Objective: To explore the effects of fosinopril on oxidative stress and vascular function in experimental rats with spontaneous hypertension. Methods: The rats were divided into 3 groups: Control group, with normal healthy rats (n=15), Spontaneous hypertension (SH) group (n=15), SH rats received intragastric administration of normal saline and Treatment group (n=15), SH rats received intragastric administration of fosinopril 10mg/(kg?d). All animals were treated for 7 weeks. Caudal artery systolic blood pressure (SBP) was measured at each week. blood levels of superoxide dismutase (SOD), reactive oxygen species (ROS), malonaldehyde (MDA) and NO2-/NO3- were determined in different groups respectively after 7 weeks. Moreover, thoracic aorta was taken to examine its diastolic reactive rate by acetylcholine (Ach)/sodium nitroprusside (SNP) induction. Results: From the 1st week until the end of experiment, compared with SH group, Treatment group had decreased SBP,P<0.05. With 7 weeks treatment, compared with Control group, SH group had decreased SOD activity, while increased protein levels of MDA and ROS, allP<0.05; compared with SH group, Treatment group showed elevated SOD activity (P=0.010), while reduced protein levels of MDA (P=0.021) and ROS (P=0.009). Compared with Control group, SH group had the lower content of NO2-/NO3-(P<0.001); both SH group and Treatment group had decreased diastolic rates by Ach/SNP induction,P<0.05. Compared with SH group, Treatment group presented the higher content of NO2-/NO3- and higher diastolic rate by Ach induction, allP<0.001. Conclusion: Fosinopril could improve vascular diastolic function via anti-oxidative stress in experimental SH rats, which might be one of its anti-hypertensive mechanisms.

BACKGROUND: It is one of hot topics for the application of neurotrophic factors including neurenergen-3 to promote peripheral neural regeneration nowadays; however, clinical application is restricted to safety and effective administration route. Gene transfection brings a novel thinking and pathway for neurotrophic factors used in clinic.OBJECTIVE: To observe the expression of neural stem cells modified by neurenergen-3 gene after transfection.DESIGN: Completely randomized study.SETTING: Department of Traumatic Orthopaedics and Hand Surgery, the First Affiliated Hospital, Guangxi Medical University;Department of Hand Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Three healthy SD rats of four months old and either gender were selected from Animal Center, Tongji Medical College, Huazhong University of Science and Technology. Recombinant adenoviral expressing vector for transfection of neural stem cells was constructed in Laboratory of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, and the concentration was 0.15 g/L.METHODS: The experiment was carried out in the Orthopaedic Laboratory and Central Laboratory, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology from December 2002 to March 2004. Recombinant plasmid pAD-neurenergen-3 containing with green fluorescent protein (GFP) gene was transfectd into primarily cultured neural stem cells by using cationic tiposome interventional method. At 72 hours after transfection, fluorescent inverted microscope was used to observe GFP expression in neural stem cells, and transfection efficiency was measured simultaneously. Expression and transcription of neurenergen-3 gene in neural stem cells were detected at 72 hours, 1 and 5 weeks after transfection by using immunocytochemical stain and reverse transcription polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of neurenergen-3 gene in transfected neural stem cells.RESULTS: ① Neural stem cells transfected by recombinant plasmid pAD-neurenergen-3: GFP expressed on partial neural stem cells at 72 hours after transfection, and the transfection efficiency was 40%. Five weeks later, GFP expression was still observed. ②Transcription and expression of neurenergen-3 gene in neural stem cells: Transcription of neurenergen-3 mRNA was observed in neural stem cells at 72 hours, 1 and 5 weeks after transfection, and expression of neurenergen-3 mRNA was still observed in 5 weeks after trans fection.CONCLUSION: As the carrier of cationic liposome, neurenergen-3 gene can effectively transfect, culture and long-term express neural stem cells via the introduction of adenoviruses.

ObjectiveTo investigate the expression and activity of histone deacetylase (HDAC) in prostate cancer.Methodshe pathological samples of 37 cases of PCa were collected. The mean age of the patients was 73 (53 - 88 ) years, the preoperative t-PSA was 81.69 ( 3.13 - 2000 ) ng/ml, Gleason score: 13 cases were ≤7, 24 cases were ＞7. Twenty-seven cases of BPH were set as controls. The mean age of the BPH patients was 69 (52 - 84) years, the preoperative t-PSA was 10.93 ( 1.11 - 55.07 ) ng/ml.Western blotting and colorimetric Assay kits were used to determine the HDAC expression and activity. The difference of HDAC activity in benign prostatic hyperplasia and prostate cancer was statistically analyzed.The correlation of the HDAC expression level and values of PSA and Gleason score was also assessed.ResultsHDACs were over-expressed in most cases of prostate cancer, the expression rates were HDAC1 :57％, HDAC2: 68％, HDAC3: 84％ and HDAC4: 73％, respectively. The HDAC activity (P ＜0.05)was significantly different between the prostate cancer and benign prostatic hyperplasia groups. The expression level of HDAC did not correlate with the values of PSA and Gleason score.ConclusionsHDAC was highly expressed and strongly active in prostate cancer. The results suggest that HDAC might be a potential target for the management of prostate cancer patients.

Objective To establish rat models of polycystic ovary syndrome(PCOS) induced by different methods,to assess the serum levels of several related hormones,to examine the morphological changes in the ovaries,and to discuss their significance.Methods Letrozol,sodium prasterone sulfate,or sodium prasterone sulfate combined with human chorionic gonadotropin were used to establish rat models of PCOS.Radioimmunoassay was used to measure the serum levels of hteinizing hormone(LH),follicle-stimulating hormone(FSH),estrogen(E_2),progesterone(P),testosterone(T),prolactin(PRL),and insulin(INS).HE staining was used to examine the morphological changes of the ovaries.Results Comparing with the normal group A,the serum FSH was increased and the serum progesterone was reduced in the group B,with a statistically significant difference(P<0.05).The serum testosterone was significantly higher in the group B than in the group A(P<0.01).The levels of serum sex hormones and insulin were not significantly different in the group D and C(P>0.05).In comparison with the group C,the levels of serum testosterone and LH/FSH ratio was significantly increased in the group E.(P<0.05).Comparing with the group D,the serum levels of progesterone and testosterone were significantly increased in the group E(P<0.05).The ovaries in the rats of groups A and C showed almost a normal histyology,with mature follicles and dominant follicles.Polycystic changes were observed only in the ovaries of groups B,D and E.Conclusion At the aspect of affecting the level of sex hormones in serum and changing the ovarian morphology.adopting letrozol tablets or sodium prasterone sulfate combined with HCG to induce rat PCO model is more close to clinic manifestations and meets the criteria of PCO animals.In the rat PCOS models induced with letrozol or with sodium prasterone sulfate combined with HCG,either the serum levels of sex hormones and ovarian histology are quite similar to those of human clinical appearance,and may well meet the modeling requirements for future experimental studies of polycystic ovary syndrome.

Objective To investigate the effects of Jianpi Huatan Fang on lipid metabolism and expression levels of FOXO1 and PDK4 in rats with polycystic ovary syndrome(PCOS)and insulin resistance(IR).Methods Forty female rats were divided randomly into a blank control group(n=10)and a model group(n=30).A PCOS-IR rat model was established using a high-fat diet(8 weeks)combined with letrozole(added at weeks 4～8).Thirty successfully modeled rats were then divided randomly into a model control group,a Jianpi Huatan Fang group(11.07 g/kg),and a metformin group(0.2 g/kg)(n=10 rats per group).Drug intervention was given for 4 weeks.The general status,Traditional Chinese medicine syndrome score,and weight changes were observed in each group.Histopathological changes in the ovary were detected by hematoxylin and eosin staining.Fasting blood glucose and blood lipids were measured using a blood biochemical analyzer.Levels of sex hormones including follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),estradiol(E2),and insulin were determined by enzyme linked immunosorbent assay and changes in the expression of FOXO1 and PDK4 in the ovaries were determined by Western Blot and real-time fluorescence quantitative polymerase chain reaction.Results Rats in the model control group exhibited symptoms of spleen deficiency and phlegm-dampness and showed significant weight gain,sex hormone disorders,polycystic ovarian changes,and dysregulation of glucose and lipid metabolism,compared with rats in the blank control group(P＜0.01).In addition,weight increase was slower in rats in the Jianpi Huatan Fang group and the metformin group compared with that in the model control group,and follicle development was improved.Serum LH,T,LH/FSH,fasting blood glucose,insulin,homeostatic model assessment for IR,cholesterol,triglycerides,and low-density lipoprotein cholesterol decreased,estradiol and FSH increased,and ovarian FOXO1 and PDK4 mRNA and protein expression levels were down regulated in the Jianpi Huatan Fang group compared with the findings in the model control group(P＜0.05,P＜0.01).Serum high-density lipoprotein cholesterol content was also increased,but the difference was not significant(P＞0.05).Conclusions Jianpi Huatan Fang can effectively regulate sex hormone secretion,improve ovarian reproductive function,and regulate glucose and lipid metabolism in obese PCOS-IR rats,possibly via inhibiting the FOXO1/PDK4 pathway.

Objective To investigate the safety and feasibility of en-bloc resection of a primary sacral chordona based on a 3-dimensional printing model.Methods 31 patients with primary sacral chordoma underwent en-bloc resection via a onestage posterior approach or combined anterior and posterior approaches in our oncology department from January 2013 to December 2014.They comprised 21 males and 10 females of mean age (49.2±12.5) years (range,26-67 years).Preoperative 3-D printing models were created by 3D printing technology,it included tumor tissue,the around vascular and nerves involved in sacral chordoma.The sacral chordomas were en-bloc resection with decompression and internal fixation.Results With the mean (29.0±6.8)months follow-up (range from 19 to 41),all patients underwent en bloc excision via 26 cases with posterior approach,5 cases combined posterior and anterior approaches in one stage.The mean operative time and estimated blood loss were (275.0±58.1) min and (3 250.0±1 304.4) ml,respectively.The visual analogue scale (VAS) score was (5.6±1.9) in average (range from 3 to 9) at preoperation,and (2.0±1.5) at post-operation,which was significantly lower than that of preoperation,and the pain was relief obviously.There were 13 cases in grade C,11 cases in grade D,7 cases in grade E of American Spinal Injury Association (ASIA) grade neurological function before surgery,compared with the pre-operation,there were 5 cases in grade C,6 cases in grade D,20 cases in grade E of post-operation,which was significantly improved.MSTS (Musculoskeletal Tumor Society) 93 score was 6-29 points (20.0％-96.7％) at the follow-up 3 months after surgery,with the average of (19.8 ± 5.8) points,which excellent in 8 cases,good in 14 cases,general in 5 cases,poor in 4 cases.Two cases of dysporia for the reasons of resecting on one side of the S1,2 nerve roots involved by the sacral chordoma,after sacrificing the nerve root of complete tumor resection,the urine left dysfunctional,while the pain of other 29 patients were thoroughly relief after surgery.The ones were relieved with the disturbance of sensation of the perineum before the operation.2 cases were recovery of leakage of cerebrospinal by the drainage of lumbar cistern with normal temperature.One hypostatic pneumonia patient was cured by anti-inflammatory.One with the urinary infection got better by the effective bladder irrigation,which had diabetics mellitus with the bladder stoma before.1 case of skin necrosis due to vascular thrombosis before operation,recevied flap translocation half month after surgery,got recovery 3 months later.Only one underwent tumor resection for the recurrence at 15 months follow-up.Conclusion It is feasible and safe to perform en bloc resection of primary sacral chordoma.This is the most effective means of managing method of the marginal resection of the tumor.Preoperative 3-D printing modeling enables better anatomical understanding of the relationship between the tumor,and can avoid vascular and nerves tissue injury,which can also assist in planning the surgical procedure,and be worth recommendation.

OBJECTIVE: To investigate the effect of inhibitor of growth protein-2 (Ing2) silencing on angiotensin Ⅱ (AngⅡ)-induced cardiac remodeling in mice and explore the underlying mechanism. METHODS: An adenoviral vector carrying Ing2 shRNA or empty adenoviral vector was injected into the tail vein of mice, followed 48 h later by infusion of 1000 ng · kg^-1 · min^-1 Ang Ⅱ or saline using a mini-osmotic pump for 42 consecutive days. Transthoracic echocardiography was used to assess cardiac geometry and function and the level of cardiac hypertrophy in the mice. Masson and WGA staining were used to detect myocardial fibrosis and cross-sectional area of cardiomyocytes, and myocardial cell apoptosis was detected with TUNEL assay. Western blotting was performed to detect myocardial expressions of cleaved caspase 3, ING2, collagen Ⅰ, Ac-p53(Lys382) and p-p53 (Ser15); Ing2 mRNA expression was detected using real-time PCR. Mitochondrial biogenesis, as measured by mitochondrial ROS content, ATP content, citrate synthase activity and calcium storage, was determined using commercial assay kits. RESULTS: The expression levels of Ing2 mRNA and protein were significantly higher in the mice with chronic Ang Ⅱ infusion than in saline-infused mice. Chronic infusion of AngⅡ significantly increased the left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD) and reduced left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) in the mice. Ing2 silencing obviously alleviated AngⅡ-induced cardiac function decline, as shown by decreased LVEDD and LVESD and increased LVEF and LVFS, improved myocardial mitochondrial damage and myocardial hypertrophy and fibrosis, and inhibited cardiomyocyte apoptosis. Chronic AngⅡ infusion significantly increased myocardial expression levels of Ac-p53(Lys382) and p-p53(Ser15) in the mice, and Ing2 silencing prior to AngⅡ infusion lessened AngⅡ- induced increase of Ac-p53(Lys382) without affecting p53 (ser15) expression. CONCLUSION Ing2 silencing can inhibit AngⅡ-induced cardiac remodeling and dysfunction in mice by reducing p53 acetylation.
Animals ; Mice ; Angiotensin II ; Tumor Suppressor Protein p53 ; Acetylation ; Stroke Volume ; Ventricular Remodeling ; Ventricular Function, Left ; Myocytes, Cardiac

Objective:To investigate the clinical characteristics of pediatric methicillin-resistant Staphylococcus aureus (MRSA) infection and the antibiotic sensitivity of the isolates. Methods:The clinical data of children with MRSA infection and antibiotic sensitivity of the isolates from 11 children′s hospitals in Infectious Diseases Surveillance of Paediatrics (ISPED) group of China between January 1, 2018 and December 31, 2018 were collected retrospectively. The children′s general condition, high-risk factors, antimicrobial therapy and prognosis, differences in clinical disease and laboratory test results between different age groups, and differences of antibiotic sensitivity between community-acquired (CA)-MRSA and hospital-acquired (HA)-MRSA were analyzed. The t test and Wilcoxon rank sum test were used for statistical analysis of the quantitative data and Chi-square test were used for comparison of rates. Results:Among the 452 patients, 264 were males and 188 were females, aged from 2 days to 17 years. There were 233 cases (51.5%) in the ≤1 year old group, 79 cases (17.5%) in the>1-3 years old group, 29 cases (6.4%) in the >3-5 years old group, 65 cases (14.4%) in the >5-10 years old group, and 46 cases (10.2%) in the>10 years old group. The main distributions of onset seasons were 55 cases (12.2%) in December, 47 cases (10.4%) in February, 46 cases (10.2%) in November, 45 cases (10.0%) in January, 40 cases (8.8%) in March. There were 335 cases (74.1%) CA-MRSA and 117 (25.9%) cases HA-MRSA. Among all cases, 174 cases (38.5%) had basic diseases or long-term use of hormone and immunosuppressive drugs. During the period of hospitalization, 209 cases (46.2%) received medical interventions. There were 182 patients (40.3%) had used antibiotics (β-lactams, glycopeptides, macrolides, carbapenems, oxazolones, sulfonamides etc) 3 months before admission. The most common clinical disease was pneumonia (203 cases), followed by skin soft-tissue infection (133 cases), sepsis (92 cases), deep tissue abscess (42 cases), osteomyelitis (40 cases), and septic arthritis (26 cases), suppurative meningitis (10 cases). The proportion of pneumonia in the ≤1 year old group was higher than the >1-3 years old group,>3-5 years old group,>5-10 years old group,>10 years old group (57.5% (134/233) vs. 30.4% (24/79), 31.0% (9/29), 38.5% (25/65), 23.9% (11/46), χ 2=17.374, 7.293, 7.410, 17.373, all P<0.01) The proportion of skin and soft tissue infections caused by CA-MRSA infection was higher than HA-MRSA (33.4% (112/335) vs. 17.9% (21/117), χ 2=10.010, P=0.002), and the proportion of pneumonia caused by HA-MRSA infection was higher than CA-MRSA (53.0% (62/117) vs. 42.1% (141/335), χ 2=4.166, P=0.041). The first white blood cell count of the ≤1 year old group was higher than that children > 1 year old ((15±8)×10 9/L vs. (13±7)×10 9/L, t=2.697, P=0.007), while the C-reactive protein of the ≤1 year old group was lower than the 1-3 years old group,>5-10 years old group,>10 years old group (8.00 (0.04-194.00) vs.17.00 (0.50-316.00), 15.20 (0.23-312.00), 21.79(0.13-219.00) mg/L, Z=3.207, 2.044, 2.513, all P<0.05), there were no significant differences in procalcitonin (PCT) between different age groups (all P>0.05). After the treatment, 131 cases were cured, 278 cases were improved, 21 cases were not cured, 12 cases died, and 10 cases were abandoned. The 452 MRSA isolates were all sensitive to vancomycin (100.0%), linezolid (100.0%), 100.0% resistant to penicillin, highly resistant to erythromycin (85.0%, 375/441), clindamycin (67.7%, 294/434), less resistant to sulfonamides (5.9%, 23/391), levofloxacin (4.5%, 19/423), gentamicin (3.2%, 14/438), rifampicin (1.8%, 8/440), minocycline (1.1%, 1/91). The antimicrobial resistance rates were not significantly different between the CA-MRSA and HA-MRSA groups (all P>0.05). Conclusions:The infection of MRSA is mainly found in infants under 3 years old. The prevalent seasons are winter and spring, and MRSA is mainly acquired in the community. The main clinical diseases are pneumonia, skin soft-tissue infection and sepsis. No MRSA isolate is resistant to vancomycin, linezolid. MRSA isolates are generally sensitive to sulfonamides, levofloxacin, gentamicin, rifampicin, minocycline, and were highly resistant to erythromycin and clindamycin. To achieve better prognosis. clinicians should initiate anti-infective treatment for children with MRSA infection according to the clinical characteristics of patients and drug sensitivity of the isolates timely and effectively.