title, number of middle and senior professional title was relatively small. On profession, multi-disciplinary was common. The ratio of oral professional to general medical staff were 1:30, to local population was 1:13 thousands.

CONCLUSIONThe education and professional title of oral professional and technical personnel should be improved. It is recommended that comprehensive hospitals above the county level should branch in Stomatology, Center for Disease Control and Prevention, health inspection bureau, maternity and child health care bureau and town health center should offer services of stomatology, at least provide one dental professional and technical personnel in each place.

China ; Dentistry ; manpower ; Female ; Humans ; Male

To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
Animals ; Antibodies ; genetics ; metabolism ; pharmacology ; Birds ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Hemagglutinins ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; drug effects ; immunology ; Influenza in Birds ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology

AIMTo investigate the protective effects and mechanism of IPC on myocardial ischemia/reperfusion injury.

METHODSEffects of IPC on arrhythmia and coronary blood flow and the release of AST, CPK, LDH, SOD and LFO at different time after ischemia/reperfusion injury in rat Langendorff hearts were studied.

RESULTSIPC decreased the release of AST, CPK and LDH and increased myocardial SOD activity and decreased LPO level. IPC also inhibited ischemia/reperfusion arrhythmias and increased coronary blood flow.

CONCLUSIONThe results showed that IPC had well protective effects on myocardial ischemia/reperfusion injury.

Animals ; Female ; Heart ; physiopathology ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Rats ; Rats, Wistar

Aptamers are a group of artificial oligonucleotides identified by exponential enrichment system evolution technology (Selective expansion of ligands by exponential enrichment, SELEX). Aptamers have been widely used in basic research, clinical diagnostics, and nano-technology. In this article we will introduce the technology of aptamer and summarize its applications in medical microbiology.
Aptamers, Nucleotide ; biosynthesis ; genetics ; Microbiological Techniques ; methods ; Microbiology ; SELEX Aptamer Technique ; methods ; trends

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

Bats are important reservoir animals and more than 60 viruses have been identified in bats with many of them highly pathogenic to human. In order to understand the natural background, genetic diversity of bat viruses in China and discover potential viral pathogens, Solexa sequencing based viral metagenomics focusing on bats tissues was established and to analyze the virome of bats collected from Jilin, Yunnan and Hunan province. By Solexa sequencing, 116 442 324 useful reads were obtained and assembled into 4 872 contigs, of which 8.2% (4 002/4 4872) were annotated to 36 viral families, including 19 vertebrate virus families, 6 plant virus families, 4 insect virus families and 4 phages. Further contigs analyses showed that some adenovirus, bocavirus, picobirnavirus, parvovirus contigs sequences were similar with known viruses. However, part of them shared limited identities to these viruses implying the discovery of new viruses. Moreover, PCR validation of adenovirus and bocavirus confirmed the results obtained by viral metagenomics. This study aimed to understand bat virome in China by viral metagenomics and could be helpful to establish effective surveillance on wildlife-associate zoonoses.
Adenoviridae ; genetics ; isolation & purification ; Animals ; Bunyaviridae ; genetics ; isolation & purification ; China ; Chiroptera ; virology ; Genome, Viral ; genetics ; Metagenome ; genetics ; Metagenomics ; methods ; Picornaviridae ; genetics ; isolation & purification

OBJECTIVETo discuss laparoscopic assisted radical D2 resection of distal gastric anatomy application ideas.

METHODSCollected the clinical data from January 2009 to January 2012 who underwent laparoscopic distal gastric resection in patients with D2 349 cases. There were 180 male and 169 female patients, and the age were (57 ± 3) years old (range 29-86 years), the body mass index of patients were (26.0 ± 2.0) kg/m(2) (range 20.5-32.8 kg/m(2)). The relevant surgical anatomy ideas had summarized.

RESULTSIn addition to 5 cases of obese patients with conversion to open, the remaining patients underwent laparoscopic distal gastric D2 resection. The operation is divided into 7 operating anatomical view. The operation time were 120-210 minutes and the blood loss were 50-200 ml. Postoperative complications occurred in 11 cases, including 5 cases of duodenal stump leakage, 2 cases of gastroparesis, 3 cases of small bowel obstruction, and abdominal bleeding in 1 case. All patients were discharged.

CONCLUSIONSUse zoning, exterior to interior of the anatomy, more conducive to master the operation of laparoscopic radical gastrectomy and standardized cleaning, to improve the operation efficiency and shorten the surgical learning curve and improve the quality of surgery has an important role.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastrectomy ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Stomach ; anatomy & histology ; Stomach Neoplasms ; surgery

OBJECTIVETo establish a method for isolation, cryopreservation and recovery of the highly viable human peripheral blood monomuclear cells (PBMNCs) so as to achieve the long-term preservation of PBMNCs.

METHODSA total of 80-100 ml peripheral blood were collected from the healthy volumteers aged over 50 years old. The PBMNCs were isolated by the Ficoll density gradient technique and cryopreserved gradually by program control method in liquid nitrogen freezer of -196 °C. The serum-free medium and autoloqous plasma medium were test for preservation of PBMNCs. The cell viability was assessed at time point of 1, 2, 4, 8, 12 and 24 months after thawing. Finally, the proliferation ability, purity and cytotoxicity were compared between the autologous immune lymphocytes (AIL) induced from cryopreserved PBMNCs and AIL as control from fresh PBMNCs.

RESULTSAfter separating, the cell viability was 99.6%±0.4%, and the recovery rate of lymphocytes was 58.4%±6.52%. The cell recovery rate of lymphocyte was 89.7%±3.82% at 24 months. The quality assurance program was reliable within 2 years of running. The AIL cells induced with cryopreserved PBMNCs were not significantly different from those induced from fresh PBMNCs in terms of proliferative action, purity and cytotoxicity(CD3(+)CD8(+) ≥45%,CD3(+)CD56(+) NKT≥10%,CD4(+)CD25(+) NKT≤10%).

CONCLUSIONManual separation of lymphocytes in vitro can get enough high-quality PBMNCs. The long-term cryopreserved PBMNC still maintain their high viability. The reinfusion of the clinical autologous immune cells would be advantageous for early tumor immunotherapy. Human AIL induced from cryopreserved PBMNC maintain their anti-tumor ability. These findings have the important implications for the application of these cells to adoptive cellular therapy.

Cell Survival ; Cryopreservation ; Humans ; Immunocompetence ; Leukocytes, Mononuclear

OBJECTIVETo investigate the efficacy of laparoscope complete mesocolic excision (CME) for right colon cancer.

METHODSThe clinical data of 49 cases of right colon cancer without metastasis and intestinal obstruction who underwent elective laparoscope CME by the same group of surgeons between October 2009 and June 2011 at the First Hospital of Jilin University were analyzed retrospectively.

RESULTSAmong the 49 cases with CME, the median number of total lymph node retrieved was 22. The positive rate of lymph node in patients with stage III disease was 16.3%. The median operative time of all the cases was 145 min. The mean intraoperative blood loss was 75 ml. The median time to flatus passage and defecation was 3 days and 6 days respectively. The median hospital stay was 12 days. The overall postoperative complication rate was 12.2% (6/49).

CONCLUSIONLaparoscope complete mesocolic excision for right colon cancer is safe and feasible.

Adult ; Aged ; Colectomy ; methods ; Colonic Neoplasms ; surgery ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies