3.Effects of lentivirus-mediated heat shock protein 70 gene on endoplasmic reticulum calcium homeostasis and calcium channels of PC12 cells induced by ischemia and hypoxia
Yuan LIU ; Chun GUAN ; Lulu GUO ; Qingshu LI ; Yun WANG ; Chunyu XIE ; Dan HU ; Yan QU
Chinese Critical Care Medicine 2016;28(3):205-210
Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on calcium homeostasis and calcium channels of PC12 cells induced by ischemic and hypoxia and its mechanisms. Methods PC12 cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing HSP70 and green fluorescent protein (GFP) fluorescin gene], lentivirus control group (infected by lentivirus containing GFP without HSP70 gene) and non-infection group. PC12 cells were subjected ischemia/hypoxia for 4, 8, 12, 24 hours, and the cell activity was determined by methylthiazolyl tetrazolium (MTT) assay test inorder to determine the best time for ischemia/hypoxia. The mRNA expressions of HSP70, muscle/endoplasmic reticulum Ca2+-ATP isoforms (SERCA2a, SERCA2b), ryanodine receptor 2 (RyR2), and inositol 1,4,5-triphosphate receptor 1 (IP3R1) were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expressions of HSP70, SERCA, and IP3R were determined by Western Blot at 8 hours after ischemic/hypoxia. Flow cytometry was used to determine the levels of intracellular reactive oxygen (ROS) and intracellular Ca2+ ([Ca2+]i). Results With the prolongation of time of ischemia/hypoxia, the cell viability in all groups showed an increase followed by a weakening, and peaked at 8 hours. The cell viability at 8 hours in lentiviral infection group was significantly higher than that of the non-infection group and lentivirus control group [A value (×10-2): 20.3±2.2 vs. 14.1±2.1, 15.0±1.6, both P < 0.01], the mRNA and protein expressions of HSP70 and SERCA in lentiviral infection group were significantly increased [HSP70 mRNA (2-ΔΔCt ): 0.785±0.018 vs. 0.428±0.019, 0.423±0.023; HSP70 protein (gray value): 2.72±0.20 vs. 1.56±0.36, 1.63±0.41; SERCA2a mRNA (2-ΔΔCt ): 0.971±0.037 vs. 0.367±0.014, 0.347±0.012; SERCA2b mRNA (2-ΔΔCt ): 8.869±0.162 vs. 3.015±0.091, 2.941±0.091; SERCA protein (gray value): 2.84±0.18 vs. 1.48±0.26, 1.52±0.29], and IP3R2 mRNA and protein expressions were significantly declined [IP3R2 mRNA (2-ΔΔCt ): 0.183±0.020 vs. 0.439±0.020, 0.433±0.040; IP3R2 protein (gray value): 1.15±0.12 vs. 1.91±0.20, 1.83±0.19], with statistically significant differences (all P < 0.01); no significant difference in RyR mRNA was found [2-ΔΔCt (×10-3): 1.97±0.63 vs. 2.02±0.22, 2.01±0.09, both P > 0.05]; the relative fluorescence intensity of ROS and [Ca2+]i in lentiviral infection group was significantly reduced (ROS: 30.54±1.23 vs. 58.03±1.97, 57.72±2.35; [Ca2+]i: 34.50±2.05 vs. 48.20±3.02, 46.80±2.75, all P < 0.01]. Conclusion Exogenous HSP70 can maintain calcium homeostasis in the endoplasmic reticulum of PC12 cells, affect the Ca2+ channel protein regulated by calcium channel IP3R and calcium pump SERCA, which may cause hypoxia/ischemia intracellular injury.
4.Effect of Paroxetine on Post-stroke Depression
Ru-qin LUO ; Chun-hua HU ; Ren-yun CHEN ; Anqi WANG ; Yusheng REN
Chinese Journal of Rehabilitation Theory and Practice 2006;12(7):597-598
ObjectiveTo observe the efficacy and side effects of paroxetine and amitriptyline in the treatment of post-stroke depression (PSD).Methods82 PSD patients were randomly divided into the paroxetine group (n=42) and amitriptyline group (n=40). Before the treatment and at the ends of 2nd, 4th and 8th week after the treatment, all patients in two groups were assessed with Hamilton Depression Scale (HAMD) and TESS.ResultsAfter treatment, HAMD scores of patients of two groups were decreased significantly (P<0.01), but no difference was found between two groups. The side effects of the paroxetine group were obviously fewer and milder than that of amitriptyline group (P<0.01).ConclusionParoxetine is an effective and safe drug in the treatment of PSD.
5.Anatomic measurements and quantitative analysis of posterior acetabular wall.
Yang TANG ; Yun-tong ZHANG ; Chun-cai ZHANG ; Pan-feng WANG ; Xue ZHAO ; Xiong-wei LU ; Xiao-peng HU
China Journal of Orthopaedics and Traumatology 2014;27(12):1024-1028
OBJECTIVETo explore morphological character and clinical significance of superior-posterior acetabular wall by anatomically measuring and quantitatively analyzing thickness of posterior acetabular wall, then provide a theoretical reference for clinical treatment of acetabular fracture.
METHODSFifteen adult formalin-preserved cadaveric pelvises (8 males and 7 females) were used for this investigation. Excess soft tissue was removed and the whole acetabular posterior walls were marked with "angle" sector method and the thickness was measured with caliper in different levels of the different split points. The measurement results were validated and analyzed statistically.
RESULTSAt 5 mm away from acetabular rim, the average thickness of superior-posterior acetablar wall fluctuated between (6.47±0.61) mm and (7.43±0.71) mm; the average thickness of inferior-posterior acetabuluar wall fluctuated between (5.62±0.51) mm and (6.33±0.61) mm; the average thickness of acetabular roof fluctuated between (7.71±0.74) mm and (8.27±0.99) mm. There was no statistical difference between average thickness of superior-posterior wall of acetabulum and inferior-posterior wall of acetabulum (P>0.05), but the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.05). At 10 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (8.81±0.67) mm and (13.35±0.89)mm; the average thickness of inferior-posterior acetabular wall fluctuated between (7.02±0.63) mm and (7.66±0.69) mm; the average thickness of acetabular roof fluctuated between (14.46±0.97) mm and (17.05±1.35) mm. Comparatively, the average thickness of superior-posterior acetabular wall was significantly larger than inferior-posterior wall of acetabulum (P<0.05), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P<0.01). At 15 mm away from the acetabular rim, the average thickness of superior-posterior acetabular wall fluctuated between (12.08±0.78) mm and (19.84±1.03) mm; the average thickness of inferior-posterior acetabular wall fluctuated between (10.17±0.76) mm and (11.12± 0.77) mm; the average thickness of acetabular roof fluctuated between (23.23±1.12) mm and (26.01±1.53) mm. Comparatively, the average thickness of superior-posterior wall of acetabulum was significantly larger than inferior-posterior acetabular wall (P<0.01), and the average thickness of acetabular roof was significantly larger than superior-posterior acetabular wall (P< 0.01).
CONCLUSIONThe thickness of entire acetabular posterior edge revealed an increasing tendency from inferior-posterior wall to the superior-posterior wall to acetabular roof. And this trend became more obvious with increasing distance away from acetabular rim. Therefore, the superior-posterior acetabular wall could not only maintain the stability of hip joint but also bear loading.
Acetabulum ; anatomy & histology ; injuries ; surgery ; Female ; Humans ; Male
7.Diagnostic study on children's diaphragmatic fatigue.
Yun-gang YANG ; Zheng-xia ZHANG ; Liang-gang HU ; Chun-xue YAN
Chinese Journal of Pediatrics 2003;41(4):274-277
OBJECTIVEIn the recent twenty years, the diaphragmatic contraction, relaxation functions and electric activity have been explored through electromyography (EMG) and transdiaphragmatic pressure (Pdi) determination. But these techniques required some complex and expensive instruments, so the diagnosis and treatment of children's diaphragmatic fatigue have not been well evaluated. The present study explored the diagnosis of children's diaphragmatic fatigue through measuring ribcage-abdomen motion and analyzed its asynchrony.
METHODSFifty-three children (male 37, female 16, and age rage from 1 months to 9 years) with respiratory rate > 30 breaths/min, heart rate > 110 beats/min, and respiratory dysfunction had asynchronized ribcage-abdomen motion showed by impedance respirograph (IRG). The authors observed whether ribcage-abdomen motion was synchronic and calculated M levels (staggered peak time/total duration of the breathing cycle). The ribcage and abdomen outputs were displayed on vertical (for rib cage) and horizontal (for abdomen) axes of X-Y instrument. In addition, the change of respiratory frequency and heart rate was observed and arterial blood-gas analysis was also performed.
RESULTS(1) M levels in one-dimensional IRG were positively correlated with alpha angle in two-dimensional IRG (r = 0.956, P < 0.001). Asynchronized respiratory motions could be divided into three types. type I showed completely contra-directional movements of respiration, M levels for (48.1 +/- 4.4)%, an irregularly clockwise loop in the two dimensional IRG, and alpha angle for (138.3 +/- 15.0) degrees. In type II, one dimensional IRG showed displaced peak of the chest and abdomen motion curves, M levels were (16.5 +/- 4.7)%, two dimensional IRG was displaced in a counterclockwise direction, and alpha angle was (55.3 +/- 10.8) degrees. In type III, abdominal motion curve of one dimensional IRG had double peaks, M levels were 0, two dimensional IRG was presented as 8-shaped double circles, alpha angle was (41.3 +/- 3.8) degrees; (2) pH levels in the patients with type I and type II diaphragmatic fatigue were significantly lower, and PCO(2) levels were significantly higher than those with type III or in the normal subjects (P < 0.001 for all), but there was no statistically significant difference between type III and the normal subjects (P > 0.05); (3) Both of respiratory rate and heart rate in type I, type II and type III were higher than those in the normal subjects (all P < 0.001), and the differences among the three types were significant (P < 0.001 for all); (4) Both M levels and alpha angle were negatively correlated with pH levels (r = -0.514, P < 0.001 and r = -0.497, P < 0.001), while positively correlated with PCO(2) levels (r = 0.672, P < 0.001 and r = 0.625, P = 0.01).
CONCLUSIONS(1) IRG can be reliably used to diagnose children's diaphragmatic fatigue. This technique is simple and easy to perform and non-invasive. It is therefore worthy of recommending for further clinical investigations. (2) According to the characteristics of IRG, diaphragmatic fatigue can be divided into three types. (3) The development of children's diaphragmatic fatigue has a series of characteristic changes. (4) To avoid the patients suffering from respiratory failure, it is the key time to adopt the policies of prevention and treatment when IRG shows signs of type III diaphragmatic fatigue.
Child ; Child, Preschool ; Diaphragm ; physiopathology ; Fatigue ; classification ; diagnosis ; Female ; Humans ; Infant ; Male ; Respiration ; Respiratory Function Tests ; methods
8.Surgical correction of male external genitalia giant condyloma acuminata.
Xiao-yun PENG ; Zhong-dai YUAN ; Chun-mei HU ; Xing YUAN
Chinese Journal of Plastic Surgery 2007;23(4):304-306
OBJECTIVETo observe the clinical efficiency of curettage combining circumcision on giant condyloma acuminata(CA) in male external genitalia, the relationship between recurrence and curettage depth, the possibility of HPV infection in PBMC after operation.
METHODSCurettage combining circumcision was carried out on 50 cases with CA. The removed wart and wound surface tissues were examined under light microcope and for HPV-DNA detection by PCR. HPV-DNA was detected in PBMC during < or =1 and > or =2 weeks after operation.
RESULTS(1) 46 cases were cured completely after one treating (once the cure rate is 92%), 4 cases twice(twice the cure rate is 8%); (2) The tissues were proved to be HPV-DNA positive by PCR amplification and CA relapse occurred; (3) HPV-DNA was detected in PBMC only within the week after curettage in the 19 cases.
CONCLUSIONS(1) The clinical efficiency of curettage combining circumcision on giant condyloma acuminata in male external genitalia is sure; (2) Cure rate and relapse rate are related with curettage depth; (3) Transient positive HPV-DNA in PBMC may be detected.
Adolescent ; Adult ; Aged ; Circumcision, Male ; Condylomata Acuminata ; surgery ; Curettage ; methods ; Humans ; Male ; Middle Aged ; Young Adult
9.Glomus tumor of the gastric body: helical CT findings.
Xiao-Yun HU ; Chun-Hong HU ; Xiang-Ming FANG ; Tong-Hua ZHANG
Chinese Medical Journal 2007;120(14):1289-1291
10.Anti-tumor effect and mechanism of Paeonol on the hepatocellular carcinoma cell line Bel-7404.
Chun-hu ZHANG ; Sui-yu HU ; Yun-hui LI ; Mei-qun CAO
Journal of Central South University(Medical Sciences) 2006;31(5):682-695
OBJECTIVE:
To investigate the anti-tumor effect of Paeonol (Pae) on the hepatocellular carcinoma cell line Bel-7404 and its molecular mechanisms.
METHODS:
Hepatocellular carcinoma cell line Bel-7404 was treated by Pae in various concentrations and different time points respectively; and then the cell proliferation was assayed by light microscope, MTT method. DNA agarose gel electrophoresis and TUNEL were used to detect the apoptosis. The expression of PTEN and Akt were examined by RT-PCR and immunocytochemical ABC method.
RESULTS:
Compared with the control groups Pae obviously increased the inhibitory and apoptosis rate of hepatocellular carcinoma cell line Bel-7404. It also showed a typical apoptotic morphology and DNA depicted a ladder pattern characteristic of the apoptosis, indicating the presence of DNA fragmentation. RT-PCR and immunocytochemical ABC assay showed that Pae could increase the expression of PTEN and decrease the expression of Akt.
CONCLUSION
Pae can increase the anti-hepatocellular carcinoma effect, and its mechanism may be the increase of apoptosis-inducing effect which is regulated by phosphatidylinositol-3-kinase.
Acetophenones
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pharmacology
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Antineoplastic Agents, Phytogenic
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pharmacology
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Apoptosis
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drug effects
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Carcinoma, Hepatocellular
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pathology
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Cell Proliferation
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drug effects
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Humans
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Liver Neoplasms
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pathology
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Phosphatidylinositol 3-Kinases
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metabolism
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Tumor Cells, Cultured