1.Pretreatment with docosahexaenoic acid enhances the protective effect of angiopoietin-1 on rat brain microvascular endothelial cells during oxygen glucose deprivation
Xiaobo CHEN ; Zhongyuan XIA ; Fan LEI ; Rui XUE
Chinese Journal of Clinical Nutrition 2015;23(2):103-109
Objective To evaluate the role of pretreatment with docosahexaenoic acid (DHA) in the protective effect of angiopoietin-1 (Ang-1) on rat brain microvascular endothelial cells (BMVECs) during oxygen glucose deprivation (OGD).Methods BMVECs were sub-cultured in vitro and divided with a random number table into 7 groups:normal control group,normal control + Ang-1 group,OGD group,OGD + Ang-1 group,OGD + DHA group,OGD + DHA + Ang-1 group,and OGD + DHA + GW9662 + Ang-1 group.The normal control and normal control + Ang-1 groups were cultured in DMEM containing serum,5 mmol/L glucose,and 1.25 mmol/L pyruvate;OGD groups were cultured in glucose-and serum-free DMEM.DHA (40 μmol/L) was added to OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups,and 5 μmol/LGW9662 [inhibitor of peroxisome proliferator-activated receptor gamma (PPAR-γ)] to OGD + DHA + GW9662 +Ang-1 group,before pretreatment for 1 hour in 5% CO2 and 95% air.After the pretreatment,Ang-1 (250 ng/ml)was added to normal control + Ang-1,OGD + Ang-1,OGD + DHA + Ang-1,and OGD + DHA + GW9662 +Ang-1 groups.The cells were cultured in 94% N2∶ 5% CO2∶ 1% O2 for 24 hours,except for normal control and normal control + Ang-1 groups,which were cultured in 5% CO2 and 95% air instead.The cell apoptosis rate was detected with flow cytometry,expressions of Bax,Bcl-2,caspase-3,Tie-1,Tie-2,pTie-2,pAkt,ZO-1proteins with Western blot.Results Compared with normal control group,the cell apoptosis rate in OGD,OGD + Ang-1,OGD + DHA,OGD + DHA + Ang-1,and OGD + DHA + GW9662 + Ang-1 groups were significantly increased (P =0.000,0.000,0.000,0.004,0.000);the expression levels of Bax,caspase-3,and Tie-1 were significantly enhanced (Bax:0.62 ± 0.03,0.38 ± 0.03,0.45 ± 0.03,0.26 ± 0.02,0.33 ± 0.02vs.0.16 ±0.01;caspase-3:0.76 ±0.05,0.42 ±0.04,0.52 ±0.02,0.32 ±0.02,0.40 ±0.02 vs.0.15 ±0.01;Tie-1:0.51 ±0.03,0.25 ±0.01,0.33 ±0.02,0.16±0.01,0.22±0.02 vs.0.12 ±0.01;all P=0.000);the expression levels of Bcl-2,Bcl-2/Bax,Tie-2,and pTie-2 were significantly decreased [Bcl-2:0.09±0.01,0.20±0.01,0.16±0.02,0.31±0.01,0.22±0.01 vs.0.34±0.01;Bcl-2/Bax:(14.93±1.86)%,(68.03±5.56)%,(36.93 ±2.22)%,(119.1 ±13.3)%,(64.23 ±6.07)% vs.(208.33 ±7.37)%;Tie-2:0.07±0.01,0.16±0.02,0.11 ±0.01,0.21±0.01,0.18±0.01 vs.0.26±0.01;pTie-2:0.05±0.01,0.15 ±0.01,0.07 ±0.01,0.22 ±0.02,0.16±0.01 vs.0.27 ±0.01;allP=0.000],in addition,pAkt and ZO-1 in OGD,OGD + Ang-1,OGD + DHA,and OGD + DHA + GW9662 +Ang-1 groups were also significantly reduced (pAkt:0.13 ±0.01,0.26 ±0.01,0.14 ±0.01,0.28 ±0.02vs.0.39±0.02;ZO-1:0.08±0.01,0.18±0.01,0.10±0.01,0.19±0.01vs.0.23±0.02;allP=0.000).Compared with the OGD group,OGD + Ang-1 and OGD + DHA + Ang-1 groups demonstrated significantly reduced cell apoptosis (both P =0.000).Compared with normal control + Ang-1 group,the expression levels of pTie-2,pAkt,and ZO-1 were significantly decreased in OGD + Ang-1 and OGD + DHA +Ang-1 groups (pTie-2:0.15 ±0.01,0.22 ±0.02 vs.0.52 ±0.02;pAkt:0.26 ±0.01,0.37 ±0.04 vs.0.67 ± 0.05;ZO-1:0.18 ± 0.01,0.24 ± 0.02 vs.0.39 ± 0.05;all P =0.000).Compared with OGD +Ang-1 group,OGD + DHA + Ang-1 group had significantly decreased cell apoptosis and expression levels of Bax,caspase-3 and Tie-1,and significantly increased expressions of Bcl-2,Bcl-2/Bax,Tie-2,pTie-2,pAkt,and ZO-1 (all P =0.000),while OGD + DHA + GW9662 + Ang-1 group showed no significant differences in these indexes (P =0.202,0.770,0.382,0.448,0.233,0.736,0.143,0.526,0.495,0.670).Conclusion Pretreatment with DHA may enhance the protective effect of Ang-1 on rat BMVECs under the condition of OGD,possibly via activating PPAR-γ,suppressing Tie-1 expression,and hence amplifying the activation of Tie-2.
2.A study on correlation between risk factors of cerebral vascular disease(CVD) and cognitive deficit associated with cerebral infarction
Xiaoyun XU ; Lei HUANG ; Hui HU ; Xiaobo GU
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(07):-
Objective To study the correlation between risk factors of CVD and cognitive deficit associated with cerebral infarction. Methods Cognitive function was tested in 101 cases of cerebral infarction using HDS-R at the end of 3 weeks and 6 months after onset of the stroke. Meanwhile, the data of risk factors and relative information of cerebral vascular disease were collected by using the scale of complications and inquiring the past history. The risk factors of CVD, such as neurological deficit, diabetes, hypertension, obesity, fever, heart diseases, smoking, overdrinking were compared between the cognitive deficit group and non-cognitive deficit group. Logistic regression analysis was adopted for data analysis. Results Of the 101 cases , 65 had cognitive deficit at the end of 3 weeks and 53 at the end of 6 months after onset of stroke. Age and score of scale of complications between cognitive deficit group and non cognitive deficit group were significantly different(P
3.The usage of HC visual laryngoscope for emergency tracheal intubation by unskilled junior emergency resident doctors
Longfei PAN ; Lei YU ; Honghong PEI ; Yuan FAN ; Xiaobo WANG ; Lijuan SU ; Wan HUANG
Chinese Journal of Emergency Medicine 2016;25(7):910-914
Objective To investigate the usefulness of HC visual laryngoscope designed for unskilled junior emergency resident doctors in respect of increasing the efficiency and success rate of emergency tracheal intubation.Methods A total of 80 patients needed the emergency tracheal intubation were at first divided into two groups randomly (random number).Patients in group A were operated by junior emergency resident doctors and patients in group B were operated by well experienced attending doctors.Then,the two groups of patients above were divided into two groups randomly (random number).Patients in group N were operated with conventional laryngoscope and patients in group HC were managed with HC visual laryngoscope.Then comparisons of the success rate of glottis exposure,the one-time success rate of tracheal intubation,the success rate of tracheal intubation,the complications,the number of attempt in successful intubation cases and the operative time consumed for successful intubation cases among the groups were done.Results (1)The success rate of glottis exposure,the one-time success rate and the success rate of tracheal intubation in group A-HC were 90%,70% and 90%,respectively which were significantly higher than those (50%,20% and 45%)in group A-N (P <0.05).The rate of complication,the number of attempt and operative time consumed for intubation cases in group A-HC were respectively 15%,(1.28 ± 0.43)and (31.44 ±5.06 ) s,which were significantly less than those [45%,(1.89 ±0.79 )and (45.89 ±4.99) s in group A-N (P <0.05 ).(2)The success rate of glottis exposure,the one-time success rate and the success rate of tracheal intubation in group B-N were 80%,65% and 80%, respectively which were significantly higher than those (50%,20% and 45%)in group A-N (P <0.05). The rate of complication,the number of attempt and operative time consumed for successful intubation cases in group B-N were 15%,(1.25 ±0.41)and (39.31 ±4.23)s,respectively which were significantly less than those [45%,(1.89 ±0.79)and (45.89 ±4.99)s]in group A-N (P <0.05).(3)There were no significant statistical differences in above results between group A-HC and group B-HC (P >0.05 ). Conclusions HC visual laryngoscope used by unskilled junior emergency resident doctors for emergency tracheal intubation,could increase success rate of glottis exposure,decrease the number of attempt of intubation,decrease rate of complication,shorten operative time required for intubation thus improving efficiency and success rate of emergency tracheal intubation,and maybe narrow the skill gap between unskilled junior resident doctors and well experienced attending doctors in respect of emergency tracheal intubation.
4.Effect of group cognitive behavioral therapy on blood glucose levels, anxiety and depression in patients with type 2 diabetes mellitus
Xiaobo GUAN ; Zheng LU ; Zhen GUO ; Tao LEI ; Xueli SUN ; Chengge GAO ; Dong GAO
Chinese Journal of General Practitioners 2015;14(3):206-211
Objective To evaluate the clinical effect of group cognitive behavioral therapy on blood glucose levels,anxiety and depression in patients with type2 diabetes mellitus.Methods Ninety three patients with type 2 diabetes mellitus were recruited and randomly divided into intervention group (n =48) and control group (n =45).Both groups received diabetes health education,patients in intervention group received additional group cognitive behavioral therapy.The glucose tolerance,glycosylated hemoglobin A1c were measured; the HAMA(Hamilton Anxiety Scale)scores,HAMD(Hamilton Depression Scale)scores and CSQ (Coping Styles Questionnaire) scores in patients were analyzed before and 6 months after treatment.Results After 6-month treatment the fasting blood glucose (6.33 mmol/L vs.5.94 mmol/L),1 h postprandial plasma glucose(12.40 mmol/L vs.11.46 rmool/L),2 h postprandial plasma glucose (10.24 mmol/L vs.9.13 mmol/L),A1 c (6.31% vs.6.07%) in intervention group were decreased significantly,compared to baseline values (all P < 0.05).The HAMA total score (9.98 vs.8.14),somatic anxiety (3.98 vs.3.48),psychic anxiety(6.00 vs.4.67),HAMD total score(10.74 vs.6.93),anxiety somatic(5.02 vs.3.26),block(2.24 vs.1.38)and sleep disorders(2.40 vs.1.40)in intervention group were all decreased significantly(P < 0.01 or 0.05).There were significant differences in HAMA total score (8.14 vs.9.15),HAMD total score(6.93 vs.9.33),anxiety somatic(3.26 vs.4.38),block(1.38 vs.1.98)and sleep disorders(1.40 vs.2.03)between the intervention group and control group(P < 0.01 or 0.05).And the negative coping style scores in intervention group was also lower than that of the baseline (26.74 vs..29.43).Conclusion The group cognitive behavioral therapy combined with diabetes health education for patients with type 2 diabetes mellitus may improve the glucose metabolism and depression and anxiety status of patients.
5.Influences of pancreatic cancer cell PANC-1 proliferation of downregulating Berlin1 expression
Xiaobo ZHANG ; Feng GAO ; Xiaodong TAN ; Lei ZHOU ; Huaitao WANG ; Gang SHI
Clinical Medicine of China 2015;31(8):677-681
Objective To observe the influences of Bcl-2 expression and pancreatic cancer cell proliferation after downregulating Beclin1 expression and discuss the mechanism.Methods Target interfering plasmid siBeclin1 was constructed and transfected to PANC-1 cell,and cells were divided into non transfected group(Beclin1 group),blank control group(control siNegative group) and transfection group(siBeclin1 group).Western blot and qRT-PCR were used to detect the transfection efficiency and Bcl-2 expression after transfection.Proliferation was detected by MTT.Results Western blot and qRT-PCR results showed that expression of Beclin1 in siBeclin1 group was lower than Beclin1 and siNegative control groups (27.823± 1.432,8.635±1.481,26.904±1.098;F=8.176,P<0.01),qRT-PCR showed that the Beclin1 mRNA level of PANC-1 cells in siBeclin1 group was lower than Belcin1 and siNegative groups (0.421 ±0.157,0.194±0.104,0.399 ±0.123;F=5.239,P<0.01),and the silencing rate was about 60%-70%.Bcl-2 protein expression and levels of mRNA in siBeclin1 group were significantly higher than that of Beclin1 group(26.912±1.927,8.004±1.534,t =7.329,P<0.01;0.582±0.297,0.217±0.186,t =6.835,P<0.01);MTT results showed that the proliferation of PANC-1 cells were strengthened,(61.54±6.81)% and (46.78±7.28)% at 72 h,(76.39±7.26)% and (54.27±8.17)% at 96 h,and the difference was significant(t=3.674,P<0.05;t =10.185,P <0.01).Conclusion Proliferation of pancreatic cancer cells are strengthened after Beclin1 expression downregulating,so Beclin1 may inhibit the proliferation of pancreatic cancer cells by Bcl-2 downregulating.Beclin1 can be a target candidate gene of pancreatic cancer therapy.
6.Spiral CT Findings of Intrahepatic Spontaneous Portosystemic Venous Shunts in Liver Cirrhosis
Huaming WANG ; Xiaobo LIU ; Rong ZHAO ; Rujin WANG ; Lei GAO ; Hui XIE
Journal of Practical Radiology 2001;0(01):-
Objective To describe CT manifestations of intrahepatic spontaneous portosystemic shunts(ISPS) of cirrhosis.Methods Spiral CT(SCT) scans and related clinical findings of 15 cirrhotic patients with ISPS were retrospectively reviewed.Results Three kinds of ISPS were showed :①Right posterior portal vein type was in 12 cases and showed tortuous intrahepatic tubular structures which were all in the posterior segment of the right lobe connecting the right posterior portal vein to the inferior vena cava(IVC) in the suprarenal region in all cases.②Hepatic venous type was in 2 cases,the portal vein communication with the hepatic vein in the liver.③Apex type was in 1 case,the vein arose from the apex of the liver and drains into the internal thoracic vein.Blood ammonia levels were elevated in all cases and symptoms associated with hepatic encephalopahty were present in 2 cases.Conclusion To deeply understand the ISPS,it is of benefit for differential diagnosis of focal hepatic lesions.
7.(2009ZC116M).Relationship of biomaterials related infections and transforming growth factor levels in the peripheral blood of lung cancer patients
Yujie LEI ; Yunchao HUANG ; Li YANG ; Fengli GUO ; Guangqiang ZHAO ; Yushan XU ; Xiaobo CHEN ; Kaiyun YANG
Chinese Journal of Tissue Engineering Research 2011;15(47):8907-8910
BACKGROUND: Permanent or transient implantation of biomaterials can result in biomaterials-centered infections (BCI) in lung cancer patients.OBJECTIVE: To investigate the relationship between BCI and peripheral blood transforming growth factor β1 (TGF-β1) in patients with lung cancer.METHODS: A total of 248 lung cancer patients undergoing in vivo intravascular catheter indwelling > 7 days were included.Quantitative method was used for intubation, bacteriological culture and paired blood culture, and API Staph strips were adopted for positive patients. While enzyme-linked immunosorbent assay was used to detect TGF-β1 levels in the peripheral blood of patients with lung cancer and 75 healthy volunteers as normal controls.RESULTS AND CONCLUSION: Among the 248 patients, there were 82 BCI-positive cases, and 166 BCI-negative cases.Thirteen patients were confirmed to have catheter-related bloodstream infection. There were 48 Gram-positive bacteria, 24Gram-negative bacilli, and 10 fungal. The levels of TGF-β1 were higher in BCI-positive patients than BCI-negative patients (P < 0.05); the levels of TGF-β1 in the BCI-negative group were higher than those in the normal control group (P < 0.05). For lung cancer patients with nosocomial infection induced BCI, there are various species of pathogenic bacteria, and Gram-positive bacteria are more common. To detect TGF-β1 levels in patients with lung cancer is of significance for early prevention of BCI.
8.Rapid identification and sequence typing of genotype VG Ⅱ of Cryptococcus gattii
Xiaobo FENG ; Bo LING ; Xiaohua FU ; Lei WANG ; Daming REN ; Zhirong YAO
Chinese Journal of Clinical Infectious Diseases 2012;05(1):1-4
Objective To establish a PCR method for rapid identification and sequence typing of VG Ⅱ allele of the intergenic spacer region (IGS) of Cryptococcus gattii.Methods Since IGS1 was of high sequence variation,multiple alignments were conducted by ClustalX 2 in IGS1 of Cryptococcus gattii and Cryptococcus neoformans,and then primer sets specific to genotype VG Ⅱ was designed for PCR analysis.The specificity of the primer pair was detected by amplification of the other genotypes in Cryptococcus gattii,Cryptococcus neoformans,and other pathogenic yeasts.The amplified fragments from VG Ⅱ genotype were sequenced and subtyped.Results Using the PCR analysis developed in this study,all VG Ⅱ genotype strains tested were amplified,whereas no amplification was obtained from other genotypes or yeast species involved herein.Three polymorphic nucleotide sites at 72,79 and 104 bp in the fragment amplified could be used to distinguish sub-genotypes within VG Ⅱ genotype.Conclusions The PCR analysis developed in this study can be used for rapid identification of genotype VG Ⅱ of Cryptococcus gattii.The sequence typing based on the amplified fragment from IGS1 may be performed for screening the highly virulent sub-genotype VGⅡ a.
9.Identification of Cryptococcus neoformans var.grubii, var.neoformans and Cryptococcus gattii by multiplex PCR
Xiaobo FENG ; Bo LING ; Xiaohua FU ; Lei WANG ; Daming REN ; Zhirong YAO
Chinese Journal of Dermatology 2012;(12):870-873
Objective To establish a multiplex PCR targeting the intergenic spacer regions (IGS) for the identification of Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii.Methods Primers were designed by using the software ClustalW2 and Oligo 6 based on the sequence of IGS1 region,which shows high sequence variability in the genome of Cryptococcus neoformans and Cryptococcus gattii.,for the multiplex PCR.Then,the developed multiplex PCR was performed to identify 51 Cryptococcus neoformans strains representing genotypes VNI-VNIV and VNB as well as 41 Cryptococcus gattii strains representing genotypes VGI-VGIV.The identification results were compared with those from common PCR by using primers GPA1A,CLA4D and SODlgattii specific to Cryptococcus neoformans var.grubii,var.neoformans and Cryptococcus gattii,respectively,as well as with those from the canavanine-glycine-bromothymol blue (CGB) medium-based culture.Results The developed multiplex PCR successfully identified the 92 Cryptococcus neoformans and Cryptococcus gattii.strains,and yielded negative results from the other tested pathogenic yeasts,which revealed a high specificity of the designed primers.False positive results were observed in the identification of two Cryptococcus gattii strains with GPA1A primer-based PCR,one Cryptococcus gattii strain with CLA4D primer-based PCR,one var.grubii strain and one var.neoformans strain with CGB culture,while no false negative results were observed in the detection of these Cryptococcus strains by any of these methods.Conclusions The developed multiplex PCR in this study can rapidly and accurately identify Cryptococcus neoformans var.grubii,var.neoformans,AD hybrid,and Cryptococcus gattii,with superior performance in comparison with common PCR and CGB medium-based culture.
10.Endoscopic submucosal dissection for tumors originating from gastric muscularis propria
Chenghong FU ; Hanbing XUE ; Xiaobo LI ; Lei SHEN ; Jie ZHUANG ; Hui CAO ; Zhizheng GE
Chinese Journal of Digestive Endoscopy 2012;29(7):385-388
Objective To explore the feasibility,efficacy and safety of endoscopic submucosal dissection (ESD) for tumors originating from gastric muscularis propria.Methods A total of 20 patients with tumors originating from gastric muscularis propria confirmed by EUS and CT scan underwent ESD.Laparoscopic intervention was applied in 3 cases when difficulties in en bloc removal or perforation occurred.Results The mean ESD procedure time was 74.8 min (40-120 min),and the mean resected tumor size was 2.6 cm (1.5-3.5 cm).No severe complication occurred.Pathological findings were 17 cases of gastrointestinal stromal tumors and 3 cases of leiomyoma.Conclusion ESD is an effective method for treating the tumors originating from gastric muscularis propria,and laparoscopic intervention is necessary for en bloc resection in some cases.