1. Expression and function of HSP110 family in mouse testis after vasectomy
Asian Journal of Andrology 2017;19(3):355-361
HSP110 functions to protect cells, tissues, and organs from noxious conditions. Vasectomy induces apoptosis in the testis; however, little is known about the reason leading to this outcome. The aim of the present study was to evaluate the expression and function of HSP110 in mouse testis after vasectomy. Following bilateral vasectomy, we used fluorescent Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to detect apoptosis, Western blotting and immunohistochemistry to examine HSP110 expression and localization. Serum antisperm antibody (AsAb) and testosterone were measured by Enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay, respectively. Expression of endoplasmic reticulum stress (ERS) sensors and downstream signaling components was measured by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and the phosphorylation of eIF2 and JNK was detected by Western blotting. Vasectomy induced morphologic changes, increased apoptosis in the testis, increased serum AsAb, and decreased testosterone levels. After vasectomy, ORP150 mRNA level was increased first and then decreased, Bcl-2 was decreased, and the expression of HSPA4l, GRP78, GADD153, PERK, ATF6, IRE-1, XBP-1s, Bax, Bak, and caspases and the phosphorylation of eIF2 and JNK were increased. We present that an ER stress-mediated pathway is activated and involved in apoptosis in the testis after vasectomy. HSPA4l and ORP150 may play important roles in maintaining the normal structure and function of testis.
2.Quantitative analysis of SOX9 and type Ⅱ collagen mRNA in the three-lineage differentiation of rat mesenchymal stem cells
Dachuan LIANG ; Jieyu BAI ; Shaohua DU ; Peng CHENG ; Ning KANG ; Zhen WANG ; Qiangkai HUANG ; Ziquan YANG
Chinese Journal of Tissue Engineering Research 2013;(36):6489-6494
BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.
3.Protective effect of HO-1 transfection against ethanol-induced osteoblast damage.
Jie LI ; Feng-Quan ZHANG ; Zhen-Ning DU ; Teng CAI ; Peng-Shan CAI ; Lei FAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(3):374-377
Heme oxygenase-1 (HO-1) plays important roles in anti-oxidant, anti-inflammatory and immunoregulative activities. The aim of this study was to observe if HO-1 transfection could inhibit the damage of osteoblasts induced by ethanol. HO-1 was transfected into osteoblasts via constructed plasmid. After exposure to ethanol for 24 h, cytoactivity and apoptosis of osteoblasts were measured by MTT assay and flow cytometry, respectively. Furthermore, the oxidative stress and inflammatory factors in osteoblasts were measured. Compared to positive control group, the cytoactivity of transfected osteoblasts was significantly increased, and the apoptosis rate was significantly decreased (P<0.05). At the same time, the levels of reactive oxygen species (ROS), methane dicarboxylic aldehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-1 (IL-1) were significantly decreased (P<0.05), and superoxide dismutase (SOD) level was increased (P<0.05) in the transfected osteoblasts as compared with positive controls. These results suggest that HO-1 plays a protective role in osteoblasts, and HO-1 transfection can effectively inhibit bone damage induced by ethanol.
Cells, Cultured
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Ethanol
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toxicity
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Gene Expression Regulation
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drug effects
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Genetic Vectors
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pharmacology
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Heme Oxygenase-1
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genetics
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metabolism
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Humans
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Osteoblasts
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cytology
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drug effects
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Oxidative Stress
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drug effects
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Transfection
4.Analysis of treatment and prognosis in 34 patients with anorectal malignant melanoma.
Zhen-Yu ZOU ; Hai-Liang LIU ; Pei-Ming SUN ; Ning NING ; Song-Yan LI ; Xiao-Hui DU
Chinese Journal of Gastrointestinal Surgery 2013;16(5):459-462
OBJECTIVETo explore the prognostic factors of anorectal malignant melanoma (ARMM).
METHODSMedical records and follow-up data of 34 patients with ARMM treated in the Chinese PLA General Hospital from March 1993 to November 2011 were analyzed retrospectively.
RESULTSThere were 26 abdominoperineal resections(APR) and 8 wide local excisions (WLE). Twenty patients underwent postoperative adjuvant therapy, including chemotherapy in 14 cases, radiotherapy in 2 cases, traditional Chinese medicine therapy in 4 cases and immunotherapy in 16 cases. Postoperative follow-up was carried out in all the patients and the mean follow-up period was 27 months. The 1-, 3- and 5-year overall survival rates were 76.3%, 39.6% and 20.6% respectively, while the 1-, 3- and 5-year disease-free survival rates were 60.6%, 30.8% and 12.8% respectively. APR and postoperative immunotherapy could significantly reduce the local recurrence rate. According to the Kaplan-Meier method, gross type of tumor, mural involvement, lymph metastasis, and clinical staging had significant effects on overall survival, while lymph metastasis and postoperative immunotherapy on disease-free survival. Cox proportional hazards model indicated that the clinical staging and postoperative immunotherapy were significant predictive factors.
CONCLUSIONSEarly diagnosis and correct choice of surgical method are the keys to the treatment. Postoperative immunotherapy can prolong disease-free survival.
Disease-Free Survival ; Humans ; Lymphatic Metastasis ; Melanoma ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Survival Rate
5.Robotic anterior resection of rectal cancer: technique and early outcome.
Xiao-hui DU ; Di SHEN ; Rong LI ; Song-yan LI ; Ning NING ; Yun-shan ZHAO ; Zhen-yu ZOU ; Na LIU
Chinese Medical Journal 2013;126(1):51-54
BACKGROUNDThe Da Vinci system is a newly developed device for colorectal surgery. With advanced stereoscopic vision, lack of tremor, and the ability to rotate the instruments surgeons find that robotic systems are ideal laparoscopic tools. Since conventional laparoscopic total mesorectal excision is a challenging procedure, we have sought to assess the utility of the Da Vinci robotic system in anterior resections for rectal cancer.
METHODSBetween November 2010 and December 2011, a total of 22 patients affected by rectal cancer were operated on with robotic technique, using the Da Vinci robot. Data regarding the outcome and pathology reports were prospectively collected in a dedicated database.
RESULTSThere were no conversions to open surgery and no postoperative mortality of any patient. Mean operative time was (220 ± 46) minutes (range, 152 - 286 minutes). The median number of lymph nodes harvested was (14.6 ± 6.5) (range, 8 - 32), and the circumferential margin was negative in all cases. The distal margin was (2.6 ± 1.2) cm (range, 1.0 - 5.5 cm). The mean length of hospital stay was (7.8 ± 2.6) days (range, 7.0 - 13.0 days). Macroscopic grading of the specimen was complete in 19 cases and nearly complete in three patients.
CONCLUSIONSRobotic anterior resection for rectal surgery is safe and feasible in experienced hands. Outcome and pathology findings are comparable with those observed in open and laparoscopy procedures. This technique may facilitate minimally invasive radical rectal surgery.
Aged ; Digestive System Surgical Procedures ; instrumentation ; methods ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Rectal Neoplasms ; surgery ; Rectum ; surgery ; Robotics ; methods ; Treatment Outcome
6.Association of sterol regulatory element binding protein 2 and insulin-like growth factor binding protein 3 genetic polymorphisms with avascular necrosis of the femoral head in the Chinese population.
Yang SONG ; Zhen-wu DU ; Qiu-ju LI ; Gui-zhen ZHANG ; Ling-ling WANG ; Ning WU ; Jin-cheng WANG ; Zhong-li GAO
Chinese Medical Journal 2012;125(22):4037-4043
BACKGROUNDSterol regulatory element binding protein (SREBP)-2 plays a key role in lipid homeostasis by stimulating gene expression of cholesterol biosynthetic pathways. The insulin-like growth factor binding protein (IGFBP) family regulates growth and metabolism, especially bone cell metabolism, and correlates with osteonecrosis. However, association of their gene polymorphisms with risk of avascular necrosis of the femoral head (ANFH) has rarely been reported. We determined whether SREBP-2 and IGFBP-3 gene polymorphisms were associated with increased ANFH risk in the Chinese population.
METHODSTwo single nucleotide polymorphisms of SREBP2 gene, rs2267439 and rs2267443, and one of IGFBP-3 gene, rs2453839, were selected and genotyped in 49 ANFH patients and 42 control individuals by direct sequencing assay.
RESULTSThe frequencies of rs2267439 TT and rs2267443 GA of SREBP2 and rs2453839 TT and CT of IGFBP-3 in the ANFH group showed increased and decreased tendencies (against normal control group), respectively. Interaction analysis of genes revealed that the frequency of carrying rs2267439 TT and rs2267443 GA genotypes of SREBF-2 in ANFH patients was significantly higher than in the control group (P < 0.05). Association analysis between polymorphisms and clinical phenotype demonstrated that the disease course in ANFH patients with the rs2453839 TT genotype of IGFBP-3 was significantly shorter than that of CT + CC carriers (P < 0.01). CT + CC genotype frequency in patients with stage III/IV bilateral hip lesions was significantly higher than in those with stage III/IV unilateral lesions and stage II/III bilateral lesions (P < 0.05 - 0.02).
CONCLUSIONSOur results suggested that interaction of SREBP-2 gene polymorphisms and the relationship between the polymorphisms and clinical phenotype of IGFBP-3 were closely related to increased ANFH risk in the Chinese population. The most significant finding was that the CT + CC genotype carriers of IGFBP-3 rs2453839 were highly associated with the development of ANFH.
Adult ; Asian Continental Ancestry Group ; genetics ; Female ; Femur Head Necrosis ; genetics ; Genetic Predisposition to Disease ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Sterol Regulatory Element Binding Protein 2 ; genetics
7.A novel de novo mutation of the Nipped-B-like gene in an isolated Chinese patient with Cornelia de Lange syndrome.
Wei-zhen XU ; Chun-yue CHEN ; Xiao-ling CHEN ; Yan ZHAO ; Wen-ting LIU ; Zhen-fang DU ; Xian-ning ZHANG
Chinese Medical Journal 2013;126(1):191-192
Child, Preschool
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De Lange Syndrome
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genetics
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Female
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Humans
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Mutation
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Proteins
;
genetics
8.Randomized controlled trial on manipulation for the treatment of cervical vertigo of high flow velocity type.
Hong-Gen DU ; Hui WEI ; Mei-Zhen HUANG ; Zhong JIANG ; Shu-Liang YE ; Hong-Quan SONG ; Ji-Wei YU ; Xi-Tao NING
China Journal of Orthopaedics and Traumatology 2010;23(3):212-215
OBJECTIVETo explore the effects of manipulation and traction combined with Nimodipine on the blood flow velocity of vertebrobasilar artery (VBA) in cervical vertigo of high flow velocity,and to evaluate clinical therapeutic effects between two methods.
METHODSFrom March 2008 to Feburary 2009,70 patients who were diagnosed as high flow velocity of cervical vertigo were randomly divided into treatment group (35 cases) and control group (35 cases). Among 70 patients, 32 were male and 38 were female. The age ranged from 21 to 45 years with an average of 37.6 years. The disease course ranged from one day to two years with an average of 12.6 days. Patients of the treatment group were treated with manipulation for total three weeks, three times once week. The patients in the control group were treated with traction (weight ranged from 5 to 6 kg, 20 minutes each time, once every other day) and Nimodipine for total three weeks (three times each day, and with a dose of 40 mg each time). After three weeks, the changes of flow velacity of VBI and score before and after treatment were observed using transcranil Doppler (TCD) and Evaluation Scale for Cervical Vertigo. After six weeks, the therapeutic effects were assessed.
RESULTSThe mean velocity in left vertebral artery (LVA), right vertebral artery (RVA) and basilar artery (BA) were obviously lower than those before treatment in two groups (P < 0.01). The LVA, RVA and BA of the treatment group was lower than those of control group after 3 weeks (P < 0.01). There was significant difference in vertigo score after treatment between the two groups. The improvement rate of double-sides sign in X-ray image and the therapeutic effects of treatment group was superior to that of control group (P < 0.01).
CONCLUSIONThe effect of manipulation on flow velocity of VBA is superior to that of traction combined with Nimodipine, and there are better therapeutic effects in treating cervical vertigo of high flow velocity in comparison with traction combined with Nimodipine. But there are more higher demands for manipulation's application.
Adult ; Arteries ; physiopathology ; Blood Flow Velocity ; Cervical Vertebrae ; blood supply ; Female ; Humans ; Male ; Middle Aged ; Musculoskeletal Manipulations ; Vertigo ; physiopathology ; therapy ; Young Adult
9.Clinical analysis of lumbar intervertebral disc herniation in adolescent with manipulation and traditional Chinese medicine.
Hong-gen DU ; Zhong JIANG ; Hui WEI ; Hong-quan SONG ; Shu-liang YE ; Mei-zhen HUANG ; Xi-tao NING
China Journal of Orthopaedics and Traumatology 2009;22(4):253-254
Adolescent
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Adult
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Drugs, Chinese Herbal
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therapeutic use
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Female
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Follow-Up Studies
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Humans
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Intervertebral Disc Displacement
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drug therapy
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physiopathology
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surgery
;
therapy
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Lumbar Vertebrae
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pathology
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Male
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Manipulation, Orthopedic
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Medicine, Chinese Traditional
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Recovery of Function
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Treatment Outcome