OBJECTIVETo investigate HLA genotyping by oligonucleotide arrays.

METHODUnsymmetrical PCR was used to amplify HLA-A gene exon 2, 3. The PCR products were used as templates for hybridization. The oligonucleotide probes were spotted on glass to make microarrays. High signal and specific probes were selected. The effects of the length and location of probes on hybridization signal were studied. A set of computer software was designed for scanning and genotype differentiation.

RESULTThe genotypes of 30 samples analyzed by microarray showed concordance to that by SBT and PCR-SSP.

CONCLUSIONHLA-A genotyping by oligonucleotide array is a good method with advantage of high speed, low cost and high flux.

Genotype ; HLA-A Antigens ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Oligonucleotide Probes ; Sensitivity and Specificity

HLA genes constitute a highly polymorphic multigene system. In the present study, HLA-B oligonucleotide chips were manufactured by using a set of sequence-specific oligonucleotide probes derived from polymorphic regions in exon 2 and exon 3 of HLA-B gene spotted by microarrayer onto the aldehyde modified glass slides. In addition, the sequenced HLA-B gene clones used as standard samples were amplified from exon 2 and exon 3 by PCR. Together with the correct hybridization and wash conditions, the PCR products were bound with the array probes on the chip, and the hybridization patterns were transformed to HLA-B genotypes. The results showed that the genotypes of standard samples by the HLA-B oligonucleotide chips were completely identical with the sequenced clones. In conclusion, the oligonucleotide chip method presented here for HLA-B genotyping is a rapid, accurate, sensitive and attractive high throughput biochemical way.
Genotype ; HLA-B Antigens ; classification ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; Sequence Analysis, DNA

Breast cancer has the highest incidence rate among all women's malignant tumors worldwide.Surgery,radiotherapy and chemotherapy are three major treatments,while most patients showed adverse effects or complications during or after the treatment,including lymphedema,gastrointestinal reactions and leukopenia,which cause severe impact on patients' recovery and quality of life.Moxibustion has been used and certified to alleviate adverse effects of surgery or chemoradiotherapy for breast cancer.We have summarized literatures in recent years and suggest more systematic research in the future for the underlying mechanism.

Clonal Evolution ; Humans ; Multiple Myeloma

This study was aimed to investigate the role of the delta-opioid receptor (DOR)-β-arrestin1-Bcl-2 signal transduction pathway in the pathogenesis of ulcerative colitis (UC) and the intervention effects of oxymatrine on UC. Forty Sprague-Dawley rats were divided into normal group, model group, oxymatrine-treated group and mesalazine-treated group (n=10 each) at random. The rat UC model was established by intra-colonic injection of trinitrobenzene sulfonic acid in the model group and two treatment groups. The rats in oxymatrine-treated group were subjected to intramuscular injection of oxymatrine [63 mg/(kg·day)] for 15 days, and those in mesalazine-treated group given mesalazine solution [0.5 g/(kg·day)] by gastric lavage for the same days. Animals in normal group and model group were administered 3 mL water by gastric lavage for 15 days. On the 16th day, after fasting for 24 h, the rats were sacrificed for the removal of colon tissues. The expression levels of DOR, β-arrestin1 and Bcl-2 were determined in colon tissues by immunohistochemistry and real-time quantitative polymerase chain reaction (RT-PCR), respectively. It was found that the expression levels of DOR, β-arrestin1 and Bcl-2 protein and mRNA were significantly increased in the model group as compared with the other groups (P<0.05). They were conspicuously decreased in both mesalazine-treated and oxymatrine-treated groups in contrast to the model group (P<0.05). No statistically significant difference was noted in these indices between mesalazine- and oxymatrinetreated groups (P>0.05). This study indicated that the DOR-β-arrestin1-Bcl-2 signal transduction pathway may participate in the pathogenesis of UC. Moreover, oxymatrine can attenuate the development of UC by regulating the DOR-β-arrestin1-Bcl-2 signal transduction pathway.
Alkaloids ; pharmacology ; Animals ; Anti-Arrhythmia Agents ; pharmacology ; Arrestins ; metabolism ; Colitis, Ulcerative ; metabolism ; pathology ; prevention & control ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Quinolizines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; metabolism ; Signal Transduction ; drug effects ; beta-Arrestins

In order to reveal variation and evolution of M genes of human avian H5N1 influenza strains, the M genes of human avian H5N1 strains in Guangdong were sequenced and the M genes of global strains were searched out from Internet. They were analyzed by DNAStar 5. 0 and their revolutionary speeds were studied by means of combining the epidemiological data. It was found that M1 genes of 53 H5N1 strains and M2 genes of 51 strains during 1997-2006 were homologously classified into two groups: the strains from Hong Kong during 1997 (G I) were one group and the strains from Hong Kong, Vietnam, Thailand, Indonesia, China mainland, Turkey, Iraq, Azerbaijan, Egypt during 2003-2006 (G II ) were the another group. There were 20 substitutions of amino acids in M1 gene of all strains (7.94%, 20/252), where there were 9 amino acids in strains during 2003-2006 differing from the strains in 1997, meanwhile there were 22 substitutions of amino acids in M2 gene of all strains (22.7%, 22/97), where there were 4 amino acids in strains during 2003-2006 differing from the strains in 1997. In the synonymous variation, Ks values in M1 were 26.8 x 10(-6)-42.6 x 10(-6) Nt/d, and Ka values 4.39 x 10(-6)-6.98 x 10(-6) Nt/d, where there was more rapid speed of synonymous substitution than that of replacement, which showed that there existed less human immunological pressure and negative selective pressure by biological test. Ks values in M2 were 13.1 x 10(-6)-23.4 x 10(-6) Nt/ d, and Ka values 9.1 x 10(-6)-16.2 x 10(-6) Nt/d; where the ratios of Ks to Ka was 1.0-1.6 times as there was the neutral selective pressure in TL-676-05 strain. There was an amino acid substitution of S224, N in M1 gene of strains during 2003-2006 and an increas in a glycoprotein domain NSS224-226. The secondary structure of M2 protein varied as the substitution of C50 F of eight strains from Indonesia in 2005. The strains G I did not reemerge after Hong Kong human avian H5N1 influenza event. An increase of a glycoprotein domain NSS224-226 in M1 protein during 2003-2006 might be related with virus pathogenicity. Human avian H5N1 influenza M gene evolved frequently in nature, which might have an impact on its capacity of human-to-human transmission.
Amino Acid Sequence ; China ; Genetic Variation ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; Molecular Sequence Data ; Phylogeny ; Viral Matrix Proteins ; chemistry ; genetics

Objective To evaluate the effcacy and side effects of [~(90)Y-1,4,7,10-tetraazacyclododecane-N,N',N",N'"-tetraacetic acid (DOTA)~0,Tyr3]octreotate (~(90)Y-DOTATATE) combined with lysine as a renal protective agent.Methods Twenty-five patients with advanced neuroendocrine tumors were confirmed as somatostatin receptor subtype-2 (SSTR2)-positive by somatostatin receptor scintigraphy (SRS).Each patient received 1-5 cycles of treatment and the interval between two cycles of treatment was 6-9 weeks.~(90)Y-DOTATATE was administered intravenously within 30 min.Lysine was injected before and after the administration of ~(90)Y-DOTATATE.After each treatment cycle.the side effects were assessed according to National Cancer Institute Grading Criteria(NCIGC).The etticacy was evaluated by the WHO criteria 8 weeks after the last treatment.Results Pattial remission was found in 1 patient (4%).minor response in 3 patients(12%),stable disease in 16 patients (64%)and tumor progression in 5 patients (20%).Two patients suffered from renal functional injuries and 3 patients developed leukocytopenia.Three patients showed nausea while another 3 patients presented vomiting.Conclusions ~(90)Y-DOTATATE in association with lysine may be a promising treatment method for the patients with metastatic and inoperable neuroendocrine tumors.More research work may be directed to reduce renal injury.

Objective:To observe the clinical effect of acupuncture for persistent allergic rhinitis (PAR).Methods:A total of 154 PAR patients were randomized into a treatment group and a control group,78 cases in the treatment group received acupuncture at meridian points,while 76 cases in the control group received acupuncture at non-meridian points.The treatment was done every other day,3 times a week for a total 4 weeks.The total nasal symptom score (TNSS),the total non-nasal symptom score (TNNSS) and the rhinoconjunctivitis quality of life questionnaire (RQLQ) were measured before the treatment and after 2 weeks and 4 weeks of treatment,as well as 1 month and 3 months after the treatment to compare the clinical effect between the two groups.Results:After 4 weeks of treatment,the total effective rate in the treatment group was 92.3％,versus 76.3％ in the control group,showing a statistically significant difference (P＜0.05).Intra-group comparisons of TNSS,TNNSS and RQLQ scores after 4 weeks of treatment showed statistically significant differences (all P＜0.05).Between-group comparisons of TNSS,TNNSS and RQLQ score after 4 weeks of treatment and in follow-up visits showed statistically significant differences (all P＜0.05).Conclusion:Acupuncture is effective for PAR,and acupuncture at meridian points can produce a better effect than acupuncture at non-meridian points.

Objective: To explore the changes of inflammatory factors and related factors in the population with overweight combining abdominal obesity and high-normal blood pressure (BP). Method: Our research included in 2 groups: Group A: n=189 subjects with high-normal BP, overweight and abdominal obesity, their BMI ≥ 24 kg/m2, waist circumference (WC) ≥ 90 cm in male, WC ≥ 85 cm in female, SBP(120-139) mmHg or DBP (80-89) mmHg; Group B, n=87 healthy subjects with matched age, BMI < 24 kg/m2, BP < 120/80 mmHg as normal control. Blood lipids and other biochemical parameters were examined; serum levels of intercellular adhesion molecule-1 (ICMA1), monocyte chemoattractant protein-1 (MCP1), chemokines-1 (CXCL-1), CXCL-2 and oxidized low density lipoprotein (oxLDL) were measured by ELISA. Results: Compared with Group B, Group A had increased TG, fasting blood glucose and non-HDL-C, all P<0.05; elevated serum levels of ICMA1 and MCP1, both P<0.05. Correlation analysis indicated that in Group A, ICMA1 was positively related to BMI, SBP, LDL-C and negatively related to age, which had gender difference; MCP1 was positively related to WC, SBP, LDL-C, non-HDL-C and negatively related to HDL-C, which also had gender difference; oxLDL was positively related to SBP, LDL-C; no evidence showed that CXCL-1 and CXCL-2 were related to obesity, BP and metabolic parameters; in Group B, no evidence showed that inflammatory factors were related to the other parameters. Linear regression analysis for inflammatory parameters found that after excluding other factors, in Group A, ICMA1 was positively related to BMI (t=2.901, P=0.005); in male gender, MCP1 was positively related to SBP (t=5.076, P=0.000), negatively related to DBP (t=-3.369, P=0.001). oxLDL was positively related to age (t=2.168, P=0.032) and LDL-C (t=2.146, P=0.034); CXCL-1 was negatively related to HDL-C (t=-2.013, P=0.047). Conclusion: The subjects with overweight abdominal obesity and high-normal BP were usually having abnormal metabolism of glucose and lipids, elevated serum levels of inflammatory parameters, blood levels of inflammatory factors were increasing with elevated BMI and SBP accordingly which implied the association with critical range of BP.

OBJECTIVETo compare the clinical effectiveness of percutaneous US- or CT-guided drainage and laparotomy for patients with infective pancreatic necrosis.

METHODSData of 90 infective pancreatic necrosis patients admitted from January 2008 to December 2009 were included. They were divided into two groups by the different treatment choices. Twenty-seven patients in the percutaneous group received percutaneous US- or CT-guided drainage as first choice. After that a sump suction apparatus was applied for controlled drainage. If no improvement was achieved after 3 days, they would received operation soon. While patients in the laparotomy group received surgical drainage at the time when the diagnosis of infected pancreatic necrosis was confirmed. Continuous drainage was also applied for these ones.

RESULTSThe percutaneous group had a significant low rate of reoperation (7.1% vs. 14.3%, P < 0.05) and postoperative residual abscesses (7.1% vs. 28.6%, P < 0.05). Furthermore, 48.1% of patients in percutaneous group successfully avoid laparotomy. In the regard of complications, the percutaneous group presented lower incidence of both single organ dysfunction (7.4% vs. 28.6%, P < 0.05), intestinal fistula (7.4% vs. 27.0%, P < 0.05) and long-term complications (3.7% vs. 22.2%, P < 0.05). In addition, the percutaneous group costed less medical resources as evidenced by shorter ICU duration (21.2 ± 9.7 vs. 28.7 ± 12.1, P < 0.01), shorter hospital duration (48.2 ± 12.5 vs. 59.6 ± 17.5, P < 0.05) and less expenditure (191 762 ± 5892 vs. 341 689 ± 10 854, P < 0.05).

CONCLUSIONSPercutaneous drainage can effectively lower the surgical rates and the rates of complications and reoperations in patients with infective pancreatic necrosis. Besides that, it could also reduce the cost of medical resources.

Adult ; Drainage ; methods ; Female ; Humans ; Laparotomy ; Male ; Middle Aged ; Pancreatitis, Acute Necrotizing ; surgery ; Retrospective Studies ; Treatment Outcome