Bisphenol A induces testicular oxidative stress in mice leading to ferroptosis / 亚洲男科学杂志(英文版)

Bisphenol A is a common environmental factor and endocrine disruptor that exerts a negative impact on male reproductive ability. By exploring bisphenol A-induced testicular cell death using the Institute of Cancer Research (ICR) mouse model, we found that a ferroptosis phenomenon may exist. Mice were divided into six groups and administered different doses of bisphenol A via intragastric gavage once daily for 45 consecutive days. Serum was then collected to determine the levels of superoxide dismutase and malondialdehyde. Epididymal sperm was also collected for semen analysis, and testicular tissue was collected for ferritin content determination, electron microscope observation of mitochondrial morphology, immunohistochemistry, real-time quantitative polymerase chain reaction, and western blot analysis. Exposure to bisphenol A was found to decrease sperm quality and cause oxidative damage, iron accumulation, and mitochondrial damage in the testes of mice. In addition, bisphenol A was confirmed to affect the expression of the ferroptosis-related genes, glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), cyclooxygenase 2 (COX2), and acyl-CoA synthetase 4 (ACSL4) in mouse testicular tissues. Accordingly, we speculate that bisphenol A induces oxidative stress, which leads to the ferroptosis of testicular cells. Overall, the inhibition of ferroptosis may be a potential strategy to reduce male reproductive toxicity caused by bisphenol A.

The role of SIRT6 in nonalcoholic steatohepatitis / 生理学报

SIRT6, a member of the silencing information regulatory protein family, is a nicotinamide adenine dinucleotide-dependent histone deacetylase and an ADP-ribose transferase enzyme. It plays an important role in fundamental physiological and pathological processes, including lipid metabolism, inflammation, oxidative stress and fibrosis, and is considered as a potential therapeutic target for metabolic syndrome. SIRT6 knockout mice displayed severe fatty liver, and the expression of SIRT6 in the liver of nonalcoholic steatohepatitis (NASH) mice was significantly lower than that of normal mice. Overexpression of SIRT6 significantly ameliorated NASH-induced liver damage. It is suggested that SIRT6 may play a key role in protecting against NASH. In this paper, we review the important regulatory functions of SIRT6 in the occurrence and development of NASH.

Effect of Aloesin on Proliferation and Apoptosis of Non-small Cell Lung Cancer A549 Cells / 中国实验方剂学杂志

Objective:To study the mechanism of aloesin in inducing apoptosis in human non-small cell lung cancer (NSCLC) A549 cells, so as to inhibit its proliferation. Method:A549 cells in logarithmic growth phase were collected, and cell counting kit-8 (CCK-8) was used to detect the effect of different concentrations of aloesin (2, 4, 8, 16, 32, 64, 128 μmol·L-1) on the proliferation of A549. Effect of aloesin (0, 16 μmol·L-1) on the number of clones formed in A549 cells and the size of clone formation was determined by crystal violet staining. effect of aloesin on apoptosis of A549 cells was detected by annexin V/propidium iodide(PI)apoptosis kit staining. Hoechst staining was used to detect the phenomenon of apoptotic nuclear pyknosis. Western blot was used to detect aloesin's effect on death-related protein expressions of Bcl-xl/Bcl-2 associated death promoter (Bad), cleaved-Caspase-3,cl-Caspase-3(Asp175), Caspase-3, cleaved poly ADP-ribose polymerase (cl-PARP), poly ADP-ribose polymerase (PARP) in A549 cells. In vivo, 5-week-old nude mice were subcutaneously inoculated with 2×106 A549 cells, and randomly divided into the medication group and the blank group. aloesin or normal saline was intraperitoneally injected for 4 weeks, and the tumor volume of nude mice was measured weekly. The body weight of the mice was observed, and the appearance of the nude mice was observed. Result:Aloesin inhibited the proliferation and cloning of A549 cells in a concentration-dependent manner (PPPPPPin vivo, aloesin significantly shrank the volume of subcutaneous tumors in mice, reduced tumor weight, with a better appearance than that of the control group. Conclusion:Aloesin may inhibit the expression of NSCLC by inducing apoptosis of A549 cells, and is safe to use, with no inhibitory effect on the body weight of mice.

Inhibitory Effect of Zeqi Tang on Mouse Model of Lung Cancer / 中国实验方剂学杂志

Objective:To observe effect of Zeqi Tang in intervening mice with orthotopic lung cancer model, in order to observe its anti-tumor mechanism. Method:An in situ mouse model of non-small cell lung cancer was established through intrapulmonary injection with 1×105 LLC-luc cells. The model mice were intragastrically administered with Zeqi Tang(0.171 g·mL-1) or normal saline for 35 days. Appearance (spirit, hair, appetite, sleep), survival period and Zeqi Tang anti-tumor effect were observed, weekly vital imaging was performed to detect the fluorescence signal in the lungs of mice. Flow cytometry was used to detect the NK cell content in the spleen of the model mice. CD107α was used to detect the degranulation of NK cells in the spleen of mice after administration of Zeqi Tang. Kromasil 100 5 C18 column was used and eluted with acetonitrile-0.025%phosphoric acid in a gradient mode, with flow rate at 1.0 mL·min-1, column temperature at 35℃ and detection wavelength of 265 nm, as to establish the fingerprint of Zeqi Tang. The fingerprints of 10 batches of samples was evaluated by using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition) recommended by the Chinese Pharmacopoeia Commission, in order to complete the quality control of Zeqi Tang. Result:Zeqi Tang could significantly inhibit the lung fluorescence signal of lung cancer in situ model mice and prolong the survival of mice(PPPα also increased significantly(PConclusion:Zeqi Tang may enhance the tumor growth and prolong the survival period of mice by up-regulating the number of NK cells in mice and enhancing their degranulation function. The evaluation of similarity of HPLC fingerprint of Zeqi Tang reflects the quality of lacquer soup to a certain extent, and can provide reference for further study.

Study of the therapeutic effect of oral propranolol combined with topical timolol maleate in the treatment of mixed maxillofacial hemangioma in infants / 口腔疾病防治

Objective@#To observe the treatment effect of oral propranolol combined with topical timolol maleate for infantile maxillofacial mixed hemangioma and provide evidence for clinical treatment.@*Methods@# Ninety-seven cases of infantile maxillofacial mixed hemangioma were enrolled. The cases were randomly divided into A and B groups: 50 cases in group A were treated with oral propranolol combined with topical timolol maleate, and 47 cases in group B were treated with oral propranolol only. The changes in the color, volume, and texture of the tumors were recorded before and after treatment, and color ultrasonography of the lesion area was performed. The follow-up time was 1-12 months. The differences in the curative effect, effective time and adverse reaction between the two groups were compared. @*Results @#The effective rate of group A was 92.0% (46/50) and that of group B was 74.5% (35/47), with a statistical significance (P < 0.05). The mean time of treatment in group A was 4.2 months and that in group B was 5.5 months. Compared with group B, the treatment time of group B was shorter (t=3.211, P < 0.05), and no serious adverse reactions occurred in both groups.@*Conclusion@#Oral propranolol combined with topical timolol maleate is effective in the treatment of mixed hemangioma of the maxillofacial region in infants.

Cyanidin-3-glucoside Inhibits Platelet-Derived Chemokines and Their Receptors on Leukocytes of ApoE-/-mice / 中山大学学报(医学科学版)

[Objective]To investigate the effects of different doses of anthocyanins Cy-3-g on serum lipids,platelet-derived chemokines,including MIF and CXCL12 together with their receptors CXCR4 and CXCR7.[Methods]Male ApoE-/-mice were distributed to 5 groups(n=15 per group),and fed with standard diet,high fat diet(HFD),HFD with 200 mg/kg Cy-3-g(low),HFD with 400 mg/kg Cy-3-g(middle),HFD with 800 mg/kg Cy-3-g(high)respectively for 16 weeks. The changes of body weight and food intake were recorded weekly. At the end of the experiment term,the serum lipids(triglyceride,cholesterol,HDL-C,LDL-C)were detected by kits. Arteries were separated to determine plaque histology by Oil-Red-O stain.MIF,CXCL12 and CCL2 in serum were detected by ELISA kits.The expression of CXCR4 and CXCR7 on the surface of leukocytes were tested via flow cytometry. Tail bleeding time was measured mean-while.[Results]Compared with the HFD group,the levels of serum lipids in medium(400 mg/kg)and high(800 mg/kg)Cy-3-g groups were significantly decreased(P<0.05). The plaque area of carotid artery was decreased in high Cy-3-g group(P<0.05).Cy-3-g at all doses significantly reduced the serum concentrations of CXCL12 and CCL2(P<0.002).Cy-3-g of medium(400 mg/kg)and high(800 mg/kg)dose significantly inhibited the expression of CXCR4 and CXCR7 on leukocyte surface(P<0.05). Cy-3-g does not prolong the tail bleeding time.[Conclusions]Anthocyanin Cy-3-g inhibits chemokine CXCL12,CCL2 in serum,and the expression of CXCR4 and CXCR7 on leukocytes without bleeding risk in ApoE-/-mice.

Cyanidin-3-o-β-glucoside Inhibits THP-1 Cells Migration Mediated by Platelet Derived Chemotactic Factor / 中山大学学报(医学科学版)

[Objective]To evaluate the effects of anthocyanin cyanidin-3-o-β-glucoside(Cy-3-g)on monocyte migra-tion mediated by platelet derived factors in vitro.[Methods]Thrombin-activated human gel-filtered platelets were incubated with different concentrations(0,0.5,5 or 50 μmol/L)of Cy-3-g.The level of TGF-β1,β-TG,CCL5 in platelet superna-tant was determined by ELISA.The peripheral venous blood from healthy volunteers were incubated with different concentra-tions of Cy-3-g. The expression of CCR5 on leukocytes was detected by flow cytometer. Calcein AM labeled THP-1 cells were incubated with the releasates of Cy-3-g-treated gel-filtered platelets for 120 min,and then the number of the THP-1 cells were counted by a microscope.Moreover,THP-1 cells were pre-incubated with different concentrations of Cy-3-g with or without CCR5 inhibitor maraviroc(200 μmol/L)for 60 min,then treated with recombinant CCL5(100 nmol/L)for 120 min.The number of the THP-1 cells were counted as well.[Results]Cy-3-g significantly inhibited platelet TGF-β1,β-TG,CCL5 secretion and decreased CCR5 expression of leukocytes compared with the control(P<0.05).Moreover,the sig-nificant inhibitory effects of Cy-3-g on THP-1 cell migration toward the releasates of Cy-3-g-treated platelets were also ob-served(P<0.05). In addition,THP-1 cell migration toward recombinant CCL5 was significantly suppressed by Cy-3-g.[Conclusions]Anthocyanin Cy-3-g inhibited inflammation of atherosclerosis by platelet TGF-β1,β-TG,CCL5 secretion and CCL5-mediated monocyte migration.Our results provided new ideas for prevention and treatment of atherosclerosis.

An investigation on the antibody levels to pertussis and diphtheria among healthy people in Wenzhou City / 预防医学

Objective To learn the immunity levels of pertussis and diphtheria among healthy people in Wenzhou City,and to provide scientific evidence for the effective control of the two diseases.Methods Stratified sampling method was applied in this investigation and 1 350 healthy people were investigated and provided serum samples.Vaccine immunization of pertussis and diphtheria and demographic characteristics were also collected.Enzyme linked immunosorbent assay (ELISA)was conducted to detect pertussis and diphtheria antibodies.Those with more than 1 00 IU/ml aged more than 3 years were also investigated the disease history.The estimated infection rate of pertussis for population aged more than 3 years was based on the test results.The attenuation trend of pertussis and diphtheria immunity levels after vaccination was analyzed contrastively.Results The antibody positive rate to pertussis was 36. 52%,and the median of antibody concentration was 1 9. 45 IU/m1 .The antibody positive rate and concentration was highest among 36 -60 years old people (64. 29%,36. 39 IU/ml ).The vaccination rate of population with more than 3 doses of DPT (Diphtheria Toxoid -Pertussis Vaccine-Tetanus Toxoid)was 95. 80%,and the corresponding positive rate was 24. 36%.The positive rate was 28. 57% in 0-3 month and 1 0. 71% in 1 0 -1 2 month after vaccination.The antibody concentration to pertussis had a negative correlation with days after vaccination(r=-0. 22,P<0. 05).The proportion of subjects with more than 1 00 IU/ml in population aged more than 3 years was 7. 91%.The estimated infection rate of pertussis for population aged more than 3 years was 49. 27%.The antibody positive rate to diphtheria was 96. 00%,and the median of antibody concentration was 0. 1 3 IU/m1 . The positive rate was highest (1 00%) among 1 -2 years old people and lowest (82. 5%) among newborns.Antibody positive rate (protective rate)and antibody concentration to diphtheria of population which had more doses of DPT or had vaccinated with DT were higher than those had not (P<0. 05 ).The sustainability of vaccine to diphtheria was higher than pertussis after vaccination of DPT.Conclusion Residents in Wenzhou are generally vulnerable to pertussis.Natural infection is considered to be an important influencing factor for the level of pertussis.The level of diphtheria antibodies in Wenzhou is high.The vaccine containing diphtheria is considered useful for children.However,it is suggested to conduct diphtheria booster immunization in older age group.

Surveillance analysis on anaphylactic rash after immunization in Ouhai District of Wenzhou City, 2008-2013 / 上海预防医学

[ Objective ] To analyze the characteristics of anaphylactic rash after vaccination in Ouhai District of Wenzhou City during 2008-2013. [ Methods ] Data on anaphylactic rash cases reported during 2008-2013 were collected through the national AEFI information management system.And descriptive epidemiologic methodology was used in this study. [ Results] A total of 111 anaphylactic rash cases were reported in Ouhai District during 2008-2013,the reported incidence rate of anaphylactic rash were 3.58 per million doses.The ratio of male-female was 1.27:1.Cases of ≤1 year old accounted for 65.77%.Those without fever accounted for 75.68%.The number of the reports for the third quarter of the year accounted for 41.44%of the total.The cases of anaphylactic rash mostly occurred within 24 h after immunization(81.08%) .The top three vaccines reported in occurrence of rash were measles and rubella at-tenuated live vaccine(35.14%), diphtheria, tetanus and acellular pertussis combined vaccine(18.92%), and A(H1N1)influenza vaccine(5.41%).The reported rates for the top three vaccines were 115.85, 29.33,13.07 per million doses for 7-valent pneumococcal polysaccharide conjugate vaccine,measles and rubella combined attenuated live vaccine, A(H1N1) influenza vaccine,respectively. [Conclusion] Differential diagnosis of anaphylactic rashes needs to be more stressed.Most anaphylactic rash were reported in the expected range,but still monitoring analysis on incidence of allergic rash should be enhanced.Vac-cines with higher incidence of rash reported should be further studied and analyzed.

An evaluation of immunization coverage of the first dose of measles containing vaccine using incidence rate / 预防医学

Objective To evaluate the immunization coverage of the first dose of measles containing vaccine (MCV1 )by using the incidence of measles in Wenzhou City.Methods Descriptive epidemiological methods were used to analyze measles cases that reported in Wenzhou city from 2007 to 2012 and evaluate the immunization coverage of the first dose of measles containing vaccine.Results The average annual incidence rate was 10.46/100 000 from 2007 to 2012,and the annual incidence rate was 43.44/100 000 for children aged from 8 months to 83 months (42.59%).Based on the proportion of immunized measles cases vaccine effectiveness (VE)of MCV,the evaluated coverage rate of MCV1 was 73.80% (VE=90%)or 84.92% (VE=95%)in children aged from 13 to 83 months.The evaluated coverage rate of MCV1 was 83.25%(VE=90%)or 90.86%(VE=95%)in local children and 69.5 1%(VE=90%)or 82.02%(VE=95%)in migrating children.The timely immunization rate of MCV1 was 59.48% (VE =90%)or 74.59% (VE =95%).Conclusion The coverage rate and timely coverage rate of MCV1 are still low.It is important to strengthen the management of migrating population and enhance propaganda to ensure a high level vaccination rate to accelerate the elimination of measles.

Investigation on the natural infectious status of hantaviruses among small mammals in Longquan city, Zhejiang province / 中华流行病学杂志

Objective To investigate the situation of the natural infection of hantaviruses (HV) in small mammals and to provide evidence for the control and prevention of hemorrhagic fever with renal syndrome (HFRS) in Longquan area,Zhejiang province. Methods Small mammals were captured by night trap, and lung tissue samples were collected and stored in liquid nitrogen. HV antigens were detected by indirect immuno-fluorescence assay (IFA). The partial S genome segment sequences were amplified by RT-PCR. DNAStar program was used for editing and comparing the sequences. Phylogeny was analyzed through PAUP*4.0 software. Results 319 small animals were collected in Longquan, and 9 hantavirus antigen-positive samples were identified. The positive rate of hantavirus in Apodemus agrarius was 4.97% . Phylogenetic tree constructed by partial S segment (620-999 nt) showed that the 9 strains carried by A agrarius from Longquan all belonged to HTNV,and had a closer evolutionary relationship with isolate Z251 from Zhejiang province. Conclusion Our results indicated that the main host was A. agrarius and the infection rate of HTNV was high in Longquan area.

Genetic analysis of hantaviruses carried by Rattus norvegicus collected from Henan and Neimenggu provinces, China / 中华流行病学杂志

Objective Genetic analysis was performed to infer the relationship between hantaviruses carried by Rattus norvegicus from Henan and Neimenggu provinces and the other known hantavirus and the vaccine strain. Methods Total RNA was extracted from lung tissues with Trizol reagent. The complete M and S segment sequences of strains NM133 and Q12 were amplified by RT-PCR. The purified DNA fragments were directly subjected to sequencing, and then to sequence analysis and phylogenetic analysis. Results The complete S segment sequences of strains NM133 and Q12 were found to be 1770 nt and 1772 nt in length respectively, with one open reading frame encoding 429 amino acids. The complete M segment sequences of both two strains are 3654 nucleotide in length encoding a protein of 1133 amino acids. The two strains shared a high degree of homology with most of known Seoul virus (SEOV) but quite different from Hantaan virus and other hantaviruses. Furthermore, the nucleoprotein and glycoprotein of the two strains had the congruent structure with the vaccine strain Z37. On the S- and M-phylogenetic trees, both strains (NM133 and Q12) were grouped into the first cluster of SEOV, and were more closely related to the strains, such as: Hb8610, R22, HB55, L99, and K24-e7. Conclusion Both strains (NM133 and Q12) belonged to SEOV, and sharing a high degree of homology and similar secondary structure with strains including the vaccine strains Z37, our data suggested that the present vaccine used in China could effectively prevent HFRS caused by SEOV.

Genetic characterization of two Seoul virus strains isolated from patients and rats in Hebei province / 中华流行病学杂志

Objective Complete S and M segments of two Seoul virus (SEOV) strains were obtained to determine their genetic types and characteristics. Methods The complete S and M segments from the isolate Li and lung tissue (sample LF18) were amplified by RT-PCR. Genetic analyses were performed by using DNAStar and PHYLIP program package. Results Their sequences consisted of 1772 nucleotides, and had an open reading frame (ORF, 43 to 1332 nt)encoding a nucleoprotein of 429 amino acids for both two strains. The complete M segment sequences consisted of 3653 nucleotides and had an ORF encoding a GnGc precursor of 1133 amino acids. The GnGc precursor of the two strains had 62 cysteine and 6 N-glycosylation sites. Both two strains shared a high degree of homology with other known SEOV strains including strains L99, Gou3, and vaccine strain Z37, with 87.6% to 99.2% and 83.6% to 97.3% nucleotide identities, respectively. On the S-and M- trees, the two strains LF18 and Li were grouped into the third cluster of SEOV. Conclusion Both LF18 and Li strains belonged to SEOV and shared the congruent genetic characteristics with the vaccine strains Z37. Thus, the bivalent vaccine including the strain Z37 could effectively prevent HFRS which was caused by SEOV in Hebei province.

The molecular epidemiologic investigation of hantavirus carried by rodent hosts in Wenzhou, Zhejiang province / 中华流行病学杂志

Objective To study the epidemiological features of hantavirus in rodents in Wenzhou,Zhejiang province.Methods Rodents were captured in Wenzhou,where hemorrhagic fever with renal syndrome (HFRS) had been endemic. Hantavirus antigens in the rat lungs were detected by immunofluorescence assay (IFA).Partial S segment (nt 620-999) and partial M segment (nt 2001-2301)sequences were amplified by RT-PCR,and then sequenced.Neighbor-joining method was used to construct for phylogenetic analysis.Results A total of 96 rodents were trapped in the epidemic areas,and 6 hantavirus antigens were identified from these lung samples (6.3%).Partial S and partial M segment sequences were successfully recovered from 5 samples and determined. Phylogenetic analysis of these sequences indicated that all viruses belonged to Seoul virus (SEOV),regardless of the sources (Rattus norvegicus,Rattus tanezumi and Rattus rattoide) that they were derived. However,the clustering pattern in the partial S-tree was different from that in the partial M-tree,suggesting that the re-assortment between SEOVs had occurred.Conclusion All Rattus rats carried SEOV in Wenzhou and the genetic reassortment with SEOV had occurred naturally.