Ketosis is an energy metabolism disorder occurring frequently in periparturient dairy cows,primarily attributed to elevated non-esterified fatty acid(NEFA)levels resulting from nega-tive energy balance(NEB).Excessive NEFA will be incompletely oxidated into large amounts of ketone bodies or be re-esterified and deposit in the liver as a consequence of hepatic limited oxida-tive capacity,ultimately leading to ketosis and fatty liver.Hypoxic microenvironments are com-monly found during the progression of various liver diseases.Hypoxia inducible factor-2 alpha(HIF-2 alpha)has been identified as a crucial regulator of lipid metabolism.However,it is still un-clear the association between HIF-2α and disrupted lipid metabolism in the livers of in ketotic cows.This study aims to investigate the effect of high concentrations of NEFA on HIF-2α expres-sion and cellular oxygen homeostasis through bovine liver tissue and primary hepatocytes.In vivo,hepatic triglyceride(TAG)content was assessed to determine the extent of hepatic lipid accumula-tion,and HIF-2α protein and mRNA levels were analyzed by immunohistochemistry staining,Western blot and qRT-PCR assay in liver tissue samples from dairy cows;in vitro,bovine primary hepatocytes were treated with different concentrations of NEFA.Oil Red O staining and TAG con-tent assay were performed to determine hepatocellular steatosis extent,and immunofluorescence staining.Western blot,and qRT-PCR were performed to analyze HIF-2α expression,in addition,lu-minescent oxygen sensor[Ru(dpp)3]Cl2 was added to indicate intracellular oxygen levels.These results showed a significant increase in TAG content and elevated HIF-2α expression in the liver tissue of ketotic cows,and high concentrations of NEFA induced lipid accumulation,upregulation of HIF-2α expression,and intracellular hypoxia in bovine primary hepatocytes.These findings sug-gested that HIF-2α was significantly"activated"in the liver of ketotic cows and high concentration of NEFA-induced bovine primary hepatocytes,and that high concentrations of NEFA induced in-tracellular hypoxia in vitro.This study provides a potential molecular target for further investiga-tion of the mechanism underlying hepatic lipid metabolism disorders in ketotic cows.

Ketosis is an energy metabolism disorder occurring frequently in periparturient dairy cows,primarily attributed to elevated non-esterified fatty acid(NEFA)levels resulting from nega-tive energy balance(NEB).Excessive NEFA will be incompletely oxidated into large amounts of ketone bodies or be re-esterified and deposit in the liver as a consequence of hepatic limited oxida-tive capacity,ultimately leading to ketosis and fatty liver.Hypoxic microenvironments are com-monly found during the progression of various liver diseases.Hypoxia inducible factor-2 alpha(HIF-2 alpha)has been identified as a crucial regulator of lipid metabolism.However,it is still un-clear the association between HIF-2α and disrupted lipid metabolism in the livers of in ketotic cows.This study aims to investigate the effect of high concentrations of NEFA on HIF-2α expres-sion and cellular oxygen homeostasis through bovine liver tissue and primary hepatocytes.In vivo,hepatic triglyceride(TAG)content was assessed to determine the extent of hepatic lipid accumula-tion,and HIF-2α protein and mRNA levels were analyzed by immunohistochemistry staining,Western blot and qRT-PCR assay in liver tissue samples from dairy cows;in vitro,bovine primary hepatocytes were treated with different concentrations of NEFA.Oil Red O staining and TAG con-tent assay were performed to determine hepatocellular steatosis extent,and immunofluorescence staining.Western blot,and qRT-PCR were performed to analyze HIF-2α expression,in addition,lu-minescent oxygen sensor[Ru(dpp)3]Cl2 was added to indicate intracellular oxygen levels.These results showed a significant increase in TAG content and elevated HIF-2α expression in the liver tissue of ketotic cows,and high concentrations of NEFA induced lipid accumulation,upregulation of HIF-2α expression,and intracellular hypoxia in bovine primary hepatocytes.These findings sug-gested that HIF-2α was significantly"activated"in the liver of ketotic cows and high concentration of NEFA-induced bovine primary hepatocytes,and that high concentrations of NEFA induced in-tracellular hypoxia in vitro.This study provides a potential molecular target for further investiga-tion of the mechanism underlying hepatic lipid metabolism disorders in ketotic cows.

Abstract: Objective　To identify the species of Mycobacteroides abscessus complex (MABC) in patients with pulmonary infection from the Second Affiliated Hospital of Hainan Medical University, and to investigate the species types, drug sensitivity and population distribution of MABC in pulmonary infection in Hainan. Methods　Respiratory tract specimens were collected from suspected tuberculosis patients who visited the Second Affiliated Hospital of Hainan Medical University from January 2014 to December 2021 and cultured for Mycobacterium isolation. Non-tuberculous mycobacteria (NTM) strains were preliminarily identified by p-nitrobenzoic acid/thiophen-2-carbohydrazide (PNB/TCH) medium and DNA microarray chip, and then MABC and its subspecies were identified by hsp65 and rpoB gene sequencing. In vitro antimicrobial susceptibility test was performed by broth microdilution method. Results　A total of 3 025 respiratory specimens from suspected pulmonary tuberculosis patients were collected during the study period. Among the 123 patients with identified MABC isolates, 124 MABC strains were isolated and identified, including 74 strains of Mycobacteroides abscessus subsp. abscessus, 38 strains of Mycobacteroides abscessus subsp. massiliense and 12 strains of Mycobacteroides abscessus subsp. bolletii. Among them, 118 patients had single MABC subspecies infection, one patient had mixed infection with two MABC subspecies, two patients had mixed infection with MABC and other NTM, and two cases had mixed infection with MABC and M.tuberculosis. There were more female patients than male patients with a ratio of 1:0.64, and those aged 50 and above amounted to 76.42% (94/123, 95%CI: 67.93%-83.61%). There was no significant difference in age distribution between male and female patients (Z=-0.944, P=0.347). The drug susceptibility results showed that all MABC strains were sensitive to Tigecycline (TGC), with a resistance rate of 0.81% (1/124) to Amikacin (AK), and resistance rates of 6.45% (8/124), 32.26% (40/124), and 74.19% (92/124) to Cefoxitin (FOX), Linezolid (LZD), and Imipenem (IPM), respectively. For Clarithromycin (CLR), MABC showed induced resistance , and there was a statistically significant difference in the CLR (14D) resistance rates among the three subspecies (χ2=66.335, P<0.001). The resistance rates to Tobramycin (TOB), Doxycycline (DOX), Moxifloxacin (MFX), Ciprofoxacin (CIP), Trimethoprim/Sulfamethoxazole (TMP-SMX), and Amoxicillin/Clavulanic acid (AMC) were high, all >80%. Conclusion　 In Hainan Province, pulmonary infections with MABC are mainly caused by Mycobacteroides abscessus subsp. Abscessus, which show high rates of inducible resistance to CLR. Timely and accurate identification of MABC to subspecies and drug susceptibility testing are of significant important for clinical decision-making.

Objective:To analyze spatiotemporal distribution characteristics of syphilis in Zhejiang province from 2016 to 2020, to explore cluster areas of syphilis cases, and to provide a theoretical basis for accurate prevention and control of syphilis.Methods:Data on reported syphilis cases in Zhejiang province from 2016 to 2020 were collected from the notifiable infectious disease surveillance system in China Disease Control and Prevention Information System. The ArcGIS10.2 software was used as a data management and presentation platform to establish a database for spatial analysis of syphilis in Zhejiang province from 2016 to 2020, and spatial autocorrelation analysis of reported syphilis incidence was conducted. The SaTScan 9.6 software was used for spatiotemporal scanning analysis.Results:A total of 158 420 cases of syphilis were reported in Zhejiang province from 2016 to 2020, and the average annual reported incidence rate was 49.07 per 100 000 in all counties and districts (range: 20.52 per 100 000-124.29 per 100 000) . The overall spatial distribution pattern of syphilis cases in Zhejiang province was characterized by higher reported syphilis incidence in the West area and lower incidence in the middle area. Global autocorrelation analysis showed that all the global Moran′s I indices from 2016 to 2020 were over 0, Z values were over 1.96, and P values were below 0.001, suggesting the spatial clustering of reported syphilis cases. Local autocorrelation analysis showed that there were 2 high-high clustering areas (Tonglu and Chun′an counties in Hangzhou city, Jingning county in Lishui city) , and 1 low-low clustering area (Jindong district, Dongyang and Yongkang county-level cities in Jinhua city) . Local hot-spot analysis with G statistic showed that there were 6 positive hot-spot areas and 8 negative hot-spot areas of syphilis cases in Zhejiang province. SaTScan spatiotemporal scanning analysis revealed 3 clustering areas, mainly distributed in Taizhou and Zhoushan cities along the eastern coast, and some counties and districts in the southwestern mountainous areas. Conclusions:The reported incidence rate of syphilis in Zhejiang province is characterized by spatial clustering. In the future, prevention and control of syphilis should be strengthened in southwestern mountainous areas, eastern islands, and other remote areas as well as areas with poor transportation.

Objective: To assess the implementation of National Leprosy Control Plan (2011-2020) in Zhejiang Province, so as to provide insights into leprosy control. Methods: Leprosy control data were collected from the National Leprosy Management Information System of China, and health administrative sectors, centers for disease control and prevention and professional leprosy control institutions at each level in Zhejiang Province. According to the scheme for the final evaluation of National Leprosy Control Plan (2011-2020), the implementation of 12 indicators in Zhejiang Province in 2020 was investigated, including the current number of leprosy patients, prevalence of leprosy, proportion of training on leprosy control skills, proportion of regular leprosy treatment, proportion of treatment of adverse reactions, annual examination rate of close contacts, proportion of early case identification and awareness of leprosy control knowledge. Results: There were 50 registered leprosy cases in Zhejiang Province by the end of 2020, with a decrease of 50.98% relative to in 2010. The prevalence of leprosy was less than 1/105 in 93 counties (districts) of Zhejiang Province, and the rate of training on leprosy control skills, rate of regular leprosy treatment, rate of treatment of adverse reactions, annual examination rate of close contacts, rate of early case identification were all 100% in Zhejiang Province. There were no new confirmed cases with diagnosis of leprosy having grade 2 deformity or disability, or no new cases with deformity or disability within 2 years following anti-leprosy therapy. In addition, the awareness of leprosy control knowledge was 91.67% among the public and 98.12% among the close contacts. All of the 12 indicators reached the requirements of the National Leprosy Control Plan (2011-2020). Conclusions The implementation of the National Leprosy Control Plan (2011-2020) in Zhejiang Province had achieved the targets defined in the final evaluation of the plan. Intensifying multi-sectoral joint leprosy prevention and control and improving early identification and standardized therapy of leprosy cases are recommended for future leprosy control in Zhejiang Province.

Objective:To investigate the epidemiological characteristics of syphilis in Zhejiang province and to provide scientific basis for the development of syphilis prevention and control strategies.Methods:A descriptive epidemiological analysis was conducted on the incidence data of syphilis in Zhejiang from 2010 to 2019.Results:During the period, the incidence rate of syphilis decreased from 94.90/100 000 in 2010 to 53.53/100 000 in 2019 with an average decreasing rate of 6.16%. The annual decreases of the incidences of congenital syphilis, primary syphilis and secondary syphilis were all obvious, which were 43.47%, 21.38% and 14.19% respectively. The proportion of latent syphilis cases increased with year. Except for Lishui, the incidences of syphilis in the remaining 10 prefectures showed declining trends. The incidence rates in both men and women showed declining trends with the average rates of 4.80% and 6.45% respectively. The incidence peaks occurred in old men aged ≥60 years and in sexually active women aged 20-34 years, and the syphilis cases in age group ≥60 years increased significantly. The cases were mainly farmers, accounting for 43.00%.Conclusion:The incidence of syphilis in Zhejiang showed a decreasing trend, but the situation remains serious, indicating that the intensity and quality of the comprehensive prevention and control needs to be further strengthened.

Objective: To investigate the effects of exposing Trichophyton rubrum fungus to microwaves at different intensities in terms of the activity of succinic dehydrogenase and beta-(1, 3)-D-glucan synthase. Methods: Trichophyton rubrum organisms were randomly divided into a control group and experimental groups. The experimental groups were incubated at 27 ℃ after direct radiation with 2450 MHz microwaves at 20, 40, 60 or 80 W for 15 min, repeated 8 times. The control group was incubated without any irradiation. Thirty days later, the beta-(1, 3)-D-glucan synthase and succinate dehydrogenase activities were determined using enzyme-linked immunosorbent assays. Results: The enzymatic activity decreased gradually with increasing radiation intensity. When the output power was 80 W, the beta-(1, 3)-glucan-synthase-D activity was 0.730±0.74 U/ml and that of the succinate dehydrogenase was 1.828±1.774 U/L, both significantly lower than in the groups subjected to less powerful irradiation. Conclusions Microwave radiation can decrease the enzymatic activity of Trichophyton rubrum in a dose-dependent manner. Higher intensity is more effective. Microwave irradiation can decrease the activity of succinate dehydrogenase and beta-(1, 3)-glucan synthase from Trichophyton rubrum in vitro, resulting in the destruction of fungal cell walls and interfering with the tricarboxylic acid cycle, furthering cell death. Moreover, the temperature change possibly also helps promote the biological effects of microwave radiation.

Objective To investigate the value of leukotriene B4(LTB4)and leukotriene E4(LTFA)in early diagnosis of renal involvement in children with Henoch Schonlein purpura(HSPN)and its influence on prognosis.Methods A total of 185 children with HSPN were enrolled in our hospital from January 2014 to October 2016.A total of 50 healthy children were selected as control group at the same period.The serum levels of LTB4 and LTFA in all subjects were detected,and their value in early diagnosis of HSPN and its influence on prognosis were analyzed.Results Compared with the control group,the serum levels of LTB4 and LT-FA in children with HSPN were significantly higher(P<0.05),the contents were 987.6 ng/L and 896.3 ng/L,respectively.The AUC of the combined detection of HSPN was 0.899 and the sensitivity was 0.810,which was better than that of LTB4 and LTB4 alone(P<0.05).The average follow-up was(28.4 ± 10.3)months,with grade A prognosis in 133 cases(71.9%),B grade in 47 ca-ses(25.4%),and C grade in 5 cases(2.7%).The rank sum test showed,the prognosis of children with LTB4<987.6 ng/L was better than that of children with LTB4≥987.6 ng/L,the prognosis of children with LTFA<896.3 ng/L was better than that of children with LTFA≥896.3 ng/L.Spearman analysis showed that the content of LTB4 and LTFA was negatively correlated with prognosis(Rs= -0.693 and -0.637,P<0.05).The higher contents of LTB4 and LTFA,the worse the prognosis.Conclusion Combined detection of LTB4 and LTFA has important clinical significance for early diagnosis and prognosis of HSPN.

Objective To develop a digital polymerase chain reaction (PCR) ribotyping method and database for Clostridium difficile genotyping.Methods Sequencer based fluorescence capillary gel electrophoresis was used,instead of agarose gel electrophoresis,to establish the digital PCR-ribotyping of Clostridium difficile.Forty Clostridium difficile reference strains,consisting of 10 PCR-ribotypes (RT),were genotyped by the new digital PCR-ribotyping method to set-up the database.Results The sequencer based fluorescence capillary gel electrophoresis correctly detected PCR-ribotyping products of the 40 reference strains,and showed as digital figure;significant differences of these digital figures were found between the 10 RT.High similar digital figures were shown in twenty-one RT027 strains,three RT002 strains and two RT014 strains.However,seven RT001 strains were typed as four subtypes,and two RT014 strains as two subtypes,respectively.Conclusion A digital PCR-ribotyping,and a reference database consisting of 10 RT are successfully established.

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .