M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

OBJECTIVETo seek the effective treatment for peripheral neuropathy induced by chemotherapeutic drugs.

METHODSSixty-four cases of peripheral neuropathy induced by Paclitaxel or Oxaliplatin were randomly divided into an acupuncture group and a medication group, 32 cases in each group. The acupuncture group was treated with therapeutic principle of dredging meridians and collaterals, tonifying qi and eliminating blood stasis, supplementing liver and kidney, nourishing blood and tendon. Hegu (LI 4), Taichong (LR 3), Zusanli (ST 36), Qihai (CV 6) and Quchi (LI 11) etc. were selected. The medication group was treated with intramuscular injection of Cobamamide. The neurotoxicity of two groups was compared with questionnaire of peripheral neuropathy induced by chemotherapeutic drugs before and after treatment.

RESULTSThe total effective rate for sensory nerve disorder of acupuncture group was 66.7% (20/30), which was superior to that of 40.0% (12/30) in medication group (P < 0.05).

CONCLUSIONAcupuncture is more effective than Cobamamide for treatment of peripheral neuropathy induced by chemotherapeutic drugs, especially for moderate and severe sensory nerve disorder induced by paclitaxel.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Drug-Related Side Effects and Adverse Reactions ; Female ; Humans ; Male ; Middle Aged ; Organoplatinum Compounds ; adverse effects ; Paclitaxel ; adverse effects ; Peripheral Nervous System Diseases ; chemically induced ; therapy

Nine compounds were isolated from the n-butanol fraction of 95% ethanol extract of the fruit of Alpinia oxyphylla Miq. with a combination of various chromatographic approaches, including MDS resin, silica gel, reverse phase C18 and preparative HPLC. On the basis of spectroscopic data analysis, they were elucidated as (1R, 4R, 10R)-1β, 4α-dihydroxy-11, 12, 13-trinor-5, 6-eudesmen-7-one (1), 1β, 4β-dihydroxy-11, 12, 13-trinor-8, 9-eudesmen-7-one (2), oxyphyllenone A (3), oxyphyllenone B (4), rhamnocitrin (5), staphylionoside D (6), benzyl-1-O-β-D-glucopyranoside (7), 2-O-β-D-glucopyranosyl-(1S)-phenylethylene glycol (8), and (S)-1-phenylethyl-β-D-glucopyranoside (9). Among them, compound 1 is a new sesquiterpene, named as oxyphyllenone C; compounds 8 and 9 are new natural products; compounds 2 and 6 were isolated from the genus Alpinia for the first time, and compound 7 was isolated from A. oxyphylla for the first time.
1-Butanol ; Alpinia ; chemistry ; Chromatography, High Pressure Liquid ; Fruit ; chemistry ; Phytochemicals ; chemistry ; isolation & purification ; Sesquiterpenes ; chemistry ; isolation & purification

Objective: To assess the effect and mechanism of compound Xiansu capsule (仙酥胶囊, XSC) combined with chemotherapy in treating gastric carcinoma of mid-late stage. Methods: Sixty-one patients of the test group were treated by XSC combined with chemotherapy and 30 patients of the control group treated with chemotherapy alone. The effect of treatment and cell mediated immunity of patients were observed. Results: The effective rate of the test group and the control group was 32.8% and 13.3% respectively (P<0.05), the toxic reaction occurrence caused by chemotherapy was less in the former than that in the latter group (P<0.01). The CD8 level of patients in the test group decreased, and CD4/CD8 level was raised obviously, which suggested that XSC had immuno-regulating effect on T-cell. Conclusion: XSC could enhance the efficacy and reduce the toxic and side-effect of chemotherapy. To regulate the cell mediated immunity of patients is possibly its mechanism.

Traditional Chinese medicine is a treasure of Chinese culture, absorbing the wisdom of the Chinese people. Continuous application of new technologies makes traditional Chinese medicine research advance with the times. After several years of development, high-throughput transcriptome study has become a mature research tool in biology. This paper reviewed the advances in medicine transcriptome study, and compared two sequencing platforms, Roche's GS FLX platform and Illumina's HiSeq 2000 platform. Moreover, this paper introduced medicine transcriptome analysis process, with Panax quinquefolius and Lonicera japonica for examples, showing the characteristics of traditional Chinese medicine transcriptome studies. High-throughput transcriptome studies facilitate traditional Chinese medicine research with overall understand of functional genes, give clear elucidation of metabolic pathways, lay molecular foundation for the traditional Chinese medicine research and offer modern interpretation for traditional Chinese medicine theory. However, the current study faces several difficulties, including weak molecular basis, high sequencing cost and staff shortages in data anaysis. In the future, with the development in sequencing technology, the combination of transcriptome and other genomics, such as proteome and metabolome, will lay a solid foundation for the new high-throughput screening and developing model for the traditional Chinese medicine industry.
Biomedical Research ; methods ; trends ; Forecasting ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Plant ; Humans ; Lonicera ; genetics ; Medicine, Chinese Traditional ; methods ; trends ; Panax ; genetics ; Phytotherapy ; methods ; trends ; Transcriptome ; genetics

OBJECTIVETo discuss the surgical option and the treatment of complications of pancreatic cystic tumors.

METHODSFrom January 1997 to December 2009, 32 patients with pancreatic cystic tumors in our center were reviewed retrospectively. There were 6 male and 26 female, aging from 24 to 76 years. Of the 32 patients, 16 patients had serous cystadenoma, 9 patients had mucinous cystadenoma; 1 patients had mucinous cystadenocarcinoma; 4 patients had intraductal papillary mucinous neoplasms and 3 patients had pancreatic solid pseudopapillary neoplasms. Tumor located in pancreatic head in 12 patients and in pancreatic body and tail in 20 patients.

RESULTSAll patients received surgical treatment and there was no perioperative death. Pancreato-duodenectomy was performed in 10 patients, duodenum-preserving pancreatic head resection in 1 patient, distal pancreactomy in 13 patients, including laparoscopic distal pancreactomy in 2 patients, pancreatic tumor resection in 3 patients, middle segmental resection in 4 patients; 1 patients with mucinous cystadenocarcinoma received palliative surgery. Complication included gastroparesis in 3 patients and pancreatic fistula in 5 patients, and all recovered by conservative treatment. These 29 patients were followed up 4 - 120 months, 3 patients died from tumor metastasis or other disease within 4 to 34 months after surgery. Others were alive and there was no tumor recurrence or metastasis.

CONCLUSIONSCT scan should be the first choice of non-invasive examination for cystic pancreatic diagnosis. Positive and timely operation should be performed in the patient with cystic pancreatic tumor, and it acts as a cancer preventive treatment. The selection of surgical approach should be individualized, the principal of damage control surgery should be followed. Complications such as gastroparesis and pancreatic fistula should be paid more attention.

Adult ; Aged ; Cystadenoma, Mucinous ; diagnosis ; surgery ; Cystadenoma, Serous ; diagnosis ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; surgery ; Retrospective Studies

OBJECTIVETo detect the IgH-MMSET fusion gene resulted from t (4;14) translocation in multiple myeloma and illuminate its significance.

METHODSIgH-MMSET fusion gene was detected in bone marrow specimens of 25 multiple myeloma (MM) patients and MM cell line NCI-H929 using reverse-transcription PCR (RT-PCR) assay followed by nested PCR to increase the sensitivity. The purified PCR products were cloned into pGEM-T vector and then sequenced using M13 forward primers. The fragment sequences were compared with that in GenBank to find matched sequences.

RESULTSOnly a 438 base pair long fragment was obtained after RT-PCR assay and was confirmed by sequencing to be a fusion gene product of IgH gene and MMSET gene in MM cell line NCI-H929. The breakpoints were located within the C micro region of IgH gene on chromosome 14 and intron 3 of MMSET gene on chromosome 4. IgH-MMSET hybrid transcripts were detected in 3 of 25 MM patients through nested PCR assay. The amplified fragments of the 3 patients were 237 base pairs (bp), 239 bp and 239 bp in length, respectively. The breakpoints on chromosome 4 were identical to that of NCI-H929 cell.

CONCLUSIONSThe formation of IgH-MMSET fusion gene is resulted from t (4;14) translocation in MM. The incidence rate is 12.0%. The presence of IgH-MMSET fusion gene may predict poor prognosis.

Adult ; Aged ; Base Sequence ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 4 ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; Oncogene Proteins, Fusion ; genetics ; Protein-Tyrosine Kinases ; Receptor, Fibroblast Growth Factor, Type 3 ; Receptors, Fibroblast Growth Factor ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Translocation, Genetic

OBJECTIVETo evaluate the effect of polymorphism at the -238 and -308 position of the TNF-alpha promotor region on the clinical outcome of thalidomide (Thal)-based regimens for the treatment of multiple myeloma (MM).

METHODSThe polymorphism at the -238 and -308 position of the TNF-alpha promotor region of 168 MM patients treated with Thal-based regimens were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Genotypes were tested for association with overall response by logistic regression, and survival was evaluated by univariate and multivariate analysis.

RESULTSIn TNF-alpha -238 position, 11 (6.5%) patients had GA genotype and 1 (0.6%) AA genotype. In TNF-alpha -308 position, 19 (11.3%) had GA genotype and 1 (0.6%) AA genotype. In univariate analysis, the TNF-alpha -238 GA + AA genotypes were associated with a significantly prolonged progression free survival (PFS) (P = 0.017), and a better overall survival (OS) (P = 0.150). Multivariate COX regression analysis showed that TNF-alpha -238 polymorphic status was an independent prognostic factor for prolonged PFS (P = 0.049).

CONCLUSIONThe TNF-alpha -238 polymorphic status is associated with a favorable clinical outcome in MM patients treated with thalidomide-based regimen. The polymorphism status of TNF-alpha gene might be of promise for developing a more informative stratification system for MM.

Adult ; Aged ; Aged, 80 and over ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; genetics ; Polymorphism, Genetic ; Prognosis ; Promoter Regions, Genetic ; Thalidomide ; therapeutic use ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; genetics

OBJECTIVETo develop an HPLC method for determining oxyphyllenodiol A (1) and teuhetenone A (2) contained in Alpinia oxyphylla and to compare the contents of the two components contained in medicinal materials and prepared herbal medicines in pieces sold in the market and different fractions.

METHODHPLC and Waters sunfire C18 column (4. 6 mm x 250 mm, 5 microm) were adopted for gradient elute with the mobile phase of acetonitrile and water. The flow rate was 1.0 mL x min(-1) and the detection wavelength was 250 nm.

RESULT1 and 2 showed a good linear relationship within the range of 0.1296 - 0.8640 microg and 0.1635 - 1.0900 microg respectively, with the average recoveries of 99.08% and 97.80%. Their content ranges were 0.0059% - 0.0149% and 0.0080% - 0.0164% in different samples. The mean value of 1 and 2 were 0.0085% and 0.0104% in the whole fruits, and 0.0137% and 0.0157% in the seeds. They were undetected in the nutshells.

CONCLUSIONThe method is so precise, accurate and highly reproducible that it can be used to determine the contents of oxyphyllenodiol A and teuhetenone A in A. oxyphylla. The contents of the two components are mainly extracted from the seeds, with certain difference among different samples. There are a higher contents and no significant difference in the salted and raw seeds.

Alpinia ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Sesquiterpenes ; analysis ; isolation & purification

OBJECTIVETo study the relationship between STXBP1 gene mutations and refractory seizures with unknown causes in newborns.

METHODSThe coding region of STXBP1 gene was detected using direct Sanger sequencing in 11 newborns with refractory seizures of unknown causes.

RESULTSSTXBP1 gene mutation was found in 1 out of 11 patients. It was a missense mutation: c.1439C>T (p.P480L).

CONCLUSIONSSTXBP1 gene mutation can be found in neonatal refractory seizures of unknown causes, suggesting a new approach of further research of this disease.

Humans ; Infant, Newborn ; Munc18 Proteins ; genetics ; Mutation ; Seizures ; genetics