OBJECTIVETo investigate the effects of hydrogen sulfide (H2S) on H2O2-stimulated primary neonatal rat cardiomyocytes and related mechanism.

METHODSPrimary neonatal rat cardiomyocytes were treated with various concentrations of H2O2 (10, 100, 1000 µmol/L) for 24 h to establish the oxidative stress-induced cell injury model after 3 days' conventional culture. In addition, different concentrations of NaHS (1, 10, 100 µmol/L) were added to cardiomyocytes in the absence and presence of 100 µmol/L H2O2 for 24 h. The viability of cardiomyocytes was measured by MTT assay. The SOD vitality was measured by xanthine oxidase method and MDA content was determined by thiobarbituric acid colorimetric method. LDH activity was measured by chemical colorimetric method. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 (Rh123) staining and photofluorography. The level of reactive oxygen species (ROS) in cardiomyocytes was measured by DCFH-DA staining and photofluorography.

RESULTSCell viability and SOD vitality were significantly reduced while MDA content and LDH activity were significantly increased with increasing H2O2 concentrations. These effects could be partly reduced by cotreatment with H2O2 in a concentration-dependent manner (all P < 0.05). Compared with control group, the DCF fluorescence intensity significantly increased in the 100 µmol/L H2O2 group (P = 0.003), which could be attenuated by NaHS in a dose-dependent manner. Compared with control group, the MMP significantly decreased in the 100 µmol/L H2O2 group (P = 0.000), which could be partly reversed by cotreatment with NaHS in a dose-dependent manner. Moreover, H2O2 treatment also significantly reduced 100 µmol/L H2O2 induced apoptosis in a dose-dependent manner.

CONCLUSIONH2S protects primary neonatal rat cardiomyocytes against H2O2-induced oxidative stress injury through inhibition of H2O2 induced overproduction of ROS, dissipation of MMP and apoptosis.

Animals ; Cell Survival ; drug effects ; Cells, Cultured ; Hydrogen Peroxide ; pharmacology ; Hydrogen Sulfide ; pharmacology ; Malondialdehyde ; metabolism ; Membrane Potential, Mitochondrial ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Superoxide Dismutase ; metabolism

Purpose: The prognosis of nasopharyngeal carcinoma (NPC) patients with parotid lymph node (PLN) metastasis remains unclear. This study was performed to investigate the prognostic significance and optimal staging category of PLN metastasis and develop a nomogram for estimating individual risk. Materials and Methods: Clinical data of 7,084 non-metastatic NPC patients were retrospectively reviewed. Overall survival (OS) was the primary endpoint. A nomogram was established based on the Cox proportional hazards regression model. The accuracy and calibration ability of this nomogram was evaluated by C-index and calibration curves with bootstrap validation.ResultTotally, 164/7,084 NPC patients (2.3%) presented with PLNs. Multivariate analyses showed that PLN metastasis was a negative prognostic factor for OS, progression-free survival (PFS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRFS). Patients with PLN metastasis had a worse prognosis than N3 disease. Five independent prognostic factors were included in the nomogram, which showed a C-index of 0.743. The calibration curves for probability of 3- and 5-year OS indicated satisfactory agreement between nomogram-based prediction and actual observation. All results were confirmed in the validation cohort. Conclusion NPC patient with PLN metastasis had poorer survival outcome (OS, PFS, DMFS, and LRFS) than N3 disease. We developed a nomogram to provide individual prediction of OS for patients with PLN metastasis.

OBJECTIVESTo establish a clinical method for measuring the displacement of the distal radioulnar joint (DRUJ) precisely irrespective of ulnar variance, and to derive normal population translation references with palmar and dorsal stress.

METHODSThirty-seven normal distal forearms were scanned with computed tomography using an apparatus designed by Pirela-Cruz. Each extremity was scanned in two positions: maximal ulnar palmar and dorsal stress. The digital imaging and communications in medicine (DICOM) CT images were then imported into Mimics 10.0 for three-dimensional reconstruction. On the DRUJs 3D images, choose the most prominent point of the palmar and dorsal margins of the sigmoid notch and the excavate ulna fovea as the reference points A, B and C. A perpendicular line was then drawn from the point C to a line connecting points A and B with the intersection D. Calculate the ratio of AD/AB and DB/AB. Two observers measured all the DRUJs independently and one repeated the measurements one month later to determine the interobserver and intraobserver reliability.

RESULTSThe mean ratio values of palmar (AD/AB) and dorsal (DB/AB) translation were 0.39 ± 0.07 and 0.37 ± 0.07, and the normal references (x(-) ± 2 s) were from 0.25 to 0.50 and from 0.23 to 0.50, respectively. No significant differences were observed in terms of positions, genders and dominant hands. The intraclass correlation coefficient (ICC) values for interobserver and intraobserver reliability (DB/AB, AD/AB) were 0.84, 0.80, 0.93 and 0.92, respectively.

CONCLUSIONSThis new method could accurately measure the displacement of DRUJs with acceptable reliability, even with ulna positive or negative variance. Instability of DRUJ may be indicated when AD/AB is less than 0.25 or BD/AB is less than 0.23.

Adult ; Aged ; Female ; Humans ; Imaging, Three-Dimensional ; Joint Dislocations ; diagnostic imaging ; Joint Instability ; diagnostic imaging ; Male ; Middle Aged ; Radius ; diagnostic imaging ; Tomography, X-Ray Computed ; Ulna ; diagnostic imaging ; Wrist Joint ; diagnostic imaging ; Young Adult

OBJECTIVETo investigate the necessity of detecting on the expressive intensity and pattern of HBsAg and HBcAg in the livers of chronic hepatitis B.

METHODSHBsAg and HBcAg were detected in paraffin-embedded liver tissue by EnVision immunohistochemistry. Serum hepatitis B virus DNA (HBV DNA) was tested by real-time quantitative polymerase chain reaction. The degrees of hepatic inflammatory activity (grade) and fibrosis (stage) of liver biopsies were determined according to the standard of the Chinese program of prevention and treatment of viral hepatitis.

RESULTSThe expression of HBsAg was not correlated with the grade, the stage and the levels of serum HBV DNA (P > 0.05). Liver HBcAg expressive intensity was not correlated with the grade (r=0.02, P > 0.05), while negatively correlated with the stage (r=0.28, P < 0.01) and positively correlated with the serum HBV DNA levels (r=0.53, P < 0.01). Liver HBcAg expressive pattern was negatively correlated with the grade (r=-0.27, P < 0.01). The grade in cytoplasmic pattern group was higher than in nuclear pattern group and in mixed pattern group (P < 0.01), and that in mixed pattern group was higher in nuclear pattern group (P < 0.01). Liver HBcAg expressive pattern was negatively correlated with the stage (r=-0.23, P < 0.01). The stage in cytoplasmic pattern group was higher than in nuclear pattern group and in mixed pattern group (P < 0.05). Liver HBcAg expressive pattern was positively correlated with the levels of serum HBV DNA (r=0.22, P < 0.01).

CONCLUSIONDistinguishing the expressive intensity and pattern of HBsAg and HBcAg in the liver of chronic hepatitis B may not help understand the degree of hepatic lesion. The detection of HBcAg in liver tissue of CHB may be beneficial for the antiviral therapy.

Adult ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B Antigens ; biosynthesis ; Hepatitis B Core Antigens ; biosynthesis ; Hepatitis B Surface Antigens ; biosynthesis ; Hepatitis B virus ; genetics ; immunology ; physiology ; Hepatitis B, Chronic ; pathology ; virology ; Host-Pathogen Interactions ; Humans ; Immunohistochemistry ; Liver ; pathology ; virology ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Virus Replication ; Young Adult

PURPOSE: The purpose of this study was to subdivide M1 stage nasopharyngeal carcinoma (NPC) patients with bone-only metastases for prognosis prediction while identifying the treatment effect of locoregional radiotherapy (LRRT) and metastasis radiotherapy (MRT) among patients with different risk. MATERIALS AND METHODS: From November 2006 to October 2016, a total of 226 patients with bone-only metastasic NPC were retrospectively enrolled. All patients developed distant lesions before receiving treatment. All potential prognostic factors were considered and the correlation of the M1 subdivisions with overall survival (OS) was determined by Cox regression hazards model. Kaplan–Meier curves were used to appraise survival condition and log-rank testing was used to compare the differences. RESULTS: The median follow-up time was 33.9 months (range, 3 to 126 months). According to multivariate Cox proportional hazard analysis, the number of metastatic lesions and Epstein-Barr virus (EBV) DNA status after palliative chemotherapy (PCT) were independent prognostic factors for OS. Thus, we subdivided patients into three risk groups according to these two factors. Systemic chemotherapy combined with LRRT may benefit patients in low- and intermediate-risk groups but not in the high-risk group. Further aggressive MRT based on systemic chemotherapy showed no survival benefit in any risk group. CONCLUSION: The stratification of NPC patients with bone-only metastasis based on EBV DNA after PCT and the number of metastatic lesions provided promising prognostic value and could aid clinicians in person-specific treatment.
Diagnosis ; DNA ; Drug Therapy ; Follow-Up Studies ; Herpesvirus 4, Human ; Humans ; Neoplasm Metastasis ; Prognosis ; Proportional Hazards Models ; Radiotherapy ; Retrospective Studies

PURPOSE: The purpose of this study was to investigate the survival trends and patterns of failure in patients with stage II nasopharyngeal carcinoma (NPC) treated with radiotherapy (RT) and chemotherapy over the last 20 years. MATERIALS AND METHODS: Thirty-eight hundred and eight patients diagnosed with stage II NPC between January 1990 and December 2012 were involved in this retrospective cohort study. All patients were treated with RT. According to the main imaging techniques and RT technology, we categorized these patients into four calendar periods: 1990-1996, 1997-2002, 2003-2007, and 2008-2012. Overall survival (OS), progression-free survival (PFS), locoregional relapse-free survival (LRFS), and distant metastasis–free survival (DMFS) were served as the clinical outcome. RESULTS: After a median follow-up period of 84.7 months, we observed increasing trends in survival and disease control. The 3- and 5-year OS rates increased from 87.1% and 78.7% in the first calendar period to 97.4% and 94.5% in the last calendar period, respectively (p<0.001). Additionally, significant increasing trends could be seen in the PFS and LRFS during the four calendar periods. In the subgroup analysis, the LRFS in patients older than 50 years at diagnosis showed greater improvement than younger patients. However, the rate of distant metastasis was stable and relatively low, as the 5-year DMFS ranged from 90.5% to 94.7% among the four calendar periods. CONCLUSION: The survival rates in patients with stage II NPC showed increasing trends from 1990 to 2012. The advance of RT provided excellent locoregional control and enhanced OS.
Cohort Studies ; Diagnosis ; Disease-Free Survival ; Drug Therapy ; Follow-Up Studies ; Humans ; Neoplasm Metastasis ; Prognosis ; Radiotherapy ; Retrospective Studies ; Survival Rate

In this study, we investigated the effects of Panax notoginseng saponins (PNS) on pulmonary vascular remodeling and ADAM10/Notch3 pathway in pulmonary arterial hypertension (PAH). PAH rat model was established, and male Sprague Dawley (SD) rats were randomly divided into control group, monocrotaline (MCT) group and MCT+PNS group, with 10 rats in each group. Rats in the control group were intraperitoneally injected with equal volume of normal saline. Rats in the MCT group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with the same volume of normal saline every day. Rats in the MCT+PNS group was injected intraperitoneally with 60 mg/kg MCT on the first day, and then with 50 mg/kg PNS every day. The modeling time of each group lasted for 21 days. After the model was established, the mean pulmonary artery pressure (mPAP) was measured by right heart catheterization technique, the right ventricular hypertrophy index (RVHI) was calculated, the microscopic morphology and changes of pulmonary vascular wall were observed by HE and Masson staining, and the expressions of ADAM10, Notch3, Hes-1, P27, PCNA, Caspase-3 proteins and mRNA in pulmonary vascular tissue of rats were detected by Western blot and qPCR. The expression and localization of Notch3 and α-SMA were detected by immunofluorescence staining. The protein expression of ADAM10 was detected by immunohistochemical staining. The results showed that compared with the control group, mPAP, RVHI, pulmonary vessels and collagen fibers in the MCT group were significantly increased, the expressions of ADAM10, Notch3, Hes-1, and PCNA protein and mRNA were significantly increased, while the expressions of P27 and Caspase-3 protein and mRNA were decreased significantly. Compared with the MCT group, mPAP and RVHI were significantly decreased, pulmonary vessels were significantly improved and collagen fibers were significantly reduced, the expressions of protein and mRNA of ADAM10, Notch3, Hes-1, and PCNA were decreased in MCT+PNS group, but the expressions of protein and mRNA of P27 and Caspase-3 were increased slightly. The results of immunofluorescence showed that Notch3 and α-SMA staining could overlap, which proved that Notch3 was expressed in smooth muscle cells. The expression of Notch3 in the MCT group was increased significantly compared with that in the control group, while PNS intervention decreased the expression of Notch3. Immunohistochemical staining showed that compared with the control group, the amount of ADAM10 in the MCT group was increased significantly, and the expression of ADAM10 in the MCT+PNS group was decreased compared with the MCT group. These results indicate that PNS can improve the PAH induced by MCT in rats by inhibiting ADAM10/Notch3 signaling pathway.
Animals ; Male ; Rats ; Caspase 3/metabolism* ; Collagen ; Disease Models, Animal ; Hypertension, Pulmonary/drug therapy* ; Monocrotaline/adverse effects* ; Panax notoginseng/chemistry* ; Proliferating Cell Nuclear Antigen/pharmacology* ; Pulmonary Arterial Hypertension ; Pulmonary Artery/metabolism* ; Rats, Sprague-Dawley ; Receptor, Notch3/genetics* ; RNA, Messenger ; Saline Solution ; Signal Transduction ; Saponins/pharmacology*

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

OBJECTIVETo investigate the factors related to clinical pregnancy outcomes of in vitro fertilization-embryo transfer (IVF-ET) in women with secondary infertility.

METHODSThe clinical, laboratory and follow-up data of 1129 cycles in 1099 patients with secondary infertility undergoing IVF-ET in Women's Hospital, Zhejiang University School of Medicine between July 2012 to July 2014 were retrospectively reviewed. The factors related to pregnancy outcomes were analyzed by univariate and logistic regression methods. The clinical pregnancy rates in women with different age and different number of embryos transferred were compared. The clinical outcomes of stimulation with gonadotropin releasing hormone (GnRH) agonist long protocol, GnRH agonist short protocol and GnH antagonist protocol were evaluated in secondary infertile patients aged ≥ 40 years.

RESULTSAmong 1129 cycles, 376 cases (33.30%) had clinical pregnancy and 753 cases (66.70%) had no clinical pregnancy. There were significant differences in age, body mass index, basal follicle-stimulating hormone level, antral follicle number,paternal age and number of embryos transferred between pregnancy and no pregnancy groups (P<0.05); while only maternal age (OR=0.900, 95% CI: 0.873~0.928, P<0.001) and the number of embryos transferred (OR=2.248, 95% CI: 1.906~2.652, P<0.001) were the independent factors affecting the clinical pregnancy outcome of IVF-ET. There was no significant difference in clinical pregnancy rate between women aged 30~40 years with two embryos transferred and those aged<30 years with two or three embryos transferred(P>0.05). There were no significances in clinical pregnancy rate among women aged ≥ 40 years using GnRH agonist long protocol,GnRH agonist short protocol and GnRH antagonist protocol for stimulation (P>0.05).

CONCLUSIONMaternal age and number of embryos transferred have independent effect on IVF-ET clinical pregnancy outcome of secondary infertile women. We suggest that no more than two embryos should be transferred for women in their thirties to minimize the risk of multiple pregnancy while still having an acceptable pregnancy rate. The pregnancy rate of patients over 40 years decreases significantly, and there is no difference in pregnancy rate by using GnRH agonist long protocol, GnRH agonist short protocol or GnRH antagonist protocol.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; Gonadotropin-Releasing Hormone ; agonists ; Gonadotropins ; Hormone Antagonists ; therapeutic use ; Humans ; Infertility, Female ; Maternal Age ; Ovarian Follicle ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies