2.Effects of 1,25(OH)_2D_3 on cell proliferation in mouse osteoblasts
Qing-Xian TIAN ; Gong-Yi HUANG ;
Chinese Journal of Geriatrics 2003;0(12):-
Objective To investigate the effects of 1,25(OH)_2D_3 on cell proliferation and cell cycle progression in mouse osteoblasts.Methods Sterile bones of skull of mouse were taken from 30 newborn mouse,and the osteoblast were separated by enzyme digestion methods.After 1,25(OH)_2D_3 in different concentrations were added into culture medium,the effects of 1,25(OH)_2D_3 on cell proliferation of mouse osteoblasts and on cell cycle progression were examined by mono-nuclear celldirect cytotoxicity assay(MTT)reduction assay and flow cytometry respectively.Results After 24,48,72 h of 1,25(OH)_2D_3 incubation,the cell number of osteoblast had significant difference among groups of 1,25(OH)_2D_3 of 10~(-8),10~(-9),10~(-11)mol/L.Significant differences were found in the cell cycle progression in response to 1,25(OH)_2D_3 treatment from the Gl(84.30?1.90)to the G2-M (7.70?0.667)and S(8.00?1.42)phases when compared with those in the control group. Conclusions Cell proliferation of mouse osteoblasts can be inhibited by 1,25(OH)_2D_3 in a concentration-dependent manner.
4.Expression of Thraustochytrium sp.FJN-10 ?~4-Desaturase Gene in Saccharomyces cerevisiae
Jin-Qing CHEN ; Xian-Zhang JIANG ; Li-Xia LIU ; Jian-Zhong HUANG ;
Microbiology 1992;0(06):-
1.6 kb ?4-desaturase gene(FAD4)was amplified by PCR using plasmid pGEM-TFAD4 as template.The fragment was subcloned into the HindⅢ/XbaⅠrestriction site of pYES2.0 vector.Recombinant plasmid pYFAD4 was transformed into Saccharomyces cerevisiae strain INVScl for expression.It was found to exhibit ?4-fatty acid desaturase activity in the recombinant S.cerevisiae YFAD4 in the presence of exogenous fatty acid substrate docosapentaenoic acid(100?mol/L)under introduction of GAL1.Expression of the FAD4 under appropriate media and temperature conditions led to the production of DHA and it reached 41.13% of the total yeast fatty acid by GC detection.It was suggested that the protein encoded by FAD4 could specifically catalyze DPA into DHA.
5.Construction and Characterization of a ppbR gene Mutant of Pseudomonas sp. M18
Fei ZHENG ; Xian-Qing HUANG ; Yu-Quan XU ; Xue-Hong ZHANG ;
Microbiology 1992;0(02):-
Pseudomonas sp. M18, one of plant-growth-promoting rhizobacteria, can produce secondary metabolites including phenazine-1-carboxylic acid (PCA) and pyoluteorin (Plt). PA2572 gene coding protein is a probable two-component response regulator in Pseudomonas according to homologous speculations. In order to investigate its genetic function, PA2572 homologous gene, ppbR, was amplified from M18 genome, inactivated by inserting a Gm cassette. The resulting reconstruct was introduced into the M18 genome using homologous recombination technique, so as to obtain the null mutant M18P. The results showed that the M18P has less flagellar swimming and swarming motility, and yielded fewer PCA. The production of PCA was only 50% of the wild type. However, there was no remarkable difference between mutant and wild type in producing pyoluteorin in KMB medium.
6.Cloning and prokaryotic expression analysis of HDS from Salvia miltiorrhiza bge.f.alba.
Dan JIANG ; Qi-Xian RONG ; Qing-Jun YUAN ; Wen-Jing ZHANG ; Yong-Qing ZHANG ; Lu-Qi HUANG
Acta Pharmaceutica Sinica 2014;49(11):1614-1620
According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular
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DNA, Complementary
;
genetics
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Diterpenes, Abietane
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biosynthesis
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Enzymes
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biosynthesis
;
genetics
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Escherichia coli
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metabolism
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Plant Proteins
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biosynthesis
;
genetics
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Salvia miltiorrhiza
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enzymology
;
genetics
7.Effects of n-hexane exposure on human serum myelin basic proteins.
Wei ZHOU ; Juan YI ; Hui-Ping HUANG ; Ying-Ping XIANG ; Jia-Xi HE ; Qing-Jun LIU ; Xian-Qing HUANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(6):421-423
OBJECTIVETo explore the effects of n-hexane on expression of serum myelin proteins (MBP) in workers occupationally exposed to n-hexane.
METHODSIn this study, 269 workers exposed to n-hexane for more than one year and 104 subjects not exposed to n-hexane served as the exposure group and the control group, respectively. The urinary 2,5-hexanedione levels in all subjects were detected. On the basis of urinary 2,5-hexanedione levels, the exposure group was divided into the high exposure sub-group and low exposure sub-group. The serum myelin basic protein (MBP) levels were measured by ELISA kit.
RESULTSThe mean concentration of urinary 2,5-hexanedione in the exposed group was (3.10 +/- 1.35) mg/L. The concentration of urinary 2,5-hexanedione in the control group was undetectable. The levels of serum MBP in the high exposure sub-group and low exposure sub-group were (2.43 +/- 0.24) and (1.62 +/- 0.23) microg/L, respectively, which were significantly higher than that (0.78 +/- 0.12) microg/L in the controls (P < 0.01). Pearson correlation analysis showed the positive correlation between serum MBP levels and urinary 2,5-hexanedione levels (r = 0.781, P < 0.01).
CONCLUSIONThe results of present study showed that the serum MBP levels of workers occupationally exposed to n-hexane significantly elevated, and the serum MBP can serve as the effective biomarker of n-hexane exposure.
Adolescent ; Adult ; Case-Control Studies ; Female ; Hexanes ; adverse effects ; Humans ; Male ; Myelin Basic Protein ; blood ; Occupational Exposure ; Young Adult
8.Security evaluation of bupivacaine, ropivacaine combined with fentanyl in postoperative continuous epidural analgesia.
Xiao-hong WAN ; Qing-qing HUANG ; Mei-xian SU ; Lin-jun WAN ; Hai-qiong HUANG
Chinese Journal of Surgery 2006;44(17):1200-1202
OBJECTIVETo investigate the effects, side-effects and security of bupivacaine, ropivacaine combined with fentanyl in postoperative continuous epidural analgesia.
METHODSA total of 1 600 postoperative continuous epidural analgesia patients receiving different agents in SICU were divided into two groups: 0.1% bupivacaine +5 microg/ml fentanyl group (group B, n = 920) and 0.2% ropivacaine +2 microg/ml fentanyl group (group R, n = 680). The effects (visual analog-scale score and content to analgesia), side effects were analyzed retrospectively in the two groups.
RESULTSCompared with group B, patients in group R had higher analgesia contentment (P < 0.05), but no difference in visual analog-scale score was found in the two groups. The incidences of urinary retention, nausea and vomiting, skin itching in group B were significantly higher than those in group R (P < 0.05). In each group, patients over sixty had higher ratio of hypotension than those under sixty (P < 0.05); The female patients had a higher incidence of nausea and vomiting than male patients (P < 0.05); The incidence of debility and numbness of lower limbs in patients with lumbar segments epidural analgesia was higher than those with thoracic analgesia (P < 0.05).
CONCLUSIONS0.1% bupivacaine +5 microg/ml fentanyl and 0.2% ropivacaine +2 microg/ml fentanyl can provide adequate pain relief in postoperative continuous epidural analgesia, and 0.2% ropivacaine +2 microg/ml fentanyl comes with less side effects. The incidence of complication is related with analgesics, age, gender and the position of epidural puncture.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Amides ; administration & dosage ; adverse effects ; Analgesia, Epidural ; adverse effects ; methods ; Analgesia, Obstetrical ; adverse effects ; methods ; Analgesics, Opioid ; administration & dosage ; adverse effects ; Anesthetics, Local ; administration & dosage ; adverse effects ; Bupivacaine ; administration & dosage ; adverse effects ; Female ; Fentanyl ; administration & dosage ; adverse effects ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; prevention & control ; Retrospective Studies
9.Application of radiotherapy-assistance fixation device in salivary gland scintigraphy
Zhi-Yong SUN ; Jing XIAN ; Qing-Xu WANG ; Xian MENG ; Ye HUANG ; Xia WU ; Jian-Zhong CHEN
Chinese Medical Equipment Journal 2018;39(4):67-70
Objective To apply a simple fixator to salivary gland scintigraphy to evaluate its effect on body position movement. Methods Totally 30 female patients complaining xerostomia were randomly and equally divided into a conventional scan group and a simple fixator group,who were injected with Technetium Tc-99m Pertechnetate intravenously. In the conventional scan group the patients had their necks fixed with the bracket and cushion,while in the other group the fixation was executed with the single fixator. SPECT imager was used for dynamic salivary gland scintigraphy, and two nuclear medicine physicians evaluated the maximum shift of the parotid gland along left-right(X)and head-foot(Y)axes in series of dynamic images. SPSS 16.0 software was used for data analysis. Results In the conventional scan group the maximum shift was(6.6±4.6)mm at X axis and(5.2±3.4)mm at Y axis;in the single fixator group the maximum shift was(3.2± 1.6)mm at X axis and(3.0±1.3)mm at Y axis.There were significant differences between the maximum shifts in the two groups (P=0.012,X axis;P=0.027,Y axis).Conclusion The single fixator assists in salivary gland scintigraphy,and alleviates the body position movement during dynamic acquisition and provides data support for functional parameter calculation and result determination.