3.Clonality analysis for differentiating multicentric origin and intrahepatic metastasis in multiple hepatocellular carcinomas
Jian WANG ; Yan SUN ; Hong ZHENG ; Yunlong CUI ; Qiang LI
Chinese Journal of Hepatobiliary Surgery 2010;16(12):906-909
Objective To explore the differential diagnostic significance of clone analysis for multicentric occurrence (MO) and intrahepatic metastasis (IM) in hepatocellular carcinomas (HCCs).Methods Loss of heterozygosity (LOH) and microsatellite instability (MSI) were analyzed by microsatellite polymorphism test and the integration sites of HBV were assessed by Southern blot in each nodule of the HCCs. The clonalities were then compared between each nodule of the same patient and the diagnosis of MO or IM was concluded. Finally, the results based on clonality analysis were compared with those according to clinicopathological and imaging data. Results According to the results of LOH and MSI in 79 nodules and nontumorous tissue from 35 cases of mutiple HCCs, 5 (14.3%)and 29 cases (82.9 %) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The integration sites of HBV could be analyzed in 77 nodules from 34 multiple HCCs. Among them, 6 (17. 6%) and 27 cases (79.4%) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The classification results of microsatellite polymorphism test and HBV integration sites analysis demonstrated a significant positive correlation (rs = 0.909, P<0.001). Nevertheless, neither the classification of microsatellite polymorphism test nor that of HBV integrate sites analysis was correlated with the discrimination according to clinicopathologic and imaging data (rs=0. 133, P=0. 468, rs =0. 262, P=0. 155, respectively). Recurrence in patients in the MO group occurred significantly later than that in IM cases who were diagnosed by clonality analyses (P=0. 001). Conclusion The clonality analysis based on the results of LOH and MSI or assessments of HBV integrate sites is useful for the differential diagnosis of MO and IM and their treatment and prognosis.
4.Virulence gene detection and multi-locus sequence typing of Vibrio parahaemolyticus from patients in Ningbo,China
Hong GAO ; Qifa SONG ; Jingye XU ; Jian ZHENG ; Xuanyi SHEN
Chinese Journal of Zoonoses 2015;(3):240-243
To investigated the toxin genes distribution and molecular characteristics of Vibrio parahaemolyticus from pa‐tients in Ningbo ,V .parahaemolyticus strains were collected from patients with food poisoning and diarrhea .Thermostable di‐rect hemolysin gene (tdh) and TDH‐related hemolysin gene (trh) were detected by polymerase chain reaction (PCR) .Molecu‐lar characteristics were acquired by multi‐locus sequence typing (MLST ) .Of 248 clinical strains were isolated from 2006 to 2012 .Forty‐eight strains were selected to detect virulence genes and MLST genotyping .Forty‐two isolates were detected as tdh+ and 11 isolates were detected as trh+ .There were 9 STs and one undifferentiated type in Ningbo clinical strains .Thirty‐two strains were classified into ST3 ,5 strains into ST265 and 3 strains into ST120 .ST265 was found in Ningbo strains com‐pared with strains from other regions of China .Strains with tdh+ accounted for the majority in Ningbo clinical strains .Twen‐ty‐five strains of ST3 clone were tdh+ /trh‐.There were 9 STs coexsited in Ningbo clinical strains .ST3 clone was dominant , followed by ST265 and ST120 .Strains with tdh+ /trh‐were dominant in the ST3 clone .The unique ST262 was found in Ning‐bo clinical strains .
5.Effects of selective deletion of adenosine A_(2A) receptors of peripheral leukocytes on cerebral white matter injury induced by chronic hypoperfusion
Wei DUAN ; Jian ZHENG ; Li GUI ; Hong TIAN
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective To investigate the effects of deletion of adenosine A2A receptors on peripheral leukocytes on cerebral white matter lesions induced by chronic cerebral hypoperfusion.Methods Twenty-four wild type(WT) male mice were given a ? irradiation of 12.5Gy,followed by receiving bone marrow cells tail vein from female A2A receptor knocked out(KO) mice via tail vein,were assigned as KO→WT group,while those received bone marrow cells from WT female mice were assigned as WT→WT group(n=20).The efficiency of reconstitution of bone marrow cells in recipient mice was assessed 7 weeks after transplantation by PCR and immunofluorescent technique.Then,the recipient mice were subjected to bilateral common carotid artery stenosis with internal diameter of 0.18mm by external banding using microcoils at 8 weeks after transplantation.On 7d,14d and 30d after the surgery,corpus callosum,fiber bundles of Caudoputamen and optic tract were harvested from the cerebral white matter,and stained with Kluver-Barrera staining for observing the changes in nerve fibers,and with GFAP and CD11b immunohistochemistry staining for observing the proliferation of microglia and astrocytes.Results At 7 weeks after successful transplantation,the genotype of sex chromosome in peripheral leukocytes of the male recipient mice was changed into female pattern.The expression rate of A2A receptor was 9.73%?2.05% in KO→WT group and 93.82%?11.24% in WT→WT group,with significant difference between the two groups(P
6.Behavioral and pathologic changes of chronic cerebral hypoperfusion in mice
Wei DUAN ; Jian ZHENG ; Li GUI ; Hong TIAN
Journal of Third Military Medical University 2003;0(09):-
Objective To observe behavioral characteristics and pathologic changes of chronic cerebral hypoperfusion in mice. MethodsTotally 62 adult C57BL/6 mice were subjected to either sham-operation (n=31) or bilateral common carotid artery stenosis using external microcoils with an inner diameter of 0.18 mm(n=31). At 30 d after the stenosis, the animals of the 2 groups (8 mice for each group) underwent behavioral test of 8-arm radial maze. In 7, 14 and 30 d, the rest mice were sacrificed for their brain tissue samples which were examined with Kluver-Barrera staining and immunohistochemical assay for markers of microglia and astroglia, glial fibrillary acidic protein (GFAP) and CD11b. In 14 d after the model establishment, Evans blue dye extravasation test was performed for the blood-brain barrier function. ResultsThe model group made significantly more errors than sham-operated group in 8-arm radial maze test at 30 d after the surgery. White matter lesions occurred and the proliferation of activated microglia and astroglia were observed in white matter in model mice after 14 and 30 d bilateral common carotid artery stenosis. The disruption of blood-brain barrier function of model mice was indicated in the evans blue extravasation test at 14 d after bilateral common carotid artery stenosis. ConclusionCognitive impairment, white matter lesions and glial activation are successfully induced after bilateral common carotid artery stenosis in mice model of chronic cerebral hypoperfusion.
7.Role of adenosine A_(2A) receptor in ischemic brain injury induced by transient middle cerebral artery occlusion in mice
Hong TIAN ; Wei DUAN ; Li GUI ; Jian ZHENG
Journal of Third Military Medical University 2003;0(10):-
Objective To investigate the effect of adenosine A2A receptor deficiency on the ischemic neuronal injury and its potential mechanism.Methods Transient(2 h)cerebral ischemia was induced by the middle cerebral artery occlusion(MCAO)in mice.Adenosine A2A receptor knockout(A2ARKO)mice and their wild-type littermates(A2ARWT)were divided into 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group.Cerebral infarction volume was measured by image analysis of brain sections stained with cresyl violet(CV).Brain water content was evaluated with the dry-wet weighing method.The expression of calbindin D-28k(CB)and aquaporin-4(AQP4)in ischemic brain was determined with immunohistochemical methods.Results The cerebral infarction volumes in 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group of A2ARKO mice were lesser than those in the corresponding groups of A2ARWT mice.Compared with A2ARWT mice,A2ARKO mice had more CB,lesser AQP4 expressions and lesser brain water contents.Conclusion Adenosine A2A receptor deficiency exerts the protection against ischemic brain injury both in the acute phase and reperfusion phase,and attenuates brain edema caused by cerebral ischemia,which may be due to the inhibition of intracellular calcium overload and AQP4 expression.
8.Optimization of the Extraction Technology of Oleanolic Acid from Elsholtzia Bodinieri V.by Orthogonal Test
Haobin HU ; Hong CAO ; Xudong ZHENG ; Yufeng JIAN ; Jianxin LIU
China Pharmacy 2001;0(09):-
OBJECTIVE:To optimize the extraction technology of oleanolic acid from Elsholtzia bodinieri V.METHODS:The extraction technology was optimized by orthogonal-test using microwave-assisted dynamic extraction method with oleanolic acid as index;And the content of oleanolic acid was determined by TLC-spectrophotometry.RESULTS:The opti-mum extraction condition was described as follows,the microwave power is600W with methanol as extracting solvent,the flow rate was8ml/min and the inner diameter of extraction tube was10mm.CONCLUSIONS:The microwave-assisted dynamic extraction method is simple,safe,rapid,accurate and with high extraction yield.The optimized extraction technology is stable and feasible,and suitable for industrialized production.
9.Comparison on eye biometry of Lenstar 900, A-scan ultrasound and keratometer in patients with cataract
Dan, HU ; Gang-Ping, ZHAO ; Jian-Hong, YU ; Xiao, ZHENG
International Eye Science 2014;(8):1440-1443
AIM:To investigate the differences among Lenstar 900, A-scan ultrasound and keratometer in measurement of axial length ( AL ) , anterior chamber depth ( ACD ) and corneal curvature ( K1 , K2 , Km ) , and evaluate the consistency of the instruments, with the purpose providing references for the clinical application of Lenstar 900.
METHODS: In this study we picked up 36 patients ( 50 eyes ) underwent cataract surgery, and lens nucleus hardness were under level IV. Before the operation, AL, ACD and K1 , K2 , Km were measured by Lenstar 900, A-scan ultrasound and keratometer respectively. The differences between the results were compared by the paired t-test. The correlation of the results was analyzed by Pearson correlation analysis, and the consistency was measured by Bland-Ahamn method.
RESULTS: The mean AL and ACD values measured by Lenstar 900 and A-scan ultrasound had no significantly statistic differences (P>0. 05). The K1, K2, Km measured by Lenstar 900 and keratometer were not significantly statistical different (P>0. 05). The results measured by these three instruments had close linearity correlation ( r>0.9, P<0. 01). The consistency of the results was well in Bland-Ahamn analysis.
CONCLUSION:The preoperatively biometric result of Lenstar 900, A - scan ultrasound and keratometer in patients with cataract are all reliable, and they can be substituted by each other. However, Lenstar 900 can not only measure AL, ACD and corneal curvature at the same time, but also cornal thickness, lens thickness, white to white, pupil size, optical axis eccentricity, retinal thickness and so on. It has a number of advantages such as non-touching, convenient and efficient, and can be recommended to use widely.
10.Mutation of X region nucleotide sequence of hepatitis B virus DNA in patients with HBV reinfection after Hver transplantation
Zhongyang SHEN ; Hongli SONG ; Hong ZHENG ; Jian WANG ; Weiping ZHENG ; Zhenglu WANG ; Fan ZHANG
Chinese Journal of General Surgery 2008;23(5):368-371
Objective To analyze the mutation of HBV X region nucleotide sequence in patients with HBV reinfection after liver transplantation. Methods In this study 320 patients received liver transplantation due to HBV-related end stage hver diseases between June 2002 and Dec 2003.Postoperatively polymerase chsin reaction was used to aInplify their serum HBV DNA fragments for direct sequence analysis.Patients that were followed-up for 1.5~3 years were enlisted for analysis.Results All the 11 reinfection recipients showed nucleotide mutations in X region mng4ng from 5 to 39 sites after transplantation.An A to T mutation at nt1762 and G to A mutation at nt1764 were found in 6 cases.The mutations at nucleotide fnt)1636~1741 were found in all 11 cases.Condusions The results indicated that mutations of nt 1762 and nt 1764 are very common and immunosuppressants cannot change the mutations in patients with HBV reinfection after liver transplantation.