OBJECTIVE To detect 315 codon of mutation site in katG of isoniazid(INH)-resistant Mycobacterium tyberculosis(MTB) by stem-ring molecular probe quickly and detect out the fluorescence sign of hybridization between amplified products of katG 315 codon and probe in liquid by fluorescence spectrophotometer.The results were confirmed by sequencing.METHODS The software,Beacon designer,was used to design the katG 315 codon stem-ring molecular probe and the amplification system,and the relationship between the way and sequencing of the amplification products were compared.RESULTS The difference between PCR products from standard strain and INH-resistant one was significant in detecting the fluorescent light by use of fluorescence spectrophotometer.We detected fluorescent light signal between the 16 INH resistant strains and 10 H37RV standard strains.The resistant rate to INH detected was about 44%,and the rate of coincidence was about 97.5%.CONCLUSIONS The stem-ring molecular probe technology show high sensitivety in detecting mutation site of nucleic acid.The rate of coincidence is good between fluorescence spectrophotometer way and sequencing.

Objective: To investigate the killing activity of cytokine-induced killer (CIK) cells after incubation with autologous tumor cell lysate-pulsed dendritic cells (Ag-DC) and to evaluate the immune functions of patients, the clinical efficacy and side effect of Ag-DC in combination with CIK on hepatocellular carcinoma. Methods: Peripheral blood mononuclear cells were isolated from 24 patients with hepatocellular carcinoma, and cultured with granulocyte-macrophage colony-stimulating factor and interleukin-4 to produce dendritic cells (DCs). The DCs were pulsed with autologous tumor cell lysate. T lymphocytes from PBMC were cultured with interferon-Y (IFN-γ), IL-2, CD3-moAb, and IL-1α to prepare CIK. The killing effect of CIK on SMMC-7721 was investigated after CIK was incubated with Ag-DC. After immunotherapy with Ag-DC and CIK, immunolog-ic and clinical responses of the 24 patients were evaluated. Results: The killing effect of CIK was remarkably improved after CIK was incubated with Ag-DC. The immunotherapy with DC and CIK alleviated symptoms and improved the immune functions of the patients. Except for transient fever and chill, no remarkable adverse events were observed. Conclusion: Ag-DC in combination with CIK shows short-term efficacy on hepatocellular carcinoma through inducing specific anti-tumor immunity and can be an effective adjuvant therapy for hepatocellular carcinoma.

Objective To review the three-dimensional characteristics and configuration of cranial base arteries of patients with intracranial berry aneurysms. Methods All the 70 patients with intracranial berry aneurysms (83 cases)were admitted from January to December in 2007. Their images of spiral computer angiography (CT) were presented and analyzed retrospectively. The site, size and figures of aneurysms were recorded, especially the variations of cranial base arteries were analyzed. Results Cerebral anterior communicating artery (ACoA) was the frequent site where aneurysms located. Incidence of ACoA aneurysms was 43.37%(36/83), that of posterior communicating artery (PCoA) was 28.92%(24/83), that of internal carotid artery was 9.64% (8/83), that of middle cerebral artery was 6.02% (5/83), that of A1-A3 was 3.61% (3/83), that of basilar artery was 3.61% (3/83), that of posterior cerebral artery was 2.41% (2/83), that of posterior inferior cerebellar artery was 1.20% (1/83),that of anterior choroidal artery was 1.20% (1/83).The shape of many berry aneurysms was regular. Mutational rate of cranial base arteries among aneurysms were 56.63% (47/83) and ACoA ancurysms with A1 dysplasia rate was 72.22% (26/36). Conclusions The solid shape and vascular variation of intracranial berry aneuryams can be optimally identified by spiral computer angiography. The occurrence of berry aneurysms might be associated with variation of cranial base arteries.

OBJECTIVETo study the therapeutic effects of Rhizoma Drynariae and estrogen on osteoporosis in ovariectomized (OVX) rats.

METHODSFifty-five female rats were randomly divided into 5 groups, the normal control (normal),the sham operated (sham), the model, the estrogen, and the Rhizoma drynariae (RD) groups; ovariectomized rats were used as postmenopausal osteoporosis model. The changes of morphology and dynamic parameters in different groups were determined by bone histomorphometry.

RESULTSCompared with the model group, the trabecular volume (TBV/TTV) , trabecular thickness (MTPT) and density (MTPD) in the other four groups were significantly increased, while the trabecular template spacing (MTPS) and the ratio of trabecular surface to trabecular volume (TBS/TBV) significantly decreased (P <0. 05); and the osteoid surface (TOS), single label surface [Sfract (s) ] ,double label surface [Sfract (d) ] and bone formation rate (Svf) also decreased,while osteoid maturation period (OMP) increased in the latter four groups. No significant difference of cortical width (MCW) was found between these 5 groups. Compared with the normal and sham groups, TOS, Sfract ( s) , Sfract ( d) , Svf in the estrogen and RD groups increased significantly, while OMP decreased; no significant difference was found in other parameters.

CONCLUSIONRhizoma Drynariae has the similar effect with estrogen in maintaining normal trabecular structure and connection by inhibiting the increased bone turnover of postmenopausal osteoporosis.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Estrogens ; therapeutic use ; Female ; Humans ; Osteoporosis ; drug therapy ; Ovariectomy ; Polypodiaceae ; Postmenopause ; Rats

ObjectiveTo conduct a molecular epidemiological study on human metapneumovirus (hMPV) among pediatric patients in Guangzhou. MethodsA total of 1 840 clinical specimens were obtained from pediatric patients with respiratory infections in Guangzhou Women and Children' s Medical Center in 2010.hMPV was detected by real-time TaqMan RT-PCR in clinical specimens.F gene was amplified and the PCR-products were directly sequenced. ResultsIn 1 840 clinical specimens, 66 werehMPV-positive with a positive rate of 3.59％. hMPV was detected in all specimens except those collected in September and October, and the highest positive hMPV rate occurred in April (6.09％). The F genes of 3 randomly selected strains and hMPVgz01 ( isolated in 2008) were compared with subgroups A1, A2, B1,B2 and C, and the highest homology was with BJ1887 strain of genotype A2b (97％). The F genes of the randomly selected strains and hMPVgz01 were 99％ identical to each other. Sequences and phylogenetics analysis revealed that the epidemical strain in Guangzhou belonged to genotype A2b. ConclusionhMPV is prevalent in spring and summer among children in Guangzhou, and A2b is the predominant genotype.

Objective To detect the blood parameter, blood biochemical and electrolyte indices of the F1 generation of Rongshui miniature pig ( RMP) .Methods The blood of 43 female and 42 male RMPs of 4 th month old, and 36 RMPs of 12th month old ( half male and female) were extracted from jugular vein.And the blood parameter, blood biochemical and electrolyte indices were detected by blood analyzer and automatic biochemical analyzer.Results In the same month-old RMP, no significant difference between male and female were found in most indices of blood parameter, blood biochemical and electrolyte indices.On the other hand, many indices were difference between 4th month old and 12th month old RMPs of same gender.Compared with the 4th month old RMP, the 12th month old RMP decreased significantly in WBC and PLT, increased in HGB ( P <0.05 ) while RBC was the same ( P >0.05 ) .Serum ALT, AST, ALP, CK (male), LDH(male), A/G, BUN, GLU (female), CHOL (male) and K+decreased significantly (P <0.05), while serum TP, TBIL, CR and Ca2+increased significantly (P <0.05),but serum CHOL, TG, HDL-C and LDL-C were not different.86.4%(19/22) biochemical and electrolyte indices in RMP were in/or close to the range of normal value of human.Conclusion Most of the blood parameter, blood biochemical and electrolyte indices of RMP were close to human’ s normal value.

OBJECTIVETo investigate the secretion patterns of glucose-dependent insulinotropic polypeptide (GIP) after different dietary loads in subjects with normal glucose tolerance (NGT) and their relation to insulin secretion and plasma glucose levels.

METHODSFourteen subjects with normal glucose tolerance underwent 75 g glucose tolerance test(OGTT) followed by mixed meal tolerance test(MMT) one week later. Blood glucose, insulin, and GIP were measured in the fasting state and at 0, 15, 30, 60, 90 and 120 min after glucose load or mixed meal load.

RESULTSThe first peak value of GIP after glucose load occurred at 15 min (45.09∓4.67 pmol/L). After a brief decline, GIP continued to increase till reaching 59.66∓11.73 pmol/L at 120 min after the load. After the mixed meal load, GIP secretion presented with two peaks: the first peak appeared at 15 min (71.69∓14.19 pmol/L) with a level significantly higher than that at 15 min following glucose load (P<0.05), and the second occurred at 90 min (55.35∓13.19 pmol/L). The area under curve of GIP showed no significant difference between the two loads (P>0.05). Compared with glucose load, mixed meal load resulted in an increase of the first GIP peak and an earlier insulin peak (30 min vs 60 min), but a significant decrease of blood glucose at 15 min (P<0.05).

CONCLUSIONCompared with glucose load, mixed meal (containing fat) can strongly stimulate GIP release and cause earlier occurrence of the insulin peak, which might be an important reason for the lower blood glucose after mixed meal.

Adult ; Blood Glucose ; metabolism ; China ; ethnology ; Diet ; Energy Intake ; Female ; Gastric Inhibitory Polypeptide ; secretion ; Glucose Tolerance Test ; Humans ; Insulin ; secretion ; Male ; Middle Aged ; Young Adult

OBJECTIVEMany studies have shown that tissue development is closely correlated with fluid transport. Aquaporins (AQPs) are a group of cell membrane proteins that actively and selectively transport water. This study aimed to investigate the changes of AQPs expression during lung development in rats in order to elucidate the role of AQPs in the rat lung development.

METHODSAQP1, AQP3, AQP4 and AQP5 proteins and mRNA in the lung cell membrane were measured by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) respectively in the 20-day-old embryo (E20), 7-day-old newborn rat, and one-month-old young and adult rats. The correlation between AQPs expression and lung development was studied.

RESULTSWith increasing age, the lung development showed a dynamic and successive course, with the most rapid from the fetus to the newborn rat, and then a slowed down afterwards. AQPs mRNA was weakly expressed in the lung of the E20 group. Lung AQPs mRNA and protein increased rapidly after birth until adulthood. The AQPs distribution patterns in the lung were unique with no duplication. There was a positive correlation between AQPs expression and lung development (P<0.05).

CONCLUSIONSIn addition to being involved in the transepithelial transport of water in the lung, AQPs is also related to its development.

Animals ; Aquaporins ; analysis ; genetics ; physiology ; Immunohistochemistry ; Lung ; embryology ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the expression change of interleukin-8 (IL-8) gene in the basilar artery of rabbit and the effect of IL-8 on the development of cerebral vasospasm induced by experimental subarachnoid hemorrhage (SAH).

METHODSThirty five healthy Japanese White Rabbits were randomly divided into saline-control group and experimental group. The experimental group was subdivided into four groups, representing day 1, 4, 7 and 14 after the first blood injection of SAH. The delayed cerebral vasospasm (DCVS) model was established by double injection of autologous blood into the cisterna magna. The expression change of cytokine IL-8 mRNA in the basilar artery was analyzed by RTPCR.

RESULTSThe expression of IL-8 gene increased on day 4-7 after the first blood injection of SAH compared with control (P< 0.001), and decreased to normal on day 14. The expression of IL-8 gene in the SAH groups were positively correlated with the degree of basilar artery stenosis (r = 0.642, P< 0.01).

CONCLUSIONThe expression level of IL-8 gene in basilar arteries was intimately associated with the degree of cerebral vasospasm, suggesting that IL-8 may play an important role in the DCVS after SAH as an immunological inflammatory factor.

Animals ; Basilar Artery ; metabolism ; Disease Models, Animal ; Gene Expression Regulation ; physiology ; Interleukin-8 ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Subarachnoid Hemorrhage ; metabolism ; pathology ; Time Factors

Objective To study the genomic genotypes and variation of human enterovirus 71(EV71)infected infants in Guangzhou city,in 2008 and 2010.Methods Primers were designed on the basis of the genomic sequence of EV71 SHZH03 strain(AY465356)in the GenBank,and EV71genome amplified by RT-PCR.PCR-products were directly sequenced and the genomic nucleotide sequences were analyzed with the programs of Clustal W/X,DNASTAR and MEGA 4.1.Results 9strains of EV71 genome appeared to be 7405 bp in length.The genomic sequences of EV71Guangzhou strains were compared with those of EV71 in GenBank,which revealed that the homology with EV71 genotype C4a Fuyang strains ranged between 98%-99%.Homology with genotype C4b were 92%-94%,with genotypes C1,C2,C3 as 82%-83%,with genotypes B3,B4,B5 as 81%-83%and the homology with genotype A was 80%.When compared the VP1 genes of EV71 Guangzhou strains with genotypes A,B,C virus,we revealed that the highest homology was also with genotype C4a.When compared the VP1 amino acid sequences of EV71 Guangzhou strains with genotype A,B,C virus by Clustal W program,the results revealed that the amino acid residue Q at position 22 in VP1gene was transformed to H,while 213(S→T)and 1764(V→(Ⅰ))mutations in polyprotein were discovered.Conclusion Data from the sequences and phylogenetics analysis on those Guangzhou strains in 2008 and 2010 revealed that those isolates belong to genotype C4a,with the homology with Fuyang strains as 98%-99%.Mutation of amino acid residue H at position 22 in VP1 gene was discovered and the neutralizing antibody of EV71 might have been conversed by this residue.213(S→T)and 1764(V→Ⅰ)mutations in polyprotein were also discovered.