Objective:To optimize the processing technology of prepared Morinda officinalis based on nystose content. Methods:Orthogonal experiments and the nystose determination method in Chinese Pharmacopoeia (2010 edition) were used to optimize the pro-cessing technology. A Welch AQ-C18(250 mm ×4.6 mm,5 μm)chromatographic column was applied with methanol-water(3∶97)as the mobile phase, the column temperature was at 25℃, the flow rate was 0. 8 ml·min-1 , the injection volume was 10μl, and the drift tube temperature was at 40℃ with low temperature atomization. Results:The linearity of nystose was good within the range of 0. 214 8-0. 644 4 mg·ml-1 and the recovery was 99. 5%(RSD=3. 5%,n=6). Conclusion: On the basis of nystose content determination, the optimal processing technology of prepared Morinda officinalis is as follows:liquorice of 4%, stir fry time of 15min and the boiling pot temperature of 200℃.

Adolescent ; Adult ; Blood Transfusion ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Pesticides ; poisoning ; Poisoning ; therapy ; Treatment Outcome

Objective To investigate the effect of trichostatin A (TSA) and 5-azacitidine (5-AzaC) on pancreatic β-cells impaired by cytokine,via measuring the proliferation,apoptosis,and function of pancreatic β-cells.Methods RIN-m5f was impaired by interleukin-1β and interferon-γin vitro,and treated with TSA and 5-AzaC.Experiment groups included blank control group,cytokine induction group,0.05/0.10 μmoL/L TSA group,0.63/1.25 μmoL/L 5-AzaC group,and0.10 μmol/L TSA plus 1.25 μmol/L 5-AzaC group.The viability of RIN-m5f cells was detected by MTT assay.Apoptotic rate was determined by Annexin V-fluorescein isothiocyanate (FITC) /propidium iodide flow cytometry.Insulin secretion was measured by enzyme-linked immunosorbent assay.Results The viability of RIN-m5f cells in 0.05/0.10 μmoL/L TSA group,0.63/1.25 μmol/L 5-AzaC group,and 5-AzaC plus TSA group was 70.1％/79.2 ％,67.3 ％/82.9 ％,and 89.1％ respectively,being higher than that in the cytokine group (33.9％,P＜0.05) ; the apoptosis rate was 10.3％/10.5％,7.9％/9.6％,and 8.2％,being lower than that in the cytokine group (16.6％,P＜0.05) ; the capacity of glucose-stimulated insulin secretion of all the treated groups was higher than that in the cytokine group (P＜0.05).Conclusion TSA and 5-AzaC might promote the proliferation of pancreatic β-cells impaired by cytokines,inhibit its apoptosis and recover its insulin secretion.

Objective To study histone acetylation patterns in peripheral boiood mononuclear cells (PBMC) from type 2 diabetic patients PBMC form 12type 2 diabetic patients and 12 healthy contrcl subiects were collecyed The global histone H3/H4 acetylation was determined by assay kit The differetial expression of the histone acetyltransferases and histone deacetylases were measured by real time PCR.Results increased H3/H4 acetylation was observed in PBMC of type 2 diabetic patients compared with control subjects (H3:0.134±0.035vs0.181±0.032,p<0.05;H4:0.266±0.070VS0.324±0.062,P<0.01).the mRNA levels of p300,CREBBP,HDAC2,HDAC7,and STRTL in patients PBMC compared with healthy control subieects were 3.11±0.38,2.78±0.45,3.55±0.30,0.77±0.37 and 0.40±0.25 folds respectively conclusion histone acetylation appears abnormal in PBMC of type 2 diabetic patients.

Objective To explore the characteristics of proinsulin secretion in latent autoimmune diabetes in adults (LADA). Methods Fasting and 2 h sera in oral glucose tolerance test from 36 LADA patients, 37 type 2 diabetic patients and 43 healthy controls were collected to test glucose, proinsulin (PI) and C-peptide (CP) by radioimmune assay. Glutamie acid decarboxylase antibodies (GAD-Ab) were determined by radioligand assay.Results (1) Fasting proinsulin (FPI) and 2 h proinsulin (PPI) level in LADA patients were lower than those in type 2 diabetic patients (P<0.05), being both significantly inereasad compared with healthy controls (P<0.05 or P<0.01); The ratios of FPI/FCP and PPI/PCP (%) in LADA were beth significantly higer than those of type 2 diabetes mellitus and controls (P<0.05 or P<0.01); (2) LADA type-1 (GAD-Abe>0.3) patients showed lower PI levels(P<0.05 or P<0.01) and higher PI/CP ratio (all P<0.05) than LADA type-2 (0.05≤GAD-Ab<0.3); Meanwhile, there were no significant differences in above parameters between LADA type-2 and type 2 diabetes meUitus (P>0.05). (3) GAD-Ab index was negatively correlated with FPI and PPI in LADA group (r=-0.236 and-0.268, both P<0.05), and positively correlated with PPI/PCP (r=0.254, P=0.030).Meanwhile BMI was positively correlated with FPI, PPI and PI/CP in type 2 diabetes mellitus (all P<0.01). No factor entered the multiple regression analysis for predieting the hyperproinsulinemia and dispropriately elevated proinsulin levels in LADA patients. (4) According to the 99.5 th percentile of proinsulinemia in the healthy controls, which is defined as the cutoff point dispropriately elevated proinsulin levels, the proportion of subjects with fasting dispropriately elevated proinsulin levels (FPI/FCP) were 77.8%, 62.2% and 2.3% in LADA, type 2 diabetes meUitus and controls respectively, and PPI/PCP 83.3%, 51.4% and 2.3% respectively. Conclusion LADA patients, as well as type 2 diabetic patients, all showed hyperproinsulinemia and disproportionately elevated proiasulin levels that were one of characteristics of defective β-cell function. Moreover, disproportionately elevated nproinsulin level is more evident in LADA patients than that in type 2 diabetics and this may be related to humoral immunity.

BACKGROUNDMutations in mitotic checkpoint genes have been detected in several human cancers, which exhibit chromosome instability. We wanted to know whether mutation of hBub1 could occur in transformed human embryo lung fibroblasts (HELF) cells induced by a chemical carcinogen.

METHODSHELF cells were transformed by N-methyl-N'-nitro-N-nitrosoguaridine (MNNG), and three flasks of transformed HELF cells (named as T1, T2, and T3) were selected as amplifiers, and mutations of hBub1 in these transformed cells were analyzed by PCR-SSCP and sequencing.

RESULTSIt was found that any one of three transformed cell lines exhibited aneuploidy with a low mitotic checkpoint function. Subsequent PCR-SSCP and sequence analysis showed an AGT to CGT or ATT mutation at codon 80 in hBub1 gene in T1 cells with a resultant change in amino acid sequence.

CONCLUSIONOur study demonstrated that the mitotic checkpoint genes could be targets of MNNG.

Cell Line, Transformed ; Chromosome Aberrations ; Down-Regulation ; Fibroblasts ; drug effects ; Humans ; Lung ; cytology ; Methylnitronitrosoguanidine ; toxicity ; Mitosis ; drug effects ; Mutation ; Protein Kinases ; genetics ; Protein-Serine-Threonine Kinases

Objective： The current study was designed to investigate the effect of heat shock protein 70(HSP70) on stability of mutant p53 protein. Methods： Retroviral recombinant expressing antisense HSP70 RNA was constructed and transfected into MCF7/Adr breast cancer cells. The existence of foreign DNA was identified by PCR method and the HSP70 protein level was determined by Western blot analysis. The half-life of mutant p53 protein(mtp53) was measured by p53 stability assay. Results： The stable expressing strain(MAp70) from transfected cells was obtained through G418 selection. The foreign DNA in transfectant cells were confirmed by PCR, and the repression rate of HSP70 protein was 42％ . The half-life of mutant p53 in MAp70 cell was 12 hours, significantly lower than that of the control cells. Conclusion： Antisense HSP70 RNA can decrease the HSP70 protein level and significantly increase instability of mutant p53 protein in MCF7/Adr breast cancer cell.

OBJECTIVETo set up a technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene, and to evaluate the possibility of using this technique for preimplantation genetic diagnosis(PGD) of deleted Duchenne muscular dystrophy (DMD) with family history.

METHODSFifty single lymphocytes of a normal male and fifty of a normal female were obtained for detecting dystrophin gene(exon 51, exon 19, exon 48) and SRY gene by 3-plex nested PCR.

RESULTSIn the group of exon 51/exon 19/SRY, the amplification rates of exon 51, exon 19 and SRY in male were 96%, 94% and 94%; the amplification rates of exon 51 and exon19 in female were 94% and 94%, respectively. In the exon 48/exon 19/SRY group, the amplification rates of exon 48, exon 19 and SRY in male were 92%, 90% and 94%, the amplification rates of exon 48, exon 19 in female were 94% and 92%, respectively.

CONCLUSIONThe technique of single lymphocytes 3-plex nested PCR for dystrophin and SRY gene established in this study is highly sensitive, specific and reliable, and is suitable for PGD of deleted DMD with family history.

Dystrophin ; genetics ; Exons ; genetics ; Female ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Preimplantation Diagnosis ; methods ; Reproducibility of Results ; Sequence Deletion ; Sex Determination Processes

We report here a case of pentastomiasis infection in a 3-year-old girl who had high fever, abdominal pain, abdominal tension and anemia. Ultrasound scanning of the abdomen revealed disseminated hyperechoic nodules in the liver and a small amount of ascites. Abdominal MRI showed marked hepatomegaly with disseminated miliary nodules of high signal intensity throughout the hepatic parenchyma on T2-weighted images; retroperitoneal lymphadenopathy and disseminated miliary nodules on the peritoneum were also noted. Chest CT showed scattered small hyperdense nodules on both sides of the lungs. The laparoscopy demonstrated diffuse white nodules on the liver surface and the peritoneum. After the small intestinal wall and peritoneal biopsy, histological examination revealed parenchymal tubercles containing several larvae of pentastomids and a large amount of inflammatory cell infiltration around them. The pathological diagnosis was parasitic granuloma from pentastomiasis infection.
Abdomen, Acute/*parasitology ; Animals ; Biopsy ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging/methods ; Parasitic Diseases/*diagnosis ; *Pentastomida ; Tomography, X-Ray Computed/methods

OBJECTIVETo evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis (PGD) in couples at risk of having child with beta-thalassemia.

METHODSSingle cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia.

RESULTSIn six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out(ADO) rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination.

CONCLUSIONThis is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene(HumTHO1) could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.

Adult ; Female ; Humans ; Male ; Polymerase Chain Reaction ; Pregnancy ; Preimplantation Diagnosis ; methods ; beta-Globins ; genetics ; beta-Thalassemia ; diagnosis ; genetics