To construct recombinant reporter plasmids containing different Alpha gene segments and Alpha1-TFEB fusion gene and to evaluate the promoter activity of the Alpha gene. Methods:Promoter regions of the Alpha gene were predicted using a software Primer 0.5. Five Alpha gene segments with different lengths and a normal TFEB gene promoter (pTFEB) were amplified via polymerase chain reaction, and recombinant reporter plasmids containing different Alpha gene segments and a normal TFEB gene pro-moter were constructed. Liposome transfection was used to transfect these vectors into the human embryo kidney 293T cells. The pro-moter activity of the Alpha gene was evaluated via luciferase assay. Meanwhile, the recombinant Alpha1-TFEB plasmid was construct-ed and transfected into the 293T cells. The TFEB expression of the recombinant Alpha1-TFEB plasmid was then detected via Western blot. Results: Recombinant reporter plasmids containing different Alpha gene segments and pTFEB were constructed successfully. Compared with the luciferase activity of pGL3-Basic, that of the groups with Alpha1, Alpha2, Alpha3, Alpha4 and Alpha5 significantly increased (P<0.01). The luciferase activity also increased significantly in the groups with Alpha1, Alpha2 and Alpha5 compared with that of the pTFEB group (P<0.01). The TFEB expression of the pGL3-Enhancer-Alpha1-TFEB was significantly higher compared with that of the pGL3-Enhancer group. Conclusion:In t(6;11) translocation RCC, the Alpha gene has a strong promoter activity and it en-hances TFEB expression. The strongest promoter activity region is in Alpha5 with a sequence from 643 bp to 693 bp.

Objective To investigate the relation between body mass index(BMI),skinfolds(SF)and relative weight(RW),arm girth(AG) and ag e of the children in order to look for the most suitable index used to diagnose children obesity in clinical work.Methods In 1496 healthy children (ranging from 1 to 13 years),their weight, height,AG, triceps skinfold(TSF), sub scapular skinfold(SSF) and abdominal skinfold(ASF) were measured and the relat ion of BMI,SF,RW,AG and age was analyzed.Results 1.Between BMI and the sum of three skinfolds(SF3) ,RW,AG,correlation coefficient was more t han 0.8(P＜0.001).2. With the increase of age, AG and ASF increase wh ile TSF and BMI decrease. 3.Skinfolds in girls of 3～6,12 years of age are thicker t han those in boys;BMI in boys of 8,9 years of age are higher than those in girls .Conc lusions 1.95 percentile of BMI and SF3 as index to diagnose children obes ity is better than single application of RW or one-site skinfold. 2. With the i ncrease of age, AG and abdominal fat increase, but triceps and total fat of body relatively decrease. 3. Fats in girls develop faster than in boys.

Objective To evaluate if the assay of platelet-associated immunoglobulin G (PAIgG) can improve the sensitivity of monoclonal antibody immobilization of platelet antigens assay (MAIPA) in immune thrombocytopenic purpura (ITP).Methods Anti-GP Ⅱb/Ⅲa and anti-GP Ⅰb/Ⅸ autoantibodies were detected by a modified MAIPA;PAIgG was detected by competitive ELISA.MAIPA and PAIgG were done simultaneously in 190 patients with thrombocytopenia,in which 132 were diagnosed as ITP according to the diagnostic criteria Kappa test was used to analyze the concordance between MAIPA and PAIgG.Results The sensitivities of PAIgG alone or in parallel with MAIPA to ITP were 32.6% and 37.2% higher than MAIPA alone respectively,while their specifieities were 51.7% and 56.9% lower respectively.The sensitivity,specificity,positive predictive value,negative predictive value of PAIgG in series with MAIPA were all not superior to MAIPA alone.The kappa values between MAIPA and PAIgG in ITP and non-immune thrombocytopenia were 0.129 and 0.012 respectively,which meant the concordance between them was poor.Conclusion The detection of PAIgG should not be used as screening before MAIPA in distinguishing immune from non-immune thrombocytopenia.

Objective: To study the alkaloid constituents of Huperzia serrata (Thunb.) Trev.. Methods: Various chromatographies were used for separation and purification of the alkaloids and spectroscopic analysis was used for determination of the chemical structure. Results: An alkaloid constituent(alkaloid A) was isolated from H. serrata . Conclusion: Alkaloid A was a new compound, named huperzinine B.

Objective To examine the effect of epigallocatechin gallate (EGCG) at different concentration (0,10,25, 50 and 100μmol/L ) on proliferation rate and apoptosis of nasopharyngeal carcinoma cell line C666-1 in vitro, and elucidate the role of silent information regulator 1 (SIRT1). Methods The proliferation rate in vitro of C666-1 cells stimulated by EGCG at increasing concentrations (0,10,25,50,and 100 μmol/L)for 24 h or at concentration of 50μmol/L for 0,6,12 h and 24 h were detected by cell counting kit-8 (CCK-8) assays. Cell were treated with EGCG at concentration of 0,25, 50 and 100 μmol/L for 24 h, cell apoptosis was anylysed by TUNEL assay and expression levels of SIRT1 protein was detected by western blotting. Results EGCG suppressed cell proliferation of C666-1 cell line in a concentration-dependent manner at concentration of 0 ,10,25,50,and 100μmol/L, and in a time-dependently when treated with 50 μmol/L for 12 to 24 h(P<0.05). After treated for 24 h with different concentration of EGCG at 0、25、50、100 μmol/L, cell apoptosis increased at concentration of 50 to 100μmol/L and expression of SIRT1 decreased in a concentration-dependently (P<0.05). Conclusion EGCG induced cell apoptosis of C666-1 cells by down-regulating SIRT1 expression.

Animals ; Astrocytes ; immunology ; metabolism ; Male ; Microglia ; immunology ; metabolism ; Neurons ; metabolism ; Oxidopamine ; metabolism ; Parkinson Disease ; immunology ; Rats ; Rats, Sprague-Dawley

AIMTo study the alkaloid constituents of Huperzia serrata (Thunb.) Trev..

METHODSChromatographic methods were used for the isolation and purification. Structure was elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSAn alkaloid constituent was isolated from H. serrata (Thumb.) Trev..

CONCLUSIONThe compound was found to be a novel phlegmariurine type alkaloid, named 8 beta-hydroxy phlegmariurine B.

Heterocyclic Compounds, 3-Ring ; chemistry ; isolation & purification ; Huperzia ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry

OBJECTIVETo study the effect of Shenfu Injection (SI) on the post-operative cognitive dysfunction in senile patients receiving the orthopedic surgeries after general anesthesia.

METHODS100 patients, 60-89 years old, waiting for selective surgery at the orthopedics department, were randomly assigned to two groups, the SI group (50 cases) and the control group (50 cases), who met the standard of the American Society of Anesthesiologists (ASA) II-III. After surgical incision, patients in the SI group were transfused with SI 200 mL (50 mL SI was added in 200 mL normal saline), while those in the control group were transfused with 200 mL normal saline at the same time. The mean arterial pressure (MAP), heart rate (HR), and oxygen saturation (SpO2) were compared between the two groups before anesthesia, after intubation, 30 min and 60 min after anesthesia, 5 min and 30 min after extubation. The time for surgery, the time of intra-operative hypotension, the volume of hemorrhage, the time for post-operative consciousness recovery, the liver function, the kidney function, and the fasting blood glucose of patients before surgery, 3 days and 7 days after operation were compared. The cognitive functions of patients were tested before surgery, 3 days and 7 days after operation. The incidence of post-operative cognitive dysfunction of patients was compared between the two groups.

RESULTSThere was insignificant difference in MAP, HR, and SpO2 between the two groups at each time point (P > 0.05). There was insignificant difference in the pre-operative cognitive function score, the time for surgery, the time of intra-operative hypotension, the volume of hemorrhage, the liver function, the kidney function, and the fasting blood glucose between the two groups ( P > 0.05). The time for post-operative consciousness recovery was obviously shorter in the SI group than in the control group, showing significant difference (P < 0.05). The post-operative 3-day and 7-day cognitive function scores were obviously better in the SI group than in the control group, showing significant difference (P < 0.05). The incidence of post-operative cognitive dysfunction of the SI group [3 days: 12% (6 cases); 7 days: 8% (4 cases) ] was significantly lower than that of the control group [3 days: 20% (10 cases); 7 days: 14% (7 cases) ] ( P < 0.05).

CONCLUSIONSI could improve the post-operative cognitive function of senile patients receiving the orthopedic surgeries after general anesthesia, lower the incidence of post-operative cognitive dysfunction rate, and shorten the time for post-operative consciousness recovery.

Aged ; Aged, 80 and over ; Anesthesia, General ; adverse effects ; Cognition ; Cognition Disorders ; etiology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Injections ; Middle Aged ; Orthopedic Procedures ; Postoperative Period

AIMTo study chemical constituents of Knoxia valerianoides Thorel et Pitard.

METHODSChromatographic methods were used for the isolation and purification. Structures was elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSTwo anthraquinone were isolated from Knoxia valerianoides Thorel et Pitard and identified as 1,3,5-trihydroxy-2-ethoxymethyl-6-methoxyl-anthraquinone (I) and 1,3-dihydroxy-2-ethoxymethyl-anthraquinone (II).

CONCLUSIONThe compound I was found to be a novel anthraquinone constituent and II was isolated from Knoxia valerianoides Thorel et Pitard for the first time.

Anthraquinones ; chemistry ; isolation & purification ; Molecular Structure ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Rubiaceae ; chemistry

AIMTo study the chemical constituents of Knoxia valerianoides Thorel et Pitard.

METHODSChromatographic methods were used for the isolation and purification. Structures were elucidated on the basis of chemical analysis and spectroscopic data.

RESULTSThree anthraguinones were isolated from K. valerianoides and identified as 1, 3, 5-trihydroxy-2-methyl-6-methoxyl-anthraguinone (kaoxiadin, I), 1, 3, 6-trihydroxy-5-ethoxylmethyl-anthraguinone (II) and 1, 3-dihydroxy-2-methylanthraguinone (rubiadin, III).

CONCLUSIONCompound II is a new anthraguinone constituent.

Anthraquinones ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Tubers ; chemistry ; Plants, Medicinal ; chemistry ; Rubiaceae ; chemistry