The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

Objective:To investigate the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in the lung tissue of rats with acute diquat (DQ) poisoning and the distribution of diquat in lungs.Methods:Fifty-four fasted male Wistar rats were randomized into control group ( n=6) and exposure group ( n=48) . According to the time point, the exposure group was divided into 2 h, 4 h, 12 h, 1 d, 3 d, 7 d, 11 d and 14 d groups with 6 rats in each group. Exposure groups were administered 11.55 mg/kg DQ (1 ml/100 g BW) by single-dose of intragastric administration, while the control group rats were given normal saline. The histopathological changes of lung tissue of rats in each group were observed. The expression of nrf2 in lung tissue was detected by immunohistochemistry, and the diquat concentration in lungs was determined by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) . Results:In the exposure group, DQ was detected in lungs on 2 hours after poisoning. The concentration of DQ in lung tissue decreased gradually over time, and there was no accumulation in lung tissue. The histopathological changes of lung tissue were not obvious in the early stage of poisoning. The injury was the most serious on the 3rd day, a large number of inflammatory cells could be seen in alveolar cavity and lung stroma, and the pathological injury of lung tissue began to be alleviated on the 7th day. The results of immunohistochemistry showed that Nrf2 was mainly expressed in the nucleus of pulmonery cells. The expression of Nrf2 in the exposure group was significantly higher than the control group. The expression of Nrf2 increased significantly at the 12th hour ( P<0.05) , reached the peak on the 3rd day ( P<0.05) . There was no difference between the control group and the 14th day ( P>0.05) . Conclusion:There was no accumulation of DQ in the lung tissue for a long time, and there was a hysteresis in lung injury induced by redox reaction of DQ. Nrf2 was highly expressed in the lung tissue of rats with acute DQ poisoning, which was correlated with histopathology injury of lung tissue, suggesting that Nrf2 plays an important role in antagonizing acute lung injury induced by DQ.

Objective:To investigate the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in the lung tissue of rats with acute diquat (DQ) poisoning and the distribution of diquat in lungs.Methods:Fifty-four fasted male Wistar rats were randomized into control group ( n=6) and exposure group ( n=48) . According to the time point, the exposure group was divided into 2 h, 4 h, 12 h, 1 d, 3 d, 7 d, 11 d and 14 d groups with 6 rats in each group. Exposure groups were administered 11.55 mg/kg DQ (1 ml/100 g BW) by single-dose of intragastric administration, while the control group rats were given normal saline. The histopathological changes of lung tissue of rats in each group were observed. The expression of nrf2 in lung tissue was detected by immunohistochemistry, and the diquat concentration in lungs was determined by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS) . Results:In the exposure group, DQ was detected in lungs on 2 hours after poisoning. The concentration of DQ in lung tissue decreased gradually over time, and there was no accumulation in lung tissue. The histopathological changes of lung tissue were not obvious in the early stage of poisoning. The injury was the most serious on the 3rd day, a large number of inflammatory cells could be seen in alveolar cavity and lung stroma, and the pathological injury of lung tissue began to be alleviated on the 7th day. The results of immunohistochemistry showed that Nrf2 was mainly expressed in the nucleus of pulmonery cells. The expression of Nrf2 in the exposure group was significantly higher than the control group. The expression of Nrf2 increased significantly at the 12th hour ( P<0.05) , reached the peak on the 3rd day ( P<0.05) . There was no difference between the control group and the 14th day ( P>0.05) . Conclusion:There was no accumulation of DQ in the lung tissue for a long time, and there was a hysteresis in lung injury induced by redox reaction of DQ. Nrf2 was highly expressed in the lung tissue of rats with acute DQ poisoning, which was correlated with histopathology injury of lung tissue, suggesting that Nrf2 plays an important role in antagonizing acute lung injury induced by DQ.

Objective Aims to reduce the hidden risks of laboratory biosafety,understand the status of biosafety laboratory record filing situations in Shenzhen,and also to provide scientific basis for further standardizing the management of biosafety laboratory in Shenzhen.Methods In 2017,75 laboratories in Shenzhen completed record filing were surveyed,method ologies adopted including application materials review,phone call consultation and communication,carrying out corrective ac tions based on feedback peer review suggestions and finally complete the record filing.Results The first/second level laborato ry of biosafety in shenzhen is mainly public medical institutions,followed by private hospitals.In 2017,the first three recordfiling LABS were Futian district,nanshan district and longgang district.According to the data analysis,lack of the second category of pathogenic microorganism laboratory activity project risk assessment report,and laboratory layout diagram function partition is not clear were two of the more prominent problems in the software and hardware of laboratory management respectively.Conclusions Basically,the overall record filing of Shenzhen biosafety laboratory is good,however,more measurements should be developed to deal with identified problems to further strengthen the standardized management of laboratory biosafety.

A new species of Alternaria causing leaf spots on the rubber tree (Hevea brasiliensis) in Yunnan, China, was isolated, examined, and illustrated. Morphologically, it belongs to the section Porri of Alternaria, which produces relatively large conidia and a simple or branched, filamentous long beak. It is, however, characterized by conidiophores gradually enlarging near the apex into a clavate conidiogenous cell and long ellipsoid to obclavate, smooth-walled conidia with a long filamentous beak. Molecular phylogenetic analyses based on ITS rDNA, GAPDH, and TEF1-alpha sequences demonstrate that the phytopathogen falls in the clade of the section Porri, being most closely related to A. sidae, A. sennae, A. deseriticola, A. cyamopsidis, A. rostellata, A. nitrimali, A. crassa, and A. thunbergiae.
Acanthaceae ; Accidental Falls ; Alternaria ; Animals ; Ascomycota ; Beak ; China ; Classification ; DNA, Ribosomal ; Hevea ; Rubber ; Spores, Fungal

AIM:To observe the expression of microRNA-126-5p during myocardial injury and its role in myo-cardial cell injury induced by adriamycin(also called doxorubicin, DOX).METHODS: The BALB/c mouse model of DOX-induced acute and chronic myocardial injury was established via intraperitoneal injection of DOX.HE staining was applied to observe the morphological changes of myocardial tissues.Lactate dehydrogenase(LDH)in serum was detected and PowerLab system was used to detect the influence of DOX on the changes of ±dp/dtmax.The expression of microRNA-126-5p in injured myocardial tissues and the H 9c2 cells exposed to DOX was detected by real-time PCR.Gain-and loss-of-function experiments were conducted to detect the role of microRNA-126-5p in H9c2 cells treated with DOX on LDH release and caspase-3 activation.RESULTS:In acute and chronic DOX myocardial damage models in mice,HE staining showed disarranged myocardial fibers, dissolved myofibril and inflammatory cell infiltration.Higher serum LDH level and lower ±dp/dtmaxin DOX-treated mice than those in normal mice were found.Compared with the normal mice, the expression level of microRNA-126-5p was significant increased in the myocardium with DOX-induced injury.Similarly,the expression level of microRNA-126-5p was significant increased in the H9c2 cells treated with DOX.In addition, over-expression of microRNA-126-5p decreased cell viability and promoted apoptosis,while microRNA-126-5p ablation promoted the viability and inhibited the apoptosis of H9c2 cells.CONCLUSION:The microRNA-126-5p expression is up-regulated in myocar-dial injury induced by DOX,and microRNA-126-5p inhibits cell viability and promotes apoptosis induced by DOX.

OBJECTIVETo investigate the presence of leukemia stem cells (LSC) in acute myeloid leukemia (AML) and find out the relative position of leukemia cells in the figures of flow cytometry, and to analyze the relationship between minimal residual diseases (MRD) and the level of LSC, so as to explore the correlation of LSC changes with the curative effect and the prognosis during chemical therapy.

METHODSA total of 85 samples were collected from 50 AML (except M3) patients, including 50 samples from the newly diagnosed patients, 7 samples of AML patients with non-remission and 28 samples of AML patients with complete remission. All samples were used for detection of LSC from immune phenotype of CD34/CD38/CD123 by flow cytometry. The detection of immune phenotypic of leukemia cells was performed in the newly diagnosed patients. The detection of leukemia- associated immune phenotypes (LAIP) was implemented in the non-newly diagnosed patients.

RESULTSThe LSC was identified in the CD34/ CD38/ CD123 in AML and consistent with the relative position of the leukemia cell in flow cytometry figures. Statistical analysis showed significant difference in LSC content between the newly diagnosed AML group and the post-chemotherapy complete remission group(P<0.01),but did not between the newly diagnosed AML group and the post-chemotherapy non-remission group(P>0.05).There was significant positive correlation between the LSC content and MRD level in 28 AML patients with complete remission (r=0.680,P<0.01).

CONCLUSIONLSC exist in AML and the relative position are consistent with the leukemia cells in flow cytometry figures, the size characteristics and weak expression of CD45 are also similar to leukemia cells. The proportion of LSC decreases after chemotherapy. Detecting and tracking the LSC changes in bone marrow and combination with detecting minimal resident disease(MRD) may contribute to evaluate the theraputic efficacy and prognosis of leukemia patients.

Flow Cytometry ; Humans ; Interleukin-3 Receptor alpha Subunit ; Leukemia, Myeloid, Acute ; Neoplasm, Residual ; Neoplastic Stem Cells ; Prognosis

Objective:To compare the long-term prognosis of percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) in patients with ostial/shaft lesions of unprotected left main coronary artery (ULMCA).Methods:A total of 259 patients with isolated ostial/midshaft lesions of ULMCA who received PCI or CABG in our hospital from 2003-01 to 2009-07 were enrolled.The patients were divided into 2 groups:PCI group,n=149 patients who received drug-eluting stents (DES) implantation and CABG group,n=110.The end points were all cause death,myocardial infarction (MI) and major adverse cardiac and cerebrovascular events (MACCE) which included cardiac death,non-fetal MI,stroke,repeated revascularization and the composite events death.Results:The median follow-up period was 7.1 years (inter quartile range 5.3-8.2 years) in all patients.Before multivariate adjusting,the following parameters were similar between PCI group and CABG group:all cause death (12.7％ vs 29.7％),P=-0.096;non-fatal MI (14.8％ vs 8.5％),P=0.844;stroke (9.3％ vs 6.3％),P=0.904;repeated revascularization (26.8％ vs 19.0％),P=0.234;composite events of cardiac death/stroke/MI (18.9％ vs 20.3％),P=0.224 and MACCE occurrence (37.5％ vs 34.2％),P=0.946.With adjusted variations,the trend was similar to pre-adjustment.Conclusion:During the maximum 8.2 years follow-up period,PCI and CABG had the similar efficacy and safety in patients with ostial/shaft lesions of ULMCA.

Background: Esophageal cancer is associated with substantial disease burden in China, and data on the economic burden are fundamental for setting priorities in cancer interventions. The medical expenditure for the diagnosis and treatment of esophageal cancer in China has not been fully quantified. This study aimed to examine the medical expenditure of Chinese patients with esophageal cancer and the associated trends. Methods: From 2012 to 2014, a hospital-based multicenter retrospective survey was conducted in 37 hospitals in 13 provinces/municipalities across China as a part of the Cancer Screening Program of Urban China. For each esophageal cancer patient diagnosed between 2002 and 2011, clinical information and expense data were extracted by using structured questionnaires. All expense data were reported in Chinese Yuan (CNY; 1 CNY= 0.155 USD) based on the 2011 value and inflated using the year-specific health care consumer price index for China. Results: A total of 14,967 esophageal cancer patients were included in the analysis. It was estimated that the overall average expenditure per patient was 38,666 CNY, and an average annual increase of 6.27％ was observed from 2002 (25,111 CNY) to 2011 (46,124 CNY). The average expenditures were 34,460 CNY for stage Ⅰ, 39,302 CNY for stage Ⅱ, 40,353 CNY for stage Ⅲ, and 37,432 CNY for stage IV diseases (P < 0.01). The expenditure also differed by the therapy type, which was 38,492 CNY for surgery, 27,933 CNY for radiotherapy, and 27,805 CNY for chemotherapy (P < 0.05). Drugs contributed to 45.02％ of the overall expenditure. Conclusions: These conservative estimates suggested that medical expenditures for esophageal cancer in China substantially increased in the last 10 years, treatment for early-stage esophageal cancer costs less than that for advanced cases, and spending on drugs continued to account for a considerable proportion of the overall expenditure.

OBJECTIVETo investigate the effect of premature rupture of the membrane (PROM) on neonatal complications in premature infants.

METHODSThe registration information of 7684 preterm infants with gestational age <37 weeks were collected from the cooperative units in the task group between January 1, 2014 to December 31, 2014. Specially trained personnel from each cooperative units filled in the unified form in a standardized format to record the gender, gestational age, birth weight, PROM, placental abruption, antenatal corticosteroid, Apgar score, amniotic fluid pollution, and complications of the infants. The data were analyzed comparatively between the cases with PROM and those without (control).

RESULTSThe preterm mortality rate was significantly lower but the incidences of ICH, NEC, ROP and BPD were significantly higher in PROM group than in the control group (P<0.05). The 95% confidence interval of the OR value was <1 for mortality, and was >1 for ICH, NEC, ROP and BPD. After adjustment for gestational age, birth weight, gender, mode of delivery, placental abruption, placenta previa, prenatal hormones, gestational diabetes mellitus (GDM), gestational period hypertension and 5-min Apgar score <7, the incidences of NEC, ROP and BPD were significantly different between the two groups (P<0.05) with 95% confidence interval of OR value >1, but the mortality rate and incidence of ICH were not significantly different between the two groups (P>0.05).

CONCLUSIONPROM is a risk factor for NEC, ROP and BPD in preterm infants, and adequate intervention of PROM can reduce the incidences of such complications as NEC, ROP and BPD in the infants.

Apgar Score ; Birth Weight ; Female ; Fetal Membranes, Premature Rupture ; pathology ; Gestational Age ; Humans ; Incidence ; Infant, Newborn ; Infant, Newborn, Diseases ; etiology ; Infant, Premature ; Pregnancy ; Risk Factors