1.Effects of different doses of topiramate on expression of NCAM and GAP- 43 mRNA in hippocampus of rats with epilepsy
Danhong WU ; Ping ZHONG ; Guanshui BAO ; Hong ZHOU ; Jun SHEN ; Feifei HUANG
Journal of Shanghai Jiaotong University(Medical Science) 2009;29(12):1459-1462
Objective To explore the effects of different doses of topiramate (TPM) on the expression of nerve cell adhesion molecule (NCAM) and growth-associated protein 43 (GAP- 43) mRNA in hippocampus of rats with epilepsy. MethodsForty-eight rats were randomly divided into normal control group, kainic acid (KA) group, 10 mg/kg TPM group, 40 mg/kg TPM group, 100 mg/kg TPM group and 400 mg/kg TPM group (n=8). The models of rats with epilepsy treated by different doses of TPM were established. The behavior of rats was observed, and the expression of NCAM and GAP- 43 mRNA in hippocampus of rats was determined by Real-time PCR. Results The expression of NCAM and GAP- 43 mRNA in KA group was significantly higher than that in normal control group (P<0.01), while there was no significant difference between 10 and 40 mg/kg TPM groups and KA group, that in 100 and 400 mg/kg TPM groups was significantly lower than that in KA group (P<0.01), and that in 400 mg/kg TPM group was significantly lower than that in 100 mg/kg TPM group (P<0.01). Conclusion KA can up-regulate the expression of NCAM and GAP- 43 mRNA in hippocampus of rats with epilepsy. Higher dose of TPM can inhibit the expression of NCAM and GAP- 43 mRNA, and the inhibitory effect is related with the dose of TPM.
2.Eukaryotic expression and DNA immunization of hemolysin HlyX gene from Leptospira serovar Lai
Bi HUANG ; Lang BAO ; Qi ZHONG ; Zhengling SHANG ; Huidong ZHANG ; Ying ZHANG
Chinese Journal of Microbiology and Immunology 2008;28(2):134-138
Objective To study on the expression of the eukaryotic recombinant vector carrying HlyX gene of Leptospira serovar Lai in mammalian cell and explore the humoral immune response in BALB/c mice immunized with the recombinant plasmid. Methods The HlyX gene was amplified from Leptospira serovar Lai genomic DNA by PCR and inserted into pcDNA3.1 vector. After transformed into E. coli DH5α,the recombinant plasmid was assayed for identification by PCR analysis,restriction nuclease enzyme digestion and sequencing. The recombinant plasmid was transfected into COS-7 cells,then RT-PCR and Western blot were performed to test the expression of the target gene. The recombinant plasmid was injected intramuscularly into BALB/c mice for three times at intervals of two weeks,and the antibody titer was measured by ELISA. Results PCR showed the full length HlyX gene was about 1100 bp. PCR analysis,restriction nuclease enzyme digestion and sequencing indicated the recombinant vector was constructed successfully. After the plasmid Was transfected into COS-7 cells,a fragment about 1100 bp was found by RT-PCR and a specific band relative molecular mass(Mr)about 40×103,which was consistent with the expected size of the target proteins was showed by Western blot. ELISA showed the antibody titer in BALB/c mice immunized by the HlyX gene of Leptospira serovar Lai can elicit high-titer antibody in BALB/c mice,which has laid the foundation for the application of the DNA vaccine.
3.Overexpression of Alcohol DehydrogenaseⅠ in Saccharomyces cerevisiae
Li-Na QIN ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.
4.Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium
Zhi-Ping LI ; Xian-Zhang JIANG ; Bao-Yu TIAN ; Zheng-Yu SHU ; Jian-Zhong HUANG ;
Microbiology 1992;0(02):-
Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.
5.Short-term results of cutting balloon and intravascular brachytherapy for the treatment of coronary in-stent restenosis
Fei YE ; Shao-Liang CHEN ; Bao-Xiang DUAN ; Jin HUANG ; Zhi-Zhong LIU ; Jie SONG ;
Chinese Journal of Clinical Pharmacology and Therapeutics 1999;0(04):-
0.05); the levels of LL, LI, RRS in CBA group and CBA+IBT group were significantly lower than those in control group(P
6.The patterns of head and neck cancer incidence in Shanghai from 2003 to 2012
Chunxiao WU ; Pingping BAO ; Zhezhou HUANG ; Minlu ZHANG ; Kai GU ; Yongmei XIANG ; Peng PENG ; Yangming GONG ; Ying ZHENG ; Weijian ZHONG
China Oncology 2017;27(6):406-414
Background and purpose:Head and neck cancer is common worldwide. This article aimed to describe the patterns of incidence of head and neck cancer and their changes in Shanghai from 2003 to 2012, in order to provide reference for prevention programs, research and control strategies on cancer.Methods:The data of lip, oral cavity and pharynx cancer cases were collected by the Shanghai Cancer Registry. The distributions of Shanghai lip, oral cavity and pharynx cancer incidences from 2003 to 2012 were described. The patterns were described according to gender, age, basis of diagnosis, histological type, diagnostic stage in detail. We compared incidences of recent 5 years (from 2008 to 2012) with those of previous 5 years (from 2003 to 2007).Results:On average from 2003 to 2012, 1105 new cases of head and neck cancer were diagnosed in Shanghai each year, with 2.08% among the total cancer cases. The crude rate was 8.01 per 100000 and the age-standardized rate was 4.45 per 100000. Nasopharyngeal cancer was the major subtype of the head and neck cancer, with 50.81% among the total head and neck cases. The crude and age-standardized rates among males were higher than those among females. The histologically verified percentage was 85.77%. The squamous carcinoma was the major histological type, with 57.58% among the total cases. The age-stan-dardized rate of nasopharyngeal cancer was in decline.Conclusion:The incidence of head and neck cancer was low in Shanghai during the past 10 years. Male cases were more than female cases. The major histological type was squamous carcinoma. Half of new cases were nasopharyngeal cancer which appeared to affect patients at a relatively young age. Patients with nasopharyngeal cancer were diagnosed at relatively advanced stages.
7.Diagnostic features of pathological fractures located in extremities caused by bone tumor or tumor like lesion
Yan-cheng LIU ; Yong-cheng HU ; Qun XIA ; Bao-shan XU ; Ning Jl ; Hong-chao HUANG ; Zhong-li ZHANG
Chinese Journal of Orthopaedics 2011;31(9):944-948
ObjectiveTo explore the clinical characteristics of pathological fracture in extremities caused by bone tumors or tumor-like lesions. MethodsFrom August 2002 to December 2010, 139 patients with pathological fractures were entered in the study, including 79 males and 60 females with an average age of 31.1 years. Fractures included tumor-like lesion in 55 cases, benign tumor in 13, giant cell tumor (GCT)in 7, primary malignant tumors in 28, and metastatic tumors in 36. Forces induced to fractures were classified into four grades: spontaneous fracture, functional activity, minor injury, severe injury. Age, fracture location, histological results, fractures forces, prodromes, and misdiagnosis were all observed. Chi-square test were use to compare forces and prodromes within different tumors. ResultsThe highest morbidity rate is 32.4%(45/139) which lies in 11-20 years old. The cites of fractures including femurs in 71 cases, humerus in 36, tibia in 15, fingers in 7, radiuses in 4, fibula in 3, ulnas in 2, and metatarsus in 1. Fracture forces include spontaneous fractures in 29 cases, functional activity in 42, minor injuries in 65, and traumatic injuries in 3. Sixty-seven patients(48.2%) had local prodromes. The prodromes of both malignant tumors and metastatic tumors were more than benign tumors. Twenty cases experienced misdiagnosis with average delay time of 12 weeks. ConclusionMinor injury forces and local prodromes are clinical key features of pathological fractures. Both of them are key points of avoiding misdiagnosis.
8.Application of improved submental island flap in hypopharyngeal cancer reserved laryngeal function surgery.
Pingqing TAN ; Jie CHEN ; Wenxiao HUANG ; Ronghua BAO ; Jinyun LI ; Junqi WANG ; Li XIE ; Waisheng ZHONG ; Hailin ZHANG
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2015;29(15):1342-1345
OBJECTIVE:
This study aimd to evaluate the application and clinical effect of improved submental island flap in hypopharyngeal cancer reserved laryngeal function surgery.
METHOD:
A retrospective review of clinical data was performed on 38 patients of hypopharyngeal cancer reserved laryngeal function using sumental island flaps, by the way of improving in design of vascular pedicle, reconstructive mode of laryngeal and hypopharyngeal function and closing of wound of neck following hypopharyngeal cancer resection. Meanwhile, the effect and prognosis was comprehensively assessed on patients with hypopharyngeal cancer reserved laryngeal function using improved submental island flaps.
RESULT:
The submental flaps kept alive in all 38 cases. During the follow-up period, 18 cases were dead, and of them, 7 cases died of the second primary carcinoma, included 4 cases of esophagus cancer, 1 case of cancer of soft palate, 2 cases of nasopharyngeal carcinoma; and 5 cases died of cervical or parapharyngeal lymph nodes recurrence; 2 cases died of hepatic metastasis; and 4 cases died of pulmonary metastasis. The overall 5-years survival rate was 52.6%.
CONCLUSION
Improved submental island flap repairing postoperative defect of hypopharyngeal cancer reserved laryngeal function has many advantages including higher success rate, more security, easy and simple to operate as well as good clinical effects, and is worth to widespread using.
Humans
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Hypopharyngeal Neoplasms
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surgery
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Hypopharynx
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surgery
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Larynx
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Neck
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surgery
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Neoplasm Recurrence, Local
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Prognosis
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Reconstructive Surgical Procedures
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Retrospective Studies
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Surgical Flaps
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Survival Rate
9.Characteristics of fruit ripening in tomato mutant epi.
Zhong-feng WANG ; Tie-jin YING ; Bi-li BAO ; Xiao-dan HUANG
Journal of Zhejiang University. Science. B 2005;6(6):502-507
The characteristics of fruit ripening and expression of ripening-related genes were investigated in epi, an ethylene overproduction mutant of tomato (Lycopersicon esculentum Mill.). The epi produces apparently more ethylene than its wild type VFN8 at every stage of vegetative and fruit growth and ripening; compared to VFN8, the epi fruit showed higher CO2 evolution, faster descending of chlorophyll, slightly quicker increase of carotenoid and lycopene, and faster reduction in pericarp firmness during maturation and ripening; and the mRNAs of three ripening-related genes including E8, pTOM5 and pTOM6 were at higher levels in epi. The ripening-related characteristics changing of the fruit are consistent with the increase of ethylene production and ripening-related genes expression. These results suggest that epi mutation possibly did not affect the ethylene perception and signaling during fruit ripening, and that the modified characteristics of fruit ripening possibly resulted from the ethylene overproduction and increased expression of ripening-related genes.
Ethylenes
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metabolism
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Fruit
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growth & development
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metabolism
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Gene Expression Regulation, Plant
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physiology
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Hardness
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Lycopersicon esculentum
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growth & development
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metabolism
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Mutation
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Plant Proteins
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metabolism
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Signal Transduction
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physiology
10.Aggressiveness and extent of prostatic inflammation relates with serum PSA levels in type IV prostatitis.
Li-Bo MAN ; Gui-Zhong LI ; Guang-Lin HUANG ; Jian-Wei WANG ; Bao-Yue LIU
National Journal of Andrology 2012;18(8):710-714
OBJECTIVETo investigate the relationship of the histopathologic grade and extent of prostatic inflammation with the level of serum PSA in patients with type IV prostatitis.
METHODSWe performed transrectal ultrasound-guided prostate biopsy for 120 patients suspected of prostate cancer and included in this study only those with benign prostate hyperplasia (BPH) and prostatitis (n = 46), excluding the cases with prostate cancer and those with BPH but no prostatitis. We evaluated the relationship between prostatic inflammation and serum PSA levels based on the three-grade pathohistologic criteria for the extent, location and aggressiveness of prostatic inflammation. The serum tPSA levels, fPSA levels, % fPSA, and PSAD were compared among different groups.
RESULTSAs for the extent of inflammation, 35 of the 46 included cases were grade I (tPSA: [8.46 +/- 4.09] microg/L; fPSA: [1.75 +/- 0.93] microg/L; PSAD: 0.15 +/- 0.11), 7 were grade II (tPSA: [15.26 +/- 5.26] microg/L; fPSA: [2.54 +/- 0.72] microg/L; PSAD: 0.26 +/- 0.07) and 4 were grade III (tPSA: [21.05 +/- 7.58] microg/L; fPSA: [3. 19 +/- 1.13] microg/L; PSAD: 0.42 +/- 0.19), with statistically significant differences among the three groups in the levels of tPSA (P = 0.001), fPSA (P = 0.008) and PSAD (P < 0.001). Regarding the location of inflammation, 19 cases were grade I, 17 were grade II and 10 were grade II, with no significant differences in tPSA, fPSA and %fPSA among the three grades (P > 0.05). As for the aggressiveness of inflammation, 32 cases were grade I (tPSA: [8.37 +/- 4.07] microg/L; fPSA: [1.76 +/- 0.93] microg/L; PSAD: 0.14 +/- 0.11), 10 were grade II (tPSA: [13.30 +/- 5.69] microg/L; fPSA: [3.27 +/- 2.21] microg/L ; PSAD: 0.25 +/- 0.06) and 4 were grade III (tPSA: [21.05 +/- 7.58] microg/L; fPSA: [3.19 +/- 1.13] microg/L; PSAD: 0.42 +/- 0.19), with statistically significant differences among the three grades in the levels of tPSA (P = 0.002), fPSA (P = 0.024) and PSAD (P < 0.001). The extent of inflammation was positively correlated with the levels of tPSA (r = 0.6, P < 0.001), fPSA (r = 0.5, P = 0.001) and PSAD (r = 0.6, P < 0.001), and so was the aggressiveness of inflammation (tPSA: r = 0.5, P < 0.001; fPSA: r = 0.4, P = 0.008; PSAD: r = 0.7, P < 0.001), but a negative correlation was found between the aggressiveness of inflammation and %fPSA (r = -0.4, P = 0.013).
CONCLUSIONThe aggressiveness and extent of prostatic inflammation in asymptomatic prostatitis patients are significantly correlated with the level of serum PSA, which may help pathologists to avoid unnecessary repeated biopsies for patients with high-grade prostatitis.
Aged ; Biopsy ; Humans ; Inflammation ; Male ; Prostate ; pathology ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; pathology ; Prostatitis ; blood ; pathology ; Serum