1.PKH26 combined with Hoechst 33258 to trace the iRhom2 gene and its mutant proteins of Uncv mice in Vero cells
Ying YOU ; Bingbo CHEN ; Lin ZENG
Chinese Journal of Comparative Medicine 2017;27(8):40-42
Objective To determine the localization of iRhom2 and its mutant proteins of Uncv mice in Vero cells by PKH26 combined with Hoechst 33258 staining.Methods The cell membrane was stained with PKH26, and the nuclei were stained with Hoechst 33258 dye, and observed by laser scanning confocal microscopy.Results It was found that wild iRhom2 was distributed in the cytoplasm, and its iRhom2mut was present both in cytoplasm and cell nuclei.Conclusions The results of our study suggest that a deletion in N-terminal of iRhom2 affects its subcellular localization.
2.Association between the efficacy of alendronate and serum level of vitamin D in patients with osteoporosis
Ling PAN ; Li YOU ; Yongde PENG ; Jinyu CHEN ; Lin CHEN
Chinese Journal of Endocrinology and Metabolism 2014;30(4):312-315
Objective To examine the association between vitamin D and alendronate response,and to investigate the proper vitamin D levels for the efficacy of alendronate treatment of osteoporosis.Methods In this retrospective study,559 post-menopausal women with osteoporosis were devided into two groups:non-responders and responders,based on the European Study of Forsteo (EUROFORS).Demographic and clinical data including mean 25-hydroxy vitamin D (25-OHD) levels between dual-energy X-ray absorptiometry (DEXA) scans were obtained.Mean 25-OHD levels were compared between responders and non-responders and multiple logistic regression analysis was performed to identify factors associated with non-response.Results There were 437 (75.5%) responsers to alendronate therapy and 142 (24.5%) non-responders.Responders with a mean serum 25-OHD level of (59.96 ±12.56) nmol/L,obtained a more favorable result than non-responders,whose serum 25-OHD level was (47.50 ±9.92) nmol/L (P<0.01).25-OHD level was significantly associated with response.Conclusion Patients with a mean 25-OHD ≥50 nmol/L had a substantially greater likelihood of maintaining bisphosphonate response.
3.Relation of body mass index to vitamin D, PTH, and bone turnover markers levels among women in Shanghai area
Li YOU ; Lin CHEN ; Ling PAN ; Jinyu CHEN ; Yongde PENG
Chinese Journal of Endocrinology and Metabolism 2013;29(7):566-569
Objective To examine the relationship of body mass index (BMI) with levels of vitamin D,parathyroid hormone (PTH),and bone turnover markers among women in Shanghai area.Methods Altogether 810 Chinese women aged 45 year and older were enrolled to study the associations by multiple linear regression analyses.Bone mineral density was measured by a dual energy X-ray absorptiometry (Lunar,Prodigy,GE),Serum 25-hydroxyvitamin D [25 (OH) D],PTH,C-telopeptide of type Ⅰ collagen (CTX),osteocalcin (BGP),and bonespecific alkaline phosphatase (BSALP) were measured.Results (1) 25 (OH) D,BGP,BSALP,and CTX levels were significantly lower in obesity group than in control group ; PTH level was significantly higher in obesity group than in control group.(2) In multiple linear regression analyses,BMI was significantly associated with lower 25 (OH) D (β =-0.017,P =0.006),BGP (β =-0.077,P =0.019),and higher PTH (β =0.025,P =0.000).Conclusions (1) There were siginificant associations between higher BMI and lower 25 (OH) D among women in Shanghai.(2) Serum osteocalcin was associated not only with bone metabolism but also with energy metabolism.
4.Beta-amyloid affects retinoid receptor-α exporting into cytoplasm
Xiaoqing YOU ; Yaomin CHEN ; Xiumei HUANG ; Yingchun LIU ; Jianyin LIN
Chinese Journal of Neurology 2011;44(10):706-710
Objective To investigate the effect of Alzheimer' s beta-amyloid (Aβ) on the production and the translocation in cytoplasm of retinoid receptor-α (RXRα). MethodsN2awt cells were treated with Aβ peptide or amyloid protein precursor(APP695) transfection. The nucleus were separated from the cytoplasm by kit. The quantity of RXRα in the nucleus and cytoplasm was detected by Westernblot. The translocation of RXRα in the nucleus and cytoplasm of above N2awt cells or of the cortex cells in the brains of Alzheimer' s disease (AD) patients and their normal control groups was observed by immune fluorescence. ResultsIn N2awt cells, the increasing APP or Aβ had no significant effect on the production of RXRα but resulted in RXRα exporting into cytoplasm, the ratio of RXRα in cytoplasm increased from 3.2% (in control group) to 17.6% (in APP+ group) and from 3.8% (in control group) to 14.3% (in Aβ + group) respectively; compared with normal cortex cells, the translocation of RXRα in the cytoplasm of the cortex cells in the brains of AD increased significantly. ConclusionAβ may affect RXRα exporting into cytoplasm.
5.Effect of GSK-3βtargeting RNAi recombinant adenovirus on proliferation of human umbilical vein endothelial cells
Gang CHEN ; Tingting YOU ; Yufang QIAO ; Xiaoyan SHEN ; Lixiang LIN
Chinese Journal of Endocrinology and Metabolism 2008;24(6):654-657
Objective To observe the effect of Wnt/β-catenin pathway on proliferation of human umbilical vein endothelial cell (HUVEC) using the glycogen synthase kinase 3β (GSK-3β)-targeting RNAi recombinant adenovirus vector. Methods Homologous recombination and cloning techniques were used to construct RNAi recombinant adenoviral expressive vectors specific to GSK-3β. Then, the adenovirus plasmids was transfected into HEK 293A cells to produce adenovirus and amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. The GSK-3β and β-catenin protein expressions were detected by Western blot and immunohistochemistry. The proliferation of HUVEC was detected with MTr assay. Results The RNAi adenovirus vectors specific to GSK-3β were successfully produced with high titer. The expression of GSK-3β protein in HUVEC could be down-regulated efficiently by the RNAi adenovirus, along with increased β-catenin protein expression. The proliferation of HUVEC was significantly increased (P < 0.05 or P < 0.01) after infected with GSK-3β RNAi recombinant adenovirus for 3, 5, 7 days. Conclusion RNAi adenovirus is an important tool that can inhibit the expression of GSK-3β efficiently, along with increased β-catenin protein expression. Up-regulating of the Wnt/β-catenin pathway might play an important role in the proliferation of HUVEC.
6.Expression of angiotensin Ⅱ and its type 1 receptor in circulation, placenta and kidney of the preeclampsia rat model
Lin TAO ; Jianying NIU ; Li YOU ; Jing CHEN ; Yong GU
Chinese Journal of Nephrology 2008;24(9):663-668
Objective To investigate the expression of angiotensinⅡ (Ang Ⅱ ) and its type 1 receptor (AT1) in circulation, placenta and kidney of the rots preeclampsia. Methods Preedampsia rat model was developed by inhibitor of nitric oxide synthase (L-NAME). The systolic blood pressure (SBP), 24 h urine protein, hepatic and renal function were compared among the precelampsia group, the normal pregnant group and nonpregnant control group. The kidney tissue was observed by light microscopy. ELISA and radioimmunoassay were used to detect Ang Ⅱ in rat plasma and kidney homogenate respectively. Placental AT1 was measured by Westem blot. The level of kidney AT1 was evaluated by immunohistochemistry and Western blot. Results In preeclampsia rats, SBP and 24 h urine protein were significantly higher compared with control groups. Compared to normal pregnant group, plasma Ang Ⅱ of preeclampsia rats was much higher [(0.706±0.086) ng/L vs (0.540±0.085) ng/L, P<0.05]; placental AT1 was increased by 46%(P< 0.05); kidney Ang Ⅱ was decreased signigicantly [(65.543±40.634) ng/g vs (165.543±33.078) ng/g, P<0.05]. The expression of ATI in kidney of preeclampsia rats was reduced evidently,which was only 33% of normal pregnancy group and 59% of nonpregnant control greup,respeetively (P<0.05). Conclusions In preeclampsia rat model, the circulating Ang Ⅱ is increased, the placental RAS isactivated, while the kidney BAS is suppressed. The underlying mechanism of proteinuria and kidney damage associated with this phenomenon in preeclampsia needs further research.
7.Value of 18F-FDG PET/CT in the post-treatment evaluation of cervical cancer
Feng CHEN ; Tianzeng LIN ; Chunjing YU ; Xuyang YOU ; Weixing WAN
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(3):164-167
Objective To evaluate the value of whole body 18F-FDG PET/CT in the detection of recurrence/metastasis of cervical cancer.Methods A retrospective study was performed on 95 patients with cervical cancer who underwent 18F-FDG PET/CT after treatment.The lesion characteristics on 18F-FDG PET/CT were assessed visually and semi-quantitatively.A final diagnosis was confirmed by histopathology,diagnostic treatment and clinical follow-up imaging.The data were analyzed by Kappa test.Results 18 F-FDG PET/CT was positive in 54 patients,including 24 with local recurrence and 30 with distant metastases.The sensitivity,specificity and accuracy of 18F-FDG PET/CT for the detection of recurrence/metastasis of cervical cancer were 98.1% (52/53),95.2% (40/42) and 96.8% (92/95),respectively.The positive predictive and negative predictive value were 96.3% (52/54) and 97.6% (40/41),respectively.18F-FDG PET/CT showed concordant results with pathological/clinical follow-up findings (Kappa =0.936,P<0.05).Conclusion 18F-FDG PET/CT is a sensitive and specific modality for the detection of recurrence/metastasis of cervical cancer and might be useful for further treatment plan.
8.Role of fibroblast growth factor-2 in neuropathic pain in rats
Jiyue YOU ; Jin GAO ; Ping CHEN ; Lin SUN ; Xiaoqiu YANG
Chinese Journal of Anesthesiology 2012;(9):1108-1110
Objective To evaluate the role of fibroblast growth faetor-2 (FGF-2) in neuropathic pain (NP) in rats.Methods One hundred male Sprague-Dawley rats,aged 7 weeks,weighing 200-250 g,were randomly divided into 4 groups (n=25 each): sham operation group (group S),group NP,phosphate buffer solution (PBS) group and FGF-2 antibody group (group Ab).FGF-2 antibody 18 μg (40 μl) was injected intrathecally at 1,6,9,13,16 and 20 days after operation in group Ab,while the equal volume of PBS was injected intrathecally in group PBS.Spinal nerve injury was produced by exposing the sciatic nerve and its branches and ligation and transection of tibial nerve and common fibular nerve.Pain behavior was assessed at 1 day before operation and at 1,4,7,14 and 21 days after operation.The paw withdrawal mechanical threshold (PWMT) was measured.The animals were then sacrificed and the lumbar segment (L4-6) of the spinal cord was obtained for determination of the contents of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the spinal cord.Results Compared with group S,the PWMT was significantly decreased and the contents of TNF-α and IL-6 in the spinal cord was significantly increased in groups NP,PBS and Ab (P < 0.05).The PWMT was significantly higher and the contents of TNF-α and IL-6 in the spinal cord were significantly lower in group Ab than in groups NP and PBS (P < 0.05).Conclusion FGF-2 is involved in the occurrence and development of NP and induction of the inflammatory response in the rat spinal cord in involved in the mechanism.
9.The relationship between soluble tumor necrosis factor receptor and insulin resistance
Jie YOU ; Yao CHEN ; Peiji HUANG ; Zhezhang LIN
Chinese Journal of Endocrinology and Metabolism 1985;0(01):-
Objective To study the relationship between soluble tumor necrosis factor receptor (STNFR) 1,2 and insulin resistance (IR). Methods STNFR1 and STNFR2 were measured by ELISA in 43 men and 41 premenopausal women. IR was assessed by Homa Model. Results Obese men and women showed higher levels of STNFR2 than nonobese men and women (P
10.Drug resistance and serological detection of Candida isolated from a fungal surveillance network in Hainan province from 2013 to 2022
WANG Like ; CHEN Xiaojuan ; ZHANG You ; CHEN Lin ; LIN Chong ; PEI Hua
China Tropical Medicine 2024;24(2):136-
Objective To collect data of fungi isolated from a fungus monitoring network in Hainan Province from 2013 to 2022, and analyze the drug resistance characteristics of Candida and the results of serological tests, with an aim to provide a basis for clinical diagnosis and treatment. Methods In accordance with the National Fungal Drug Resistance Monitoring Network technical scheme, the qualifying fungal data were extracted from the microbial identification system database using SQL language, and the data information was then analyzed, with statistical processing done using SPSS 26.0 software. Results Among 5 503 fungal isolates from clinical specimens between 2013 and 2022, cervical orifice secretions accounted for 30.37%(1 671 strains), mid-stream urine for 23.55%(1 296 strains), lower respiratory tract specimens for 25.24% (1 389 strains)[(sputum for 20.37%(1 121 strains) and alveolar lavage fluid for 4.87%(268 strains)], wound pus for 9.59%(528 strains), ascites for 5.60%(308 strains), blood for 3.67%(202 strains), cerebrospinal fluid for 0.38%(21 strains), and joint fluid for 0.04%(2 strains), with the highest number of strains isolated in 2022 and the lowest in 2013, the 2022 figure is about 2.6 times that of 2013. Among yeast-like fungi, Candida albicans had the highest proportion with 3 312 strains accounting for 60.2%; The highest resistance rate of Candida albicans was to fluconazole at 16.7%, with 2.5% being non-wild type (NWT) for amphotericin B; Candida tropicalis had the highest rate of resistance to fluconazole at 36.0%, with NWT at 41.1% for fluconazole and 3.1% for amphotericin B; Candida glabrata had a resistance rate to fluconazole of 2.8%, dose-dependent susceptibility (SDD) of 97.2%, NWT of 15.5% for fluconazole, and NWT of 8.6% for itraconazole; Candida parapsilosis had the highest resistance rate to fluconazole at 15.7% and and NWT of 8.3% for amphotericin B; Candida krusei had a 0.0% resistance rate to caspofungin; and Candida dubliniensis was 100.0% NWT to fluconazole. Of 70 cases of blood culture-positive specimens, 64 cases were detected by G test and 25 cases by Mn test, and the positive blood cultures were statistically significant when compared with the G test and Mn test, respectively (P<0.01). Conclusions Fungal serological test can make up for the deficiency of blood culture and distinguish fungal invasion and colonization, thus providing a basis for the effective control of fungal infection in clinical practice.