Objective To establish a rat model of acute antibody-mediated rejection (AMR) in kidney transplantation and investigate the preventive effect of dihydroartemisinin (DHA) on acute AMR. Methods BN rats were used as donors and Lewis rats as recipients. Kidney transplantation was performed 2 weeks after skin transplantation for sensitization. After establishing the acute AMR model in rat kidney transplantation, the recipients of experimental groups included the syngeneic kidney transplantation group (6 rats), the allogeneic kidney transplantation group (6 rats), the syngeneic skin transplantation followed by kidney transplantation group (12 rats), and the allogeneic skin transplantation followed by kidney transplantation group (24 rats). The groups for investigating the preventive effect of DHA on acute AMR included the control group (allogeneic skin transplantation followed by kidney transplantation) and the DHA group (allogeneic skin transplantation followed by kidney transplantation + DHA), with 12 rats in each group. The survival time of recipient rats, serum donor-specific antibody (DSA) levels and graft pathological changes were used to identify the acute AMR model. On this basis, DSA levels, pathological changes in the transplant kidneys and peripheral blood B-cell levels were detected to assess the preventive effect of DHA on acute AMR. Results Compared with the allogeneic kidney transplantation group, skin transplantation sensitization significantly shortened the survival time of recipient rats (P<0.01). Compared with the syngeneic skin transplantation followed by kidney transplantation group, the allogeneic skin transplantation followed by kidney transplantation group showed significantly elevated serum DSA-IgG levels from 7 days after skin transplantation to 5 days after kidney transplantation (P<0.01), and significantly elevated DSA-IgM levels at 7 and 14 days after skin transplantation(all P<0.01). The transplant kidneys in the allogeneic skin transplantation followed by kidney transplantation group showed a small number of inflammatory cell infiltrations, tubular necrosis, capillaritis, and C4d deposition starting from 1 day after kidney transplantation, with these pathological changes worsening as the post-transplantation days increased. The kidney damage became significant starting from 3 days after transplantation. The above pathology manifestations were consistent with the characteristics of acute AMR. On the basis of establishing the acute AMR model, DHA treatment significantly prolonged the survival time of recipient rats (P<0.01) , and reduced serum DSA-IgG and DSA-IgM levels. DHA treatment significantly alleviated the pathological manifestations of acute AMR, including kidney damage, inflammatory cell infiltration, capillaritis and tubular necrosis, and also reduced C4d deposition in the transplant kidneys, inflammatory cell infiltration and peripheral blood CD19⁺ B-cell levels. Conclusions An acute AMR model is established by performing kidney transplantation 2 weeks after allogeneic skin transplantation in rats. It is discovered that DHA has immunosuppressive effects and may effectively prevent acute AMR, which provides a new strategy for the management of clinical AMR.

BACKGROUND:Oxidative injury is considered to be one of the important factors of cerebral ischemia-reperfusion injury.Superoxide dismutase 2(SOD2)is a key mitochondrial antioxidant molecule,and fenofibrate can regulate the expression of SOD2 by activating peroxisome proliferator-activated receptor α. OBJECTIVE:To explore whether the mechanism of fenofibrate in the treatment of cerebral ischemia-reperfusion injury depends on the expression of SOD2. METHODS:The TALENs system was used to construct SOD2 transgenic mice.The transgenic mice were genotyped by PCR and DNA sequencing techniques.The expression of SOD2 protein in transgenic mice was detected by western blot assay.Wild-type and SOD2 transgenic mice were randomly divided into four groups:wild-type control group(n=6),wild-type fenofibrate group(n=6),SOD2 transgenic control group(n=5)and SOD2 transgenic fenofibrate group(n=5).A mouse model of middle cerebral artery occlusion was prepared using the suture-occlusion method.After 90 minutes of ischemia,the thread was removed to reperfuse cerebral blood flow for 30 minutes.A cerebral blood flow monitor was used to monitor local cerebral blood flow.Brain tissue slices were taken for 2,3,5-triphenyltetrazolium chloride staining to analyze the situation of cerebral infarction in each group. RESULTS AND CONCLUSION:After PCR and DNA sequencing analysis,nine SOD2+/+ transgenic mice were successfully constructed.After cerebral ischemia-reperfusion,the wild-type fenofibrate group showed partial recovery of cerebral blood flow and significantly reduced cerebral infarction volume compared with the wild-type control group(P<0.001).There was no significant difference in cerebral blood flow and cerebral infarction volume between the SOD2 transgenic fenofibrate group and the SOD2 transgenic control group.The SOD2 transgenic control was superior to the wild-type control group in terms of improving cerebral blood flow and cerebral infarction(P<0.001).There were also no significant differences in cerebral blood flow and cerebral infarction volume between the wild-type fenofibrate group and the SOD2 transgenic control group and between the wild-type fenofibrate group and the SOD2 transgenic fenofibrate group.To conclude,the expression of SOD2 is one of the mechanisms of fenofibrate in the treatment of cerebral ischemia-reperfusion injury.

OBJECTIVES: To investigate the clinical characteristics and risk factors for severe events of coronavirus disease 2019 (COVID-19) in elderly patients. METHODS: Retrospective analysis was performed on the clinical data of all elderly COVID- 19 patients treated in Changsha Public Health Treatment Center from January 17, 2020 to March 15, 2020, which included basic diseases, symptoms, test results, and other clinical characteristics, and prognostic indicators such as severity of illness, length of hospital stay, virus shedding time and mortality rate. The differences in clinical characteristics and prognostic indicators between elderly, middle-aged, and young COVID-19 patients were also analyzed. Logistic regression model was used to conduct univariate and multivariate analysis of risk factors for developing severe events in elderly COVID-19 patients; receiver operating characteristic (ROC) curve analysis was used to evaluate the prediction efficacy. RESULTS: Of the 230 COVID-19 adult patients, 34 were young patients (14.8%), 136 were middle-aged patients (59.1%), and 60 were elderly (26.1%). Among the 60 elderly patients, 23 were male (38.3%) and 37 were female (61.7%), with a medium age of 66 years old. Common symptoms were fever (66.7%), cough (50.0%), and fatigue (41.7%). C reactive protein (CRP) was increased significantly. The proportion of severe cases was 31.7%, and mortality was 1.7%. The median length of hospitalization and median virus shedding time were 18.5 days and 21 days, respectively. Compared with the young and the middle-aged patients, the elderly had a higher proportion of hypertension, diabetes, and cardiovascular diseases, more common shortness of breath, higher proportions of pneumonia and severe cases (all <0.05), and the decreased lymphocyte count and lymphocyte percentage (both <0.05), as well as higher CRP and erythrocyte sedimentation rate (ESR) levels (both <0.05). Compared with non-severe cases, severe elderly patients demonstrated higher CRP and aspartate aminotransferase (AST) levels (all <0.05), the reduced lymphocyte count (<0.05), and the prolonged length of hospitalization and virus shedding duration (both <0.05). Univariate logistic regression analysis indicated that the lymphocytes proportion, CRP and AST levels were significantly correlated with the risk for developing severe events in elderly COVID-19 patients (all <0.05). Multivariate logistic regression found that severe events in elderly patients with COVID-19 were significantly correlated with CRP level (OR=1.041, =0.013). ROC curve analysis revealed that the area under the curve (AUC) for CRP to diagnose severe events in elderly COVID 19 patients was 0.851. CONCLUSIONS The proportion of severe cases in elderly COVID-19 patients is higher than that in young and middle-aged patients. CRP level has a good predictive value for the possibility of severe events in elderly COVID-19 patients.
Adult ; Aged ; Betacoronavirus ; C-Reactive Protein ; analysis ; China ; Comorbidity ; Coronavirus Infections ; diagnosis ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnosis ; physiopathology ; Retrospective Studies ; Risk Factors

Brucellosis is a zoonotic,acute and chronic systemic disease caused by Brucella,the incidence is increasing year by year.Brucella can invade many organs and tissues of human body,affect working ability,and even cause disability and death,seriously endanger human health.Now the main treatment method of brucellosis is multi-course and combined application of antibacterial drugs,but long-term using of such drugs is prone to drug resistance and adverse reactions.In recent years,the combination of traditional Chinese and western medicine has been applied to treat brucellosis and achieved certain curative effect.In this paper,the current situation in treating brucellosis by integrated traditional Chinese and western medicine is summarized and prospected.

Objective To investigate the associations of cognitive function with apolipoprotein E (APOE) gene polymorphism and chronic diseases among long-lived people in Zhongxiang City of Hubei Province.Methods A total of 110 long-lived elderly residents aged 90 years and over were collected.Their cognitive function was determined face-to-face using questionnaires by trained interviewers.According to mini-mental state examination(MMSE) scores,subjects were divided into a no dementia risk group and a high dementia risk group.General demographic characteristics,activities of daily living,depression state and nutrition status were compared between the two groups.Correlations of dementia with APOE gene polymorphism and chronic diseases were analyzed.Results The average MMSE score was 22.3±4.8.Among the 110 long-lived people,18 cases had a high risk for dementia,accounting for 16.4％,and 92 cases had no risk of dementia,accounting for 83.6％.The risk of dementia in long-lived elderly people was correlated with activities of daily living,mental state,nutritional status and falls(all P＜0.05).There were 8 cases with the APOE gene ε4/ε3 genotype in the high dementia risk group and 16 cases with the APOE gene ε4/e3 genotype in the no dementia risk group,with the former group showing a higher frequency of the APOE ε4/ε3 genotype (44.4％ vs.17.4％,x2 =6.46,P＜0.05).The former group also seemed to have a higher APOE ε4 frequency,but the difference was not statistically significant(22.2 ％ vs.10.3 ％,x2 =3.96,P =0.055)Chronic diseases prevalent in the long-lived people were hypertension(86 cases,78.2 ％),hearing loss (72 cases,65.5％),hyperlipidemia(56 cases,50.9％),anemia(43 cases,39.1％),impaired vision(39 cases,35.5 ％),chronic kidney diseases(25 cases,22.7 ％),chronic heart diseases (18 cases,16.4 ％) and osteoarthritis(18 cases,16.4％).No correlation was found between the risk of dementia and chronic diseases(P＞0.05).Conclusions Cognitive function is highly correlated with activities of daily living,mental state and nutritional status among long-lived elderly people in Zhongxiang City.The risk of dementia has a correlation with the APOE gene e4/e3 genotype but not with chronic diseases in long-lived people.

OBJECTIVE: To carry out phenotypic and genotypic analysis for two Chinese pedigrees affected with coagulation factor XII (F XII) deficiency. METHODS: Plasma prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), and blood coagulation factor VIII, IX, XI, XII activity (FVIII:C, FIX:C, FXI:C, FXII:C) were determined with one stage clotting assay on a STAGO coagulation analyzer. FXII antigen was determined with an enzyme linked immunosorbent assay (ELISA). The 14 exons and their flanking sequences of the F12 gene were subjected to PCR amplification and Sanger sequencing. The conservation and structure of mutant protein were analyzed with MegAlign software and PYMOL software. RESULTS: The APTT of the probands was significantly prolonged, while their FXII:C and FXII:Ag were significantly reduced. Genetic analysis of the proband has revealed three novel mutations in the F12 gene, including g.5972G>A splice site mutation in intron 5, g.8810_8814delGTCTA in exon 14, and g.6259G>A (p.Pro182Leu) in exon 7. In addition, a previously known mutation IVS13-1G>A has been found. CONCLUSION Four mutations have been identified in the two Chinese pedigrees, among which three were novel. Above mutations probably played a role in the defect of FXII in the two pedigrees.
Exons ; Factor XII ; genetics ; Factor XII Deficiency ; genetics ; Genetic Testing ; Humans ; Pedigree

OBJECTIVETo explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with FX deficiency.

METHODSProthrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen, FII activity(FII:C), FVII activity(FVII:C), FIX activity (FIX:C), FX activity(FX:C) were determined with a one-stage clotting assay. The FX antigen(FX:Ag) was detected with an enzyme linked immunosorbent assay(ELISA). The 8 exons, introns and 5' and 3' untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing. Suspected mutation was confirmed by reverse sequencing. Polymorphisms were excluded by direct sequencing of 100 healthy individuals.

RESULTSThe PT and APTT of the proband have prolonged to 16.1 s and 49.0 s, respectively. Her FX:C and FX:Ag were reduced by 27% and 56%, and her mother's PT, APTT, FX:C and FX:Ag were 14.8 s, 37.4 s, 44%, 34%, respectively. Her grandmother's PT, APTT, FX:C and FX:Ag were 15.8 s, 42.2 s, 31%, 45%, respectively. The results of her father and other family members were all within the normal range. Genetic analysis has revealed a heterozygous G to A mutation in the proband at position 28076 in exon 8 of the F10 gene, which resulted in a p.Gly363Ser substitution. The same mutation was also found in her mother and grandmother. No mutation of the F10 gene was found in her father. Gly363Ser may result in changes in the secondary structure of the FX protein and reduction of its activity.

CONCLUSIONThe g.28076G to A(p.Gly363Ser) mutation of the F10 gene probably underlies the FX deficiency in this pedigree. The mutation was discovered for the first time in Chinese patients.

Objective To observe the effects of massage on inflammation,oxidative stress and autophagy during the repair of acute contusion of skeletal muscles so as to explore its biological mechanisms.Methods Forty-two adult Sprague-Dawley rats were randomly divided into a control group (n =6),a model group (n =18),and a treatment group (n =18).Acute contusion of the gastrocnemius muscles of the rats in the model and treatment groups was inflicted using a home-made impactor.Beginning forty-eight hours later,15 minutes of massage was administered daily for two weeks.After one,7 and 14 days of the massage treatment,the injured gastrocnemius was resected from 6 rats of both the model and treatment groups.Morphological changes were observed using haematoxylin and eosin (HE) staining.The serum content of tumor necrosis factor alpha (TNF-α),interleukin1β (IL-1 β),C reactive protein (CRP) and prostaglandin E2 (PGE2) were detected using enzyme-linked immunosorbent assay (ELISA).The serum content of superoxide (SOD) and malondialdehyde (MDA) were detected using spectrophotometry.The expression of microtubule-associated protein 1 light chain 3 (LC3),Bcl-2 homeodomain protein Beclin1 and ubiquitin binding protein P62 were detected using Western blotting.Results The HE staining showed more significant collapse and swelling of cells in the model group than in the control group at each time point.New muscle cells were observed at days 7 and 14 in the model group.At each time point,significantly better recovery was observed in the treatment group compared to the model group,with more new muscle cells and better cell morphology.According to the ELISA results,a significant increase in serum pro-inflammatory factors occurred in the model group compared to the control group and compared to the treatment group after one day and 7 days of treatment.The average serum content of SOD and MDA in the model group was significantly higher than in the control group,while the average serum content of SOD in the treatment group was significantly higher than in the model group and that of MDA was significantly lower.Western blotting showed a significant decrease in LC3 (Ⅱ/Ⅰ) and Beclin1,as well as a significant increase in P62 in the model group at each time point compared with the treatment group and the controls.Conclusion Inflammation and oxidative stress increase significantly in a skeletal muscle after injury,but autophagy decreases significantly.Massage can effectively reduce the inflammatory response and oxidative stress and promote autophagy,which leads to quicker repair of skeletal muscles.

Objective To study the advantage of using axillary fascia swelling in cleaning the lymph nodes in modified radical mastectomy. Methods 866 patients with breast cancer collected from June 2005 to June 2011 were divided into the observation group (578 cases, axilla-ry fascia swelling method) and the control group (288 cases, standard operation method), and the postoperative complications of the two groups were compared. Results In the observation group, there were 8 cases (1. 4%) of upper limb lymphatic edema, 13 cases (2. 2%) of chest wall and upper limb paresthesia, and 8 cases (1. 4%) of lymphatic leakage. The complications of the observation group were significantly reduced compared with the control group (P<0. 05), and there was no increase in the local recurrence rate (P<0. 05). Conclusion Axillary fascia swell-ing applied in cleaning lymph nodes could obviously reduce the postoperative complications after modified radical mastectomy.

Objective To evaluate the application value of ultrasound-guided Mammotome minimally invasive surgery system in non-palpable breast lesions.Methods A total of 325 lesions of 245 patients with breast lumps by ultrasound examination were under ultrasound-guided Mammotome minimally invasive surgery system.Results Accurate and complete resection of all of the 325 breast lumps was achieved,5 lesions with hematoma,the incidence rate was 1.54％ (5/325).The pathologic diagnoses included:fibro-adenoma (172 lesions),intraductal papilloma (8 lesions),fibroadenosis (87 lesions),fibro-cystic adenosis (56 lesions),invasive ductal carcinoma (2 lesions).Conclusion The application of ultrasound-guided Mammotome minimally invasive surgery system in non-palpable beast lesions has advantages of accurate positioning,minimally invasive,beautiful.