Objective:This study aims to explore the prevalence of hepatitis E virus (HEV) infection in patients and the screening value of serological indicators for HEV infection patients.Methods:Retrospective analysis was conducted on 97 440 cases of anti-HEV IgM and IgG simultaneously tested in two Beijing hospitals from January 1, 2018 to August 31, 2023. Among them, there were 61 005 males and 36 435 females, with an average age of 51.65±13.05 years old. According to the positivity of anti HEV specific antibodies, they were divided into anti-HEV IgM positive group (3 588 cases), anti-HEV IgG positive group (18 083 cases), and anti-HEV antibody negative group (78 892 cases). Results of HEV RNA, liver function, AFP, PIVKA-Ⅱ and PT were collected, and their basic clinical information were recorded. The prevalence of HEV infection in patients, as well as the relationship between the positivity of anti-HEV specific antibodies and the patient′s age group, HEV RNA, and clinical characteristics were analyzed.Results:Among 97 440 patients who tested anti-HEV IgM and IgG simultaneously, the positivity rate of anti-HEV IgM was 3.68% (3 588/97 440), and was 18.56% for anti-HEV IgG (18 083/97 440). The overall positivity rates of anti-HEV IgM in two Beijing hospitals from 2018 to 2023 were 2.51%, 2.53%, 3.02%, 4.59%, 5.72%, and 4.26% ( χ2=1 401.73, P<0.001), while the positivity rates of anti-HEV IgG were 12.56%, 12.32%, 12.85%, 22.65%, 27.42%, and 26.66% ( χ2=1 058.29, P<0.001). These rates showed a gradual increase until 2023 when a decline was observed. The positivity rates of anti-HEV IgM (2.28%, 3.60%, 4.47%) ( χ2=89.62, P<0.001) and IgG (4.71%, 17.86%, 25.94%) ( χ2=2 017.32, P<0.001) increased with age in patients who aged 1-30, >30-60, and over 60 years old. The age and ALB values of patients in the anti-HEV IgM positive group were lower than the IgG-positive group, while the proportion of males, TBIL, ALT, AFP and PT values were higher than the IgG-positive group, and the differences were statistically significance ( P<0.05). Furthermore, patients in both the anti-HEV IgM and IgG positive groups had higher age, male proportion, TBIL, ALT, AFP, PIVKA-Ⅱ, and PT values than the anti-HEV negative group. Additionally, both groups had lower ALB values than the anti-HEV negative group, all of which were statistically significant ( P<0.05). 2 162 HEV infected patients were grouped based on HEV RNA positivity. The proportion of anti-HEV IgM single positive, IgG single positive, IgM+IgG double positive, and antibody negative patients in the HEV RNA positive group were 5.42% (18/332), 3.62% (12/332), 90.36% (300/332), and 0.60% (2/332), respectively. Among them, the proportion of anti-HEV IgM+IgG double positive patients in the HEV RNA positive group was higher than that in the HEV RNA negative group ( χ2=302.87, P<0.001), while the proportion of anti-HEV IgG single positive ( χ2=174.36, P<0.001) and anti-HEV antibody negative patients ( χ2=59.28, P<0.001) were lower than that in the HEV RNA negative group, both of which were statistically significant ( P<0.001). In addition, the positive rates of HEV RNA in anti-HEV IgM positive, IgG positive, and antibody negative patients were 29.23% (318/1 088), 17.59% (312/1 774), and 0.65% (2/306), respectively. Conclusion:The HEV infection rate among patients declined in 2023. HEV infection is age-related, with older individuals being more susceptible. Abnormal liver function and jaundice were commonly observed during HEV infection. It is crucial to note that the absence of anti-HEV specific antibodies cannot rule out HEV infection; therefore, additional testing for HEV RNA and/or HEV Ag is necessary for accurate diagnosis.

Objective:To investigate the tissue methylation features of adenocarcinoma patients presenting as pulmonary nodules on CT scans.Methods:A retrospective analysis was conducted on 70 adenocarcinoma patients with pulmonary nodules diagnosed at the Shanghai General Hospital from June 1, 2022 to January 20, 2024. Participants were assigned to two groups using the random number table, with 40 in the discovery group and 30 in the validation group. In the discovery group, tissue samples were analyzed using reduced representation bisulfite sequencing (RRBS) technology to compare the average methylation levels between cancer tissues and paired adjacent non-cancerous tissues. Differentially methylated regions (DMRs) were screened for analysis of their distribution across various genomic functional elements, and hierarchical clustering was plotted. GO and KEGG pathway enrichment analyses were further conducted on the DMRs. Subsequently, candidate DMRs associated with lung adenocarcinoma were validated using TCGA lung adenocarcinoma cohort and targeted bisulfite sequencing technology in the validation group. The comparison of methylation levels between groups was conducted using t-tests or non-parametric tests, while rates and composition ratios were analyzed using chi-square tests or Fisher′s exact test.Results:In discovery cohort, the average methylation level in cancer tissues was lower compared to adjacent normal tissues [(42.369±4.627) vs (44.370±4.046), t=?2.059, P=0.043]. A total of 37 995 DMRs were identified, including 16 889 upregulated regions and 21 106 downregulated regions, predominantly locating in promoter regions (48.917%), introns (36.457%), and exons (10.812%). The DMR clustering heatmap revealed two distinct clusters corresponding to cancer tissues and adjacent non-cancerous tissues. GO analysis showed that DMRs associated genes were mainly located in the cell membrane and nuclear chromatin, and were primarily involved in RNA polymerase Ⅱ-related transcription and regulation. KEGG pathway enrichment analysis indicated that DMRs associated genes were mainly involved in neuroactive ligand-receptor interaction, cancer pathways, calcium signaling pathway, cAMP signaling pathway, and MAPK signaling pathway. Validation in the TCGA cohort confirmed 11 potential characteristic DMRs. In the validation group, TBS confirmed that the methylation levels of DMRs associated with MIR10B, DMRTA2, HOPX, TFAP2B and MARCH11 in cancer tissues were significantly higher than those in adjacent non-cancerous tissues [11.200(4.305, 27.088) vs 2.650(1.298, 4.645), Z=?4.539, P<0.05; 18.610(13.600, 33.025) vs 8.675(5.488, 13.085), Z=?4.554, P<0.05; 17.600(2.183, 76.015) vs 1.085(0.898, 1.835), Z=?5.131, P<0.05; 5.250(3.220, 7.693) vs 3.495(2.165, 4.383), Z=?2.861, P<0.05; 11.515(7.525, 21.033) vs 7.830(5.518, 11.488), Z=?2.440, P<0.05 ], and the differences were statistically significant. Conclusions:Lung adenocarcinoma tissue exhibits different methylation patterns compared with adjacent normal lung tissue. The identified DMRs are involved in the regulation of several key pathways. Results from the TCGA cohort and an independent validation group support the potential diagnostic value of DMRs such as MIR10B, DMRTA2, HOPX, TFAP2B, and MARCH11 in lung adenocarcinoma, though their clinical application requires further validation.

Objective:To evaluate the effect of dexmedetomidine on the extracellular signal-regulated kinase(ERK)/sodium-potassium ATPase(Na + -K + -ATPase)signaing pathway in lung tissues of rats with mechanical ventilation-induced lung injury (VILI). Methods:Forty-eighty clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 4 groups ( n=12 each) using a random number table method: control group (group C), VILI (alpha2-adrenergic receptor antagonist) group (group V), dexmedetomidine group (group D), and dexmedetomidine plus yohimbine group (group DY). Group C underwent no mechanical ventilation and breathed air spontaneously for 4 h. Mechanical ventilation (respiratory rate 40 breaths/min, tidal volume 40 ml/kg, inspiratory/expiratory ratio 1∶1, PEEP 0, fraction of inspired oxygen 21%) lasted 4 h in group V. Dexmedetomidine was infused intravenously in a dose of 5.0 μg/kg at 20 min before ventilation followed by an infusion of 5.0 μg·kg -1· h -1 throughout ventilation in group D. In group DY, yohimbine 0.1 mg/kg was injected intravenously at 10 min before dexmedetomidine, and the other treatments were similar to these previously described in group D. Blood samples and lung tissues were taken at 4 h of mechanical ventilation to determine the wet/dry weight ratio (W/D ratio), lung permeability index (LPI), alveolar fluid clearance rate (AFC), and expression of extracellular signal-regulated kinase (ERK), phosphorylated extracellular signal-regulated kinase (p-ERK), and Na + -K + -ATPase in lung tissues (by Western blot) and to observe pathological changes of lung tissues. Results:Compared with group C, LPI and W/D ratio were significantly increased, AFC was decreased, p-ERK expression was up-regulated, and Na + -K + -ATPase expression was down-regulated in group V and group DY ( P<0.05), and no significant change was found in the incidence of the parameters mentioned above in group D ( P>0.05). Compared with group V, LPI and W/D ratio were significantly decreased, AFC was increased, p-ERK expression was down-regulated, Na + -K + -ATPase expression was up-regulated ( P<0.05), and the pathological changes of lung tissues were significantly attenuated in group D, and no significant change was found in the incidence of the parameters mentioned above in group DY ( P>0.05). Compared with group D, LPI and W/D ratio were significantly increased, AFC was decreased, p-ERK expression was up-regulated, Na + -K + -ATPase expression was down-regulated ( P<0.05), and the pathological changes of lung tissues were accentuated in group DY. Conclusion:The mechanism by which dexmedetomidine alleviates VILI may be related to activating alpha2-adrenergic receptors and inhibiting ERK/Na + -K + -ATPase signaling pathway in rats.

Objective To investigate the effect of rhynchophylline on mRNA expression of excitatory amino acid transporter 2 (EAAT2 ) and N-methyl-D-aspartic acid receptor 2B (NR2B) after astrocyte oxygen-glucose deprivation. Methods The subcultured third generation astrocytes from the hippocampus were inoculated into 6-well plates, and they were divided into blank control group, hypoxia-ischemia group,low-dose rhynchophylline group (0. 02 mg/ml) and high-dose rhynchophylline group (0. 2 mg/ml) after the cells were attached to the wall and grew out protrusion. The total RNAs in each group were extracted.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of EAAT2 and NR2B mRNA in astrocytes of each group. Results Compared with the blank control group, the expression levels of NR2B and EAAT2 mRNA in astrocytes of the ischemia-hypoxia group were significantly higher (all P < 0. 05 ). The expression levels of NR2B and EAAT2 mRNA in the low-dose rhynchophylline group were lower than those in ischemia-hypoxia group, but there was no significant difference. The expression levels of NR2B and EAAT2 mRNA in the high-dose rhynchophylline group were significantly lower than the ischemia-hypoxia group and the low-dose rhynchophylline group (all P < 0. 05).Conclusion The expression of EAAT2 and NR2B mRNA in astrocytes of hippocampus cultured in vitro was significantly increased after ischemia and hypoxia, and rhynchophylline intervention could significantly reduce its expression in a concentration dependent manner.

AIM:To investigate the protective effects of astrocyte protein phosphatase 2A (PP2A) up-regula-tion on APP/PS1 double transgenic mice.METHODS:An eGFP-wtPP2A lentivirus with glial fiber acidic protein promoter was constructed to specifically increase PP2A expression in the astrocytes.The mice were divided into wild -type mice +vector virus group (Con), APP/PS1 transgenic mice +vector virus group (APP/PS1) and APP/PS1 transgenic mice +eGFP-wtPP2A lentivirus group (PP2A) by lateral ventricular injection of the lentivirus.Four weeks after injection of the vi-rus, the immunofluorescence of brain slices were used to detect the level of β-amyloid protein ( Aβ) .Golgi staining was used to detect the changes of dendritic spine density and morphology.Electron microscopy was applied to detect the thickness of postsynaptic density (PSD).The Morris water maze test was applied to examine the learning and memory abilities of the mice.RESULTS: Up-regulation of PP2A in the astrocytes attenuated Aβlevel increasing in APP/PS1 group.Up-regulation of PP2A in the astrocytes significantly attenuated both decreases in the dendritic spine density and the percentage of mushroom-like dendritic spines in the hippocampal CA3 region of APP/PS1 mice.Up-regulation of PP2A in the astrocytes significantly attenuated the reduced thickness of PSD in APP/PS1 group.Up-regulation of PP2A in the astro-cytes attenuated the escape latency extending in APP/PS1 group .CONCLUSION: Up-regulation of PP2A in the astro-cytes reduces AD-like pathological changes, and attenuates synaptic impairment, synaptic plasticity deficits and cognitive impairment in the APP/PS1 double transgenic mice.

Objective To estimate the diagnostic value of circulating tumor cell detection for non-small cell lung cancer.Methods A Non-intervention clinical study was conducted in this research.From October 2014 to April 2015, totally 162 NSCLC who presented at Thoracic Surgery Department, 119 benign pulmonary disease and 52 healthy individuals were collected from Shanghai Chest Hospital.Folate receptor ( FR) based polymerase chain reaction ( PCR) method was used to detect the circulating tumor cell ( CTC) level, CEA and CYFRA21-1 was detected by the flowcytometry fluorescence luminance method, SCC was detected with Chemiluminescent microparticle immunoassay.The differences among groups were analyzed by the Kruskal-Wallis test( multi group comparison) and the Mann-Whitney U test( two group comparison) , and the chi-square test was used in the positive rate comparison;the Receiver Operating Characteristics ( ROC) curve was established.Results The median level of CTC in NSCLC patients was 11.90 Units/3 ml, which was significantly higher than those of benign pulmonary disease ( 6.72 CTC Units/3 ml ) and healthy individuals (5.82 CTC Units/3 ml,χ2 =125.990, P<0.01).Areas Under Curve ( AUCs) of ROC curve for NSCLC was 0.853 2(95% CI: 0.809 5,0.896 9).The cut-off value for discriminating NSCLC with benign pulmonary disease/healthy people was 8.74 CTC Units/3 ml with sensitivity being 77.16% and specificity being 90.06%.The positive rate of CTC in Stage I NSCLC patients was 68.7%, which was much higher than that of the combination of tumor markers(χ2 =32.98,P<0.01).Conclusion With relatively high sensitivity and specificity, the detection of circulating tumor cell may has a clinical value of application and extension.

Objective To evaluate the application value of circulating tumor cell ( CTC ) in the differential diagnosis of solitary pulmonary nodule ( SPN ) . Methods Peripheral blood samples were collected from 134 patients with solitary pulmonary nodule in Shanghai Chest Hospital from September 2013 to January 2015, including 80 patients with malignant nodule and 54 with benign nodule.CTC levels of the above subjects were detected by ligand-targeted polymerase chain reaction ( LT-PCR ) assay, and serum carcinoembryonic antigen ( CEA ) and cytokeratin 19 fragment ( CYFRA21-1 ) were detected by flow fluorescence assay.Results By Mann-Whitney U Test, the CTC levels of malignant SPN patients [11.06 (8.77-14.41)units/3 ml] were significantly higher than those of benign SPN patients [6.65(4.49 -7.84)units/3 ml] (Z=-6.217,P<0.001).The sensitivity and specificity of differential diagnosis of SPN for CTC were 80%(64/80) and 85%(46/54) respectively.According to the diameter of SPN, the patients were divided into three groups to evaluate the diagnostic value of CTC in SPN with different size .For SPN with diameter less than 8 mm, the sensitivity and specificity of CTC were 6/9 and 4/5 respectively .For SPN with diameter between 8 mm and 20 mm, the sensitivity and specificity of CTC were 83%(35/42) and 85%(29/34).For SPN with diameter greater than 20 mm, the sensitivity and specificity of CTC were 79%(23/29) and 13/15.Conclusion Comparing with the traditional tumor markers, CTC could provide more clinical value in the differential diagnosis of solitary pulmonary nodule .

Objective]Through judging the constitutional types of traditional Chinese medicine(TCM) of acute mastitis patients, do statistics and analysis of the distribution of the acute mastitis patients’constitutional types of TCM, and study their laws, in order to provide guiding basis of constitutional theory of TCM for preventing and early treating mastitis. [Method] By means of questionnaire method, judge the constitutional types of TCM of 77 acute mastitis patients according to the classification and judgment of constitution of TCM ,and analyze the results by statistical methods. [Results] The number of the constitutional types of TCM of 77 acute mastitis patients from high to low is as follows: dampness-heat constitution is 65 cases, energy-stagnant constitution is 52 cases, yang-deficiency constitution is 51 cases, qi-deficiency constitution is 37 cases, yin-deficiency constitution is 34 cases, phlegm-dampness constitution is 32 cases, blood-stasis constitution is 29 cases, normal constitution is 12 cases, and special-natural constitution is 7 cases. Individuality constitution has 9 cases, two and three mixed constitution each has 12 cases, four and five mixed constitution each has 9 cases, six mixed constitution has 14 cases, seven mixed constitution has 11 cases, eight mixed constitution has 1 case. In the main constitutional types of TCM of 77 acute mastitis patients, dampness-heat constitution(54 cases, 70.13%) is most, followed by normal constitution(12 cases, 15.58%) and yang-deficiency constitution(5 cases, 6.49%). In the individuality constitution and two mixed constitution, the proportion of dampness-heat constitution(60 cases, 39.47%) is highest, and that of yang-deficiency constitution(34 cases, 22.37%) is also higher. This shows that: 1. Dampness-heat constitution is the most common constitutional type related with the occurrence of acute mastitis, and yang-deficiency constitution is the second. 2. The acute mastitis patients have a variety of constitutional types such as dampness-heat constitution, energy-stagnant constitution etc. Mixed constitution is common and complex. May be it is associated with the particular period of postpartum lactation period. [Conclusion] Constitution as internal factors of disease has an important influence on occurrence and development of acute mastitis. It has great meaning to adjust prejudiced constitutional types of TCM as dampness-heat constitution and yang-deficiency constitution for preventing and early treating acute mastitis. This reflects the TCM thought of“treat unoccurred disease”.

Objective To investigate the clinical significance of serum anti-Saccharomyces cerevisias antibody (ASCA),anti-outer membrane porin C (anti-OmpC),antibody to Pseudomonas fluorescens-associated sequence I2 (anti-I2 )and antibody to bacterial flagellin (anti-CBirl )in the diagnosis and treatment of inflammatory bowel disease (IBD).Methods From 2011 to 2013,87 patients with IBD were enrolled and divided into Crohn′s disease (CD)group (66 cases)and ulcerative colitis (UC)group (21 cases).A total of 62 age and gender matched healthy individuals were enrolled as the control group. Fasting blood samples (2 mL)of the subjects were collected.The expression of ASCA,anti-OmpC,anti-I2 and anti-Cbirl antibodies was detected with enzyme-linked immunosorbent assay (ELISA)kits.The diagnosis value of each antibody in IBD and the differential diagnostic value of in UC and CD were compared by receiver operating characteristic (ROC)curve.Results The area under the curve (AUC)of ASCA between IBD and the healthy control group,between CD group and UC group was 0.580 and 0.512, respectively;the accuracy in diagnosis was low.The AUC of anti-CBirl between IBD and the healthy control group was 0.617.There was no differential diagnosis significance of the other antibodies.The positive rate of ASCA in IBD group was 62.1 % (54/87),which was significantly higher than that in the control group (38.7%,24/62).The positive rates of anti-OmpC and anti-I2 in IBD group was significantly lower than those in the control group and the differences were statistically significant (both P <0.05).No difference was observed in positive rates of serum antibodies among the others groups (all P >0.05).The specificity,sensitivity,positive predictive value (PPV)and negative predictive value (NPV)of ASCA in differential diagnosis of CD and UC was 52.4%,66.7%,81 .48% and 33.33%,respectively.The specificity and sensitivity of anti-OmpC,anti-I2 and anti-CBirl in differential diagnosis of CD and UC was 81 .0% to 100.0% and 9.1 % to 37.9%,respectively.The specificity,sensitivity,PPV and NPV of double-positive ASCA and anti-I2 in the diagnosis of CD was 57.1 %,86.4%,82.6% and 50.0%, respectively.The positive rate of ASCA and anti-I2 in CD group was significantly higher than that in UC group (84.8%(56/66)vs 57.1 % (12/21 );χ2 =5 .633,P =0.018 ).Conclusions Positive ASCA has some significance in the diagnosis of patients with IBD in our country.The detection of anti-I2 can help to diagnose ASCA negative CD.Because of low sensitivity and positive rate,anti-OmpC and anti-CBirl have limited value in the diagnosis of IBD and the differential diagnosis of UC and CD in our country.

Objective To explore the findings of diaphysial osteosarcoma in long bone on X-ray,CT and MRI, and discuss their clinical features and manifestations for differential diagnosis. Methods Twenty-eight cases with diaphysial osteosarcoma in long bone proved by surgery and pathology were reviewed retrospectively. Eighteen tumors were located in the femur, 4 in fibula, 4 in humerus and 2 in tibia. All of the patients were examined by X-ray, CT and MRI. The imaging manifestations on X-ray, CT and MRI were analyzed, and the relationship of the imaging features with the pathological types was also observed. The imaging signs were correlated with the pathologic findings with a double blind method. Results Of the 28 cases, there were 16 cases with large bone destruction, 22 cases with periosteal reaction on X-ray and CT. On X-ray, 18 cases showed soft tissue mass and 12 cases with neoplastic bone and tumor calcification.While on CT, 22 cases showed soft tissue mass on plain scan and 2 more cases displayed soft tissue mass after the injection of contrast mediun. Sixteen cases showed neoplastic bone and tumor calcification on CT.On MRI, there were 10 cases with bone destruction and periosteal reaction with iso- and hypo-intense on T1WI and iso- signals on T2WI. Twenty-six cases showed soft tissue edema and bone marrow on MRI. The soft mass were iso-signals on T1 WI and iso-hyperintense signals on T2 WI or STIR. The soft tissue edema was found hyperintense signals on T2WI or STIR. The lesions had heterogeneous enhancement especially in bone marrow with edema and adjcent soft tissue. Conclusion The X-ray, CT and MRI can reflect the pathological changes of diaphysial osteosarcoma in long bone from different aspects. Lower incidence, large bone destruction and no pathological fracture were the features of diaphysial osteosarcoma. The osteogenic type is diagnosed easily, but the osteolytic lesion should be differentiated from Ewing sarcoma, malignant giant cell tumor of bone and so on.