OBJECTIVE:To guide rational use of Heamocoagulase agkistrodon for injection,and to provide reference for phar-maceutical care. METHODS:Treatment methods and outcome of 9 cases of allergic shock induced by Heamocoagulase agkistrodon for injection were retrieved from CNKI and VIP database during 2009-2015,and then analyzed statistically,based on one case of allergic shock induced by Heamocoagulase agkistrodon for injection in the Affiliated Hospital of Zhunyi Medical College. RE-SULTS:10 pationts with allergic shock induced by Heamocoagulase agkistrodon for injection mainly used the drug for hemostasis before and after surgery,heavy menstrual bleeding,etc.,with dose of 1 or 2 U. Route of administration included intravenous injec-tion and intravenous dripping. Allergic shock often occurred within 30 min after medication. CONCLUSIONS:The use of Heamoco-agulase agkistrodon for injection in some patients belongs to off-label drug use,and route of administration is different from that stated in package inserts. Allergic shock induced by Heamocoagulase agkistrodon for injection occurs rapidly;the drug should be used strictly according to package inserts and authoritative literatures;emergency plan should be developed before medication in or-der to guarantee the safety of drug use.

Objective To investigate the effectiveness of hyperthermia of different degrees in killing tumor cells and its influence on Na+ -K+ -ATPase activities in erythrocytes. Methods Cultured low differentiated stomach gland tumor cells (SGC-170) and large intestine gland tumor cells (LOVO) (1?106?ml-1 each) were mixed with concentrated RBCs respectively and incubated in warm bath of 37℃ , 42℃, 43℃ , 45℃ or 47℃ for 40 min respectively. After hyperthermic treatment tumor cells were isolated from RBCs using density gradient centrifugation and the live tumor cells were counted by typan staining. The isolated tumor cells were then cultured and the clone formation of tumor cells was checked. The cultured tumor cells were marked with 5-bromo deoxyuridine and DNA metabolism was examined. The Na+ -K+ -ATPase activities in RBC after hyperthermic treatment were also determined. Results The amount of tumor cells was significantly decreased by 40 min hyperthermic treatment in a temperature-dependent manner from 42-471 as compared with the control group (37℃) (P

Objective By comparing the proliferation and the human telomerase reverse transcriptase expression of bone marrow mesenchymal stem cells (BMSCs) from rheumatoid arthritis (RA) patients and normal human to explore the existence of BMSCs defects or variation in RA patients,and whether these cells could be used in autologous transplantation.Methods BMSCs were obtained from bone marrow of eight RA patients and healthy volunteers.The cells were isolated andcultured by gradient centrifugation and adherence cultivation,and the surface markers of BMSCs were identitfied by flow cytometertry.Firstly,BMSCs reproductive activity of the two groups were compared:the auxodrome and cell cycle of BMSCs were measured respectively,then the generation doubling time of cell colony and proliferation index were calculated in order to see whether there was a difference of the proliferation capability between the two groups.Furthermore,the hTERT were measured.Results BMSCs were no significant differences between morphology of RA patients and healthy control group.The cells were obtained proof MSCs by flow cytometry.The difference was not statistically significant (P＞0.05),RA group and the healthy control group P3-generation cell population doubling time (4.7±0.3) d,(4.8±0.3) d.The difference was statistically significant (P＜0.01),P6 generation of healthy control group cell population has doubling time (5.7±0.4) d,RA group (6.3±0.3) d,buut the group RA BMSCs population doubling time is longer than the healthy control group.The difference was not statistically significant (P＞0.05),on one hand of the healthy control group P3 BMSCs proliferation index (17.9±2.1)％,on the other hand RA group is (16.9±2.1)％.The difference was statistically significant (P＜0.01).P6-generation BMSCs proliferation index in healthy control group (10.7±1.8)％,RA group (6.2±2.2)％,RA group proliferation index than the control group.The difference was also significant (P＜0.01).The healthy control group P6 generation of MSCs hTERT expression was (10.6±1.5),RA group is (3.1±1.0),hTERT RA patients expression is lower compared with normal control group.Conclusion There is no significant difference in the proliferative capacity of the two groups at P3,but the BMSCs proliferation at P6 lags behind in the normal group in RA patients,which inferred that the senescences of the BMSCs of RA patients are earlier than healthy donors.The determination results of hTERT have shown that the difference between the two groups is statistically significant,and it proves that the premature aging may be associated with hTERT expression.

Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.

OBJECTIVE:To provide reference for clinical rational use of curcumae rhizoma. METHODS:According to litera-ture retrieval,the chemical structures,ingredient and efficacy difference of curcumae rhizoma with different sources and difference processed products were summed up. According to literature retrieval,telephone consultation and network retrieval,the production status of products with different sources and difference processing and the application in clinic and preparations in Jiangsu province were analyzed. RESULTS:The volatile oil content was the highest in Curcuma wenyujin;curcumin content was the highest in Cur-cuma phaeocaulis;the contents of 4 main active ingredients(curdione,curcumol,germacrone and curcumin)were relatively lower in Curcuma kwangsiensis,and there were quite different ingredients in the C. kwangsiensis from different producing areas. Results of investigating the annual output of medicinal materials in Sichuan,Guangxi and Zhejiang in 2016 showed,the annual output of C. kwangsiensis was the highest,and C. wenyujin and C. phaeocaulis were relatively lower. In the investigated 14 manufacturers producing curcumae rhizoma oil in China,12 took C. wenyujin and 2 took C. phaeocaulis as raw material. While the 9 manufactur-ers in Jiangxi province mostly took C. wenyujin(locally produced)as raw material,which was not from the genuine producing ar-ea-Zhejiang province. In the 10 third-grade class-A TCM hospitals and 20 retail TCM pharmacies in Jiangsu province,the C. phaeo-caulis and C. kwangsiensis were mainly used,in which,the use proportion of C. kwangsiensis accounted for about 70%,and C. wenyujin used little in clinic. The main processed product in clinic was curcumae rhizoma processed by vinegar,and crude product was rarely used. CONCLUSIONS:Currently,the clinical use of curcumae rhizoma shows problems in narrow selection of pro-cessed products and uneven quality,which can be solved by cultivating fine varieties in producing areas,increasing the planting amount of C. phaeocaulis,drug regulatory authorities'strict rules for processed products of curcumae rhizoma in hospitals and re-tail pharmacies.

Objective To evaluate the therapeutic effect of microwave ablation in combination with TACE for the treatment of primary liver carcinoma (PLC). Methods From Jan. 2004 to Dec. 2008, 63 PLC patients underwent ultrasound-guided microwave ablation (percutaneous or open) under general anesthesia. Repeated microwave ablation or TACE was used when an incompleted ablation or recurrence was found during postoperative regular follow-up. Results These 63 PLC patients have received a total of 82 sessions of microwave ablation procedure (1 to 5 sessions for each patient). There were 2 early postoperative deaths with a procedure-related mortality of 3.2%. At the end of the follow-up, 22 patients were alive and 38 died,and the other one was lost to follow-up. The survival rates in 1,2 and 3 years were 63.3%,42.1% and 26.5%, respectively, with a median survival of 20 months for all patients. The survival for PLC patients with early stage (TNM Ⅰ and Ⅱ) was significantly longer than that of advanced stage (TNM Ⅲ and Ⅳ). The 1,2 and 3 year's cumulative survival rate was 93.3%,86.7% and 65.0% respectively in those 15 cases with only single tumor and the diameter≤3 cm, which were significantly longer than that of other PLC patients. Of 23 patients with recurrence,9 had solitary tumor without lymphnode and distal metastases, for which the survival rates in 1,2 and 3 years were 100%,88.9%, and 35.6%, respectively, whereas in other recurrent patients the survival rates in 1,2 and 3 years were 21.4%, 10.7% and 0%, respectively(P< 0.01). Conclusions Ultrasound-guided microwave ablation in combination with TACE is effective for PLC patients with early stage. In recurrent PLC patients after ablation therapy with solitary tumor and no lymphnode and distal metastases the survival is significantly longer than that of the others.

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

Objective To investigate the feasibility and efifcacy of endoscopic submucosal dissection (ESD) for gastrointestinal neuroendocrine neoplasms (GI-NENs).Methods 52 patients with conifrmed histological diagnosis of GI-NENs performed ESD from January 2011 to December 2015 were included. The endoscopic morphology of tumor was summarized. Complete resection rate, complications, clinicopathological characteristics, and follow-up results were evaluated.Results There were 16 cases of stomach, 9 cases of colon and rectum 27 cases. Most of the lesions were submucosal uplift. A few of lesions looked like polyps. All the lesions were one-time whole diseased. 44 lesions were NET-G1, 8 lesions were NET-G2. Complete resection rate was 94.23%. 2 cases of rectal lesions infringemented intrinsic muscle layer, and got additional surgery. 1 case of rectal perforation, which was managed by endoscopic treatment and conservative treatment. All cases did not appear haemorrhage. During a mean follow-up period of 22.6 months, local recurrences occurred in 1 case of stomach, and treated with second line ESD. No cases lymph node and distant metastasis were found.Conclusion ESD appears to be a feasible, safe and effective treatment for GI-NENs with strict endoscopic treatment indications.

Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1∶ 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1∶ 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.

To establish a new immune assay for Penicillic Acid (PA) from Penicillium cyclopium, we studied the synthesis of conjugated complete antigens for penicillic acid. PA was conjugated to bovine serum album (BSA) and ovalbumin (OVA) by 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride (EDC). The artificial antigens PA-BSA and PA-OVA were identified by ultraviolet spectrometric scanning, SDS-PAGE and immunization. Results showed that the absorption peak of conjugation were different from that of the carrier protein alone and of the PA. The conjugated ratio of PA and BSA was 23.2:1 and that of PA and OVA was 10.4:1. Balb/c mice were immunized by the artificial antigen of PA-BSA, with PA-OVA as coating antigen. The average titer of antiserums was more than 12 800 by indirect ELISA. The obtained antigens offered a basis for developing immunoassay method.
Animals ; Antibodies ; blood ; Antigens ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Ethyldimethylaminopropyl Carbodiimide ; analogs & derivatives ; chemistry ; Immunization ; Mice ; Mice, Inbred BALB C ; Ovalbumin ; immunology ; Penicillic Acid ; immunology ; metabolism ; Penicillium ; immunology ; metabolism ; Serum Albumin, Bovine ; immunology