Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.

Objective To analyze the distribution characteristics of traditional Chinese medicine(TCM)syndrome types in patients with osteoporosis and the distribution differences of clinical and serological indicators in TCM syndrome types.Meth-ods A total of 69 patients with osteoporosis were collected from the Third Affiliated Hospital of Guangzhou University of Chinese Medicine and Qifu Hospital Affiliated to Jinan University.The general information,bone mineral density T value,fasting periph-eral venous blood in the morning were collected.The expression of telomerase protective factor 1(POT1),telomerase reverse transcriptase(TERT),serum 8-hydroxy-2'-deoxyguanosine(8-OHdG)and superoxide dismutase 2(SOD2)were detected by ELISA.Finally,the above data were statistically analyzed.Results There were significant differences in body weight,height,bone mineral density,POT1,TERT,and 8-OHdG among the four syndromes(P＜0.05).In terms of correlation,the relation-ship between bone mineral density and each parameter in different syndrome types was explored.The bone mineral density of qi stagnation and blood stasis syndrome was positively correlated with SOD2 value.There is a positive correlation between bone min-eral density and 8-OHdG in patients with Yin deficiency of liver and kidney.TERT was positively correlated with qi stagnation and blood stasis syndrome.Liver and kidney Yin deficiency syndrome was positively correlated with weight and bone mineral den-sity,and negatively correlated with TERT value.Weight was negatively correlated with qi and blood stasis syndrome.Conclusion In TCM syndrome differentiation of osteoporosis,there were statistical differences in weight,height,bone mineral density,ser-um POT1,TERT and 8-OHdG among Qi-stagnation and blood stasis,spleen-kidney Yang deficiency,liver-kidney Yin deficiency and Qi-blood-peace syndrome.In different syndrome types,serum SOD2 and 8-OHdG were the influencing factors of bone miner-al density.Serum TERT and 8-OHdG are the main factors affecting the dialectical classification of osteoporosis.

The surveillance data of new cases of acute myocardial infarction (AMI) from January 1, 2013, to December 31, 2021, in Tengzhou City, Shandong Province, were used to analyze the incidence rate of AMI and its change trend among residents. The age and gender standardized incidence rate was calculated based on the 7th National Population Census 2020. The Cochran-Armitage trend test was used to analyze the trend of onset time and age. From 2013 to 2021, the crude and standardized incidence rate of total AMI in Tengzhou City declined from 130.07/100 000 and 161.12/100 000 to 76.15/100 000 and 72.77/100 000 ( Z=-13.785 and -20.822, both P<0.001). The crude and standardized incidence rates of males were higher than those of females. In 2016, males aged 45-54 years old and females aged 35-64 years old increased by 33.33%, 103.65%, 106.30%, and 95.75% compared to 2015, and the differences were statistically significant ( χ2=6.512, 4.965, 25.115, and 46.004, all P<0.05). The incidence rate of AMI in men aged<35 and 35-44 years old had an upward trend. From 2013 to 2021, the incidence rate of AMI decreased by 55.15% in urban areas and 36.59% in rural areas ( Z=-8.529 and -11.235, both P<0.001).

The surveillance data of new cases of acute myocardial infarction (AMI) from January 1, 2013, to December 31, 2021, in Tengzhou City, Shandong Province, were used to analyze the incidence rate of AMI and its change trend among residents. The age and gender standardized incidence rate was calculated based on the 7th National Population Census 2020. The Cochran-Armitage trend test was used to analyze the trend of onset time and age. From 2013 to 2021, the crude and standardized incidence rate of total AMI in Tengzhou City declined from 130.07/100 000 and 161.12/100 000 to 76.15/100 000 and 72.77/100 000 ( Z=-13.785 and -20.822, both P<0.001). The crude and standardized incidence rates of males were higher than those of females. In 2016, males aged 45-54 years old and females aged 35-64 years old increased by 33.33%, 103.65%, 106.30%, and 95.75% compared to 2015, and the differences were statistically significant ( χ2=6.512, 4.965, 25.115, and 46.004, all P<0.05). The incidence rate of AMI in men aged<35 and 35-44 years old had an upward trend. From 2013 to 2021, the incidence rate of AMI decreased by 55.15% in urban areas and 36.59% in rural areas ( Z=-8.529 and -11.235, both P<0.001).

Objective Analyze the temporal trend of ischemic stroke(IS)mortality among the residents of Tengzhou City of Shandong Province during the period of hypertension control from 2013 to 2021.Methods On January 1,2013,Tengzhou City,Shandong Province,began its hypertension control program.The IS mortality rate was calculated using the mortality data from January 1,2013 to December 31,2021,and analyzed for its time trend among residents with different characteristics.The registered population was derived from the Public Security Bureau of Tengzhou City,Shandong Province,and the age and sex standardized mortality rate was calculated using the data of China's 7th population census in 2020.The Chi-square test was used to compare the differences in mortality rate,and Cochran-Armitage trend test was used to compare the time trend and age trend of mortality rate.Results The overall crude and standardized mortality rates of IS in Tengzhou showed a temporal trend from 2013 to 2021(Z values were 12.647,7.305,respectively;all P＜0.001),and decreased by 23.77％and 30.99％(Z values were-7.393,-9.975,respectively;all P＜0.001)respectively in 2021 compared with 2019.The crude mortality rate of IS in male increased by 13.27％in 2019 compared with 2017,while the crude and standardized mortality rate in female decreased by 16.39％and 19.49％in 2018 compared with 2017,respectively,with statistical significance(x2 values were 7.160,9.789,and 15.109,respectively;all P＜0.05).Except the crude mortality rates in 2013 and 2015,the crude mortality rates and standardized mortality rates for males in other years were all higher than those for females,with statistically significant differences(x2 values:25.816-124.040,all P＜0.001).The crude mortality rate for IS increased with age in all years(Z values:42.604-61.025,all P＜0.001).The proportion of IS deaths among those aged≥65 was 85.85％.The overall crude mortality rates of the age group of male 45-54 years old showed a temporal trend from 2013 to 2021(Z=3.035,P＜0.01),while females in the same age group did not show a temporal trend(P＞0.05).The IS mortality rate in urban areas decreased from 62.61 per 100 000 in 2013 to 54.00 per 100 000 in 2021(Z=-2.097,P＜0.05).The rural areas increased by 213.15％in 2019 compared with 2013 and decreased by 22.75％in 2021 compared with 2019(Z values were 19.074,-6.390,respectively;all P＜0.001).Conclusions The IS mortality rate in Tengzhou City showed a decreasing trend in urban areas from 2013 to 2021,and a decreasing trend in rural areas after 2019.Compared to females,there is a trend of younger mortality among males in the age range of 45-54.Males and rural IS patients should be given special attention.

Objective To investigate the effects of galectin-3(Gal3)on skin abscess development and activation of mast cells(MC)in mice infected with methicillin-resistant Staphylococcus aureus(MRSA).Methods Wild type mice and Gal3-knockout(Gal3-/-)mice,at 6～8 weeks of age,were divided into four groups:Wild type mice+PBS group,Wild type mice+MRSA group,Gal3-/-mice+PBS group,Gal3-/-mice+MRSA group,were subcutaneously injected with MRSA or the same volume of phosphate buffer saline,with five mice per group.The development and pathological changes of skin abscess were monitored and recorded.The bacterial load in skin tissues was compared,and the expression of associated cytokines,degranulation of MC,and the distribution of MC activation marker 5-hydroxytryptamine(5-HT)were detected.Results The skin of Wild type mice showed progressive abscesses after subcutaneous infection with MRSA,but the Gal3-/-mice showed smaller abscess areas.Compared to the Wild type mice+MRSA group,the Gal3-/-mice+MRSA group showed lower bacterial loading in the skin tissues(P＜0.01)and fewer infiltrating inflammatory cells with histopathological observation.The expression of cytokines,including IL-1β,TNF-α,IL-33,TGF-β,and IL-10,were significantly lower in Gal3-/-mice than Wild type mice(P＜0.05).The toluidine blue staining showed a large number of degranulated MCs in the skin tissues of the wild type mice+MRSA group,whereas only a few degranulated MCs were observed in the Gal3-/-mice+MRSA group.It was further found that the expression of 5-HT in Gal3-/-mice+MRSA group was significantly lower than that in wild-type mice+MRSA group with immunohistochemical staining.Conclusion Gal3 deficiency reduced the activation and degranulation of mouse skin MC after MRSA infection,resulting in changes to inflammatory responses and alleviating the severity of skin tissue abscesses.

Objective:To analyze the impact of hypertension prevention and control on the mortality rate of acute myocardial infarction (AMI) in Tengzhou City, Shandong Province from 2013 to 2021.Methods:This study was a cross-sectional study. The surveillance data of AMI deaths from January 1, 2013 (the time when hypertension prevention and control began in Tengzhou) to December 31, 2021 were collected in the coronary heart disease information management system, the mortality rate of AMI and its change trend were analyzed, and the distribution differences among residents with different characteristics were analyzed. The registered population information was obtained from Tengzhou Public Security Bureau, and the age and gender standardized mortality rate was calculated based on the data of the 7th national population census in 2020. The t test was used to compare the differences in blood pressure and laboratory items, chi-square test was used to compare the differences in mortality rate, and Cochran-Armitage trend test was used to compare the time trend and age trend of mortality rate, so as to analyze the impact of hypertension prevention and control on the mortality rate of AMI. Results:The overall crude and standardized AMI mortality rates in Tengzhou decreased from 50.87/100 000 and 63.82/100 000 to 41.08/100 000 and 38.70/100 000 from 2013 to 2021, respectively ( Z=-5.741, -10.884, both P<0.001), and double peaks were formed in 2014 and 2017. The first peak of crude and standardized mortality rate was formed in 2015 for males, which was 25.12% and 17.60% higher than that in 2013; and the first peak was formed in 2014 for females, which was 29.56% and 24.38% higher than that in 2013 ( χ2=13.200, 9.065, 14.862, 12.123) (all P<0.05). The second peaks of crude and standardized mortality were formed in 2017, with an increase of 18.17% and 17.17% for males and 25.73% and 22.34% for females from 2016 ( χ2=8.266, 9.182, 14.066, 11.105), the standardized mortality rate was 15.18%-29.01% higher in males than that in females ( χ2=6.239-19.326) (all P<0.05). The mortality rate of AMI increased with age ( Z=35.485-51.308) ( P<0.001). Compared with 2013, the mortality rate in males aged 55 to 64 years in 2015 increased by 64.29% from that in 2013, and that of females in 2017 increased by 108.48% from that in 2015; and that in females aged 35 to 44 years in 2016 increased by 373.51% from that in 2015 ( χ2=10.751, 12.805, 4.799); in 2021, the age group of male and female≥65 years decreased by 43.51% and 41.28% when compared with that in 2013, respectively ( Z=-7.333, -7.465) (all P<0.05). The mortality rate of AMI in urban areas decreased by 76.93% in 2021 when compared with that in 2016, and in rural areas it decreased by 30.28% than that in 2017. Both regions showed a downward trend ( Z=-7.560, -2.398) (both P<0.05). Conclusions:The mortality rate of AMI in Tengzhou City, Shandong Province from 2013 to 2021 shows a decreasing trend, and prevention and control of hypertension may be one of the reasons. The standardized mortality rate of males is higher than that of females, and the mortality rate decline rate in rural areas is lower than that in urban areas. The primary and secondary prevention of AMI in such populations should be strengthened.

Objective:To analyze the trends of coronary heart disease mortality in Tengzhou city of Shandong province from 2013 to 2021.Methods:The data of coronary heart disease (ICD-10: I20-I25) from January 2013 to December 2021 were obtained from the Chronic Disease Surveillance Information System-Coronary Heart Disease Management Module of Shandong province; the population data were provided by Tengzhou Public Security Bureau. The 95% confidence interval ( CI) of the rate was calculated using the binomial exact method. The age-standardized mortality rate (ASMR) was calculated based on 2020 National Population Census. The trends of mortality rate were analyzed using the Cochran-Armitage trend test. Results:During 2013 to 2021, there were 20 667 coronary heart disease deaths in Tengzhou city, with an average of 2 296 cases per year. The male-to-female ratio was 1.09∶1, and 80.98% (16 736/20 667) of the death cases were aged 65 years or older. The crude mortality rate and ASMR of coronary heart disease were 131.84/10 5 and 168.22/10 5 in 2013, respectively. The crude mortality rate increased by 13.67% from 2013 to 2021 ( P<0.001) with an average annual increase of 1.59%; while the ASMR decreased by 18.65% from 2013 to 2021 ( P<0.05) with an average annual decease of 2.34%. The crude mortality rate of coronary heart disease in men and women showed an upward trend, with the difference in the increase for women being statistically significant ( P<0.01); while the ASMR both in men and women showed a downward trend ( P<0.001). The crude death rates from 2013 to 2021 in age groups of 65 or older for men and women decreased by 22.40% and 19.73%, respectively (both P<0.001); while the crude death rate for age groups of 44 or younger in men showed an upward trend ( P<0.05). The crude death rate among urban residents decreased by 5.22% from 2013 to 2021 with an annual decrease of 0.67% ( P>0.05); while the crude death rate among rural residents increased by 19.29% with an annual increase of 2.18% ( P<0.001). The overall crude death rate among urban residents was higher than that among rural residents (all P<0.05). Conclusions:From 2013 to 2021, the crude mortality rate of coronary heart disease in Tengzhou city of Shandong province shows an increasing trend while the age-standardized mortality rate shows a decreasing trend. The crude mortality rate of coronary heart disease in people aged 65 and above shows a decreasing trend. In addition, the change trends are different between women and men, and between urban and rural areas.

A case of primary amoebic meningoencephalitis (PAM) treated in the First Affiliated Hospital of Xinxiang Medical University in March 6, 2022 was reported.The proband was a 14-year-old boy, who was admitted to the hospital because of " fever, headache and vomiting for 2 days" . Metagenomic next-generation sequencing (mNGS) was consequently conducted to examine the pathogens in cerebrospinal fluid.Naegleria infection was detected, so the child was diagnosed with PAM.The disease developed rapidly, and the patient died 29 hours after admission.In the paper, a total of 13 studies were reviewed, and 15 children with PAM were reported.Of the reported cases, only 1 case survived, 14 cases died.PAM had a low incidence, a dangerous condition, and high mortality.Most cases were diagnosed by autopsy or pathogen diagnosis in cerebrospinal fluid.This case is confirmed by mNGS of pathogens, and it is rarely reported at home and abroad.

Bezlotoxumab is a human monoclonal antibody that can bind to C. difficile toxin B and neutralize its effects. In October 2016, bezlotoxumab was approved by the food and drug administration(FDA) to reduce the recurrence of Clostridium difficile infection (CDI) in the patients aged equal or above 18 years who are receiving antibacterial therapy. This paper introduced the pharmacology, pharmacokinetics,clinical studies,adverse reactions,interactions and medication attentions of bezlotoxumab.