Objective:The transjugular or transfemoral approach is used as a common method for hepatic venous pressure gradient (HVPG) measurement in current practice. This study aims to confirm the safety and effectiveness of measuring HVPG via the forearm venous approach.Methods:Prospective recruitment was conducted for patients with cirrhosis who underwent HVPG measurement via the forearm venous approach at six hospitals in China and Japan from September 2020 to December 2020. Patients' clinical baseline information and HVPG measurement data were collected. The right median cubital vein or basilic vein approach for all enrolled patients was selected. The HVPG standard process was used to measure pressure. Research data were analyzed using SPSS 22.0 statistical software. Quantitative data were used to represent medians (interquartile ranges), while qualitative data were used to represent frequency and rates. The correlation between two sets of data was analyzed using Pearson correlation analysis.Results:A total of 43 cases were enrolled in this study. Of these, 41 (95.3%) successfully underwent HVPG measurement via the forearm venous approach. None of the patients had any serious complications. The median operation time for HVPG detection via forearm vein was 18.0 minutes (12.3~38.8 minutes). This study confirmed that HVPG was positively closely related to Child-Pugh score ( r = 0.47, P = 0.002), albumin-bilirubin score ( r = 0.37, P = 0.001), Lok index ( r = 0.36, P = 0.02), liver stiffness ( r = 0.58, P = 0.01), and spleen stiffness ( r = 0.77, P = 0.01), while negatively correlated with albumin ( r = -0.42, P = 0.006). Conclusion:The results of this multi-centre retrospective study suggest that HVPG measurement via the forearm venous approach is safe and feasible.

Objective:To investigate the protective effect of racanisodamine (654-2) on lung epithelial cell injury induced by X-ray in mice and unravel the underlying mechanism.Methods:Mouse alveolar epithelial cells MLE-12 were used to establish radiation-induced lung injury (RILI) model in vitro and divided into 4 groups as follows: control (no irradiation), radiation (16 Gy radiation), treatment 1 (16 Gy radiation + 2 μmol/L 654-2), treatment 2 (16 Gy radiation + 10 μmol/L 654-2), and inhibitor (16 Gy radiation + 10 μmol/L 654-2 + 2 μmol/L ML385), respectively. Cells were sampled at different time points after radiation. Cell senescence was detected with senescence-associated β-galactosidase (SA-β-Gal) staining. Cell colony-forming ability was detected to observe the recovery capability of cells after treatment. The expression levels of p21, p16, phosphorylated histone H2AX(γH2AX), nuclear factor erythroid-2 related factor 2 (Nrf2), Nrf2 Ser40 site phosphorylation (p-Nrf2), p62, heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1) proteins were measured by Western blot. Cell apoptosis and the production of intracellular reactive oxygen species (ROS) were determined by flow cytometry. The expression levels of glutathione (GSH) and superoxide dismutase (SOD) were detected according to the manufectuer instructions. The expression levels of glutamate-cysteine ligase catalytic subunit (GCLC) and glutamate-cysteine ligase modifier subunit (GCLM) mRNA were determined by real time reverse transcription PCR. Measurement data were expressed as Mean ±SD. Comparison between two groups was conducted by independent sample t-test, and comparison among multiple groups was performed by one-way ANOVA. Results:Compared with the radiation group, the proportion of cells with positive staining of SA-β-Gal was significantly lower and cell senescence were alleviated in the treatment 1 and 2 groups (all P<0.001). Compared with the radiation group, the expression level of γH2AX protein was significantly down-regulated ( P=0.037), cell apoptosis rate was significantly decreased ( P=0.026), the proliferation capacity of MLE-12 was enhanced ( P=0.004), GSH ( P=0.002) and SOD ( P<0.001) activity was enhanced and ROS production was declined ( P=0.001) in the treatment 2 group. The expression levels of Nrf2 and p-Nrf2 in total protein were up-regulated over the time of 654-2 intervention. Meanwhile, the expression levels of antioxidant proteins of NQO1 and HO-1 were up-regulated and that of GCLC and GCLM mRNA was also up-regulated. There were no significant differences in the number of cells with positive staining of SA-β-Gal ( P=0.145) and ROS production ( P=0.317) between the inhibitor and radiation groups after supplement of ML385, small-molecule inhibitor of Nrf2. Conclusion:654-2 can activate the Nrf2 pathway, enhance cell antioxidant capacity and inhibit cell senescence, thereby playing a protective role on radiation- induced lung injury.

Objective To prepare specific mouse monoclonal antibody (mAb) against human adenovirus type 55 Hexon protein (HAdV55 Hexon). Methods The Hexon genes of HAdV55, 3, 4, 7, 16 and 21 were chemically synthesized as templates for PCR amplification. The prokaryotic expression plasmids pET28a-HAdV55 Hexon and eukaryotic expression plasmids pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon were constructed respectively. The pET28a-HAdV55 Hexon plasmid was transformed into E. coli competent cell BL21 (DE3) and was induced by IPTG. After the purified inclusion body was denatured and renatured, Hexon55 protein was purified by tangential flow filtration system. pCAGGS-HAdV55 Hexon was used to immunize BALB/c mice by cupping, and HAdV55 Hexon protein was used to booster immunization. The anti-HAdV55 Hexon mAb was prepared by hybridoma technique and the titer and subclass were determined. The specificity of antibody was identified by Western blot using HEK293T cells transfected with pCAGGS-HAdV55 Hexon and by immunofluorescence assay (IFA) using BHK cells transfected with pCAGGS-HAdV55 Hexon. Both clones with high titer were selected, and the cross-reactivity of pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon transfected cells were analyzed by Western blot analysis and IFA. Results PET28a-HAdV55 Hexon and pCAGGS-HAdV55 Hexon, 3, 4, 7, 16 and 21 expression plasmids were successfully constructed. BL21 transformed with pET28a-HAdV55 Hexon was induced by IPTG. The HAdV55 Hexon protein was mainly expressed in the form of inclusion body. After denaturation and renaturation, the purified HAdV55 Hexon protein was obtained by ultrafiltration. Six hybridoma cell lines secreting HAdV55 Hexon mAb were obtained. The antibody subclass analysis showed that 2 strains were IgG2a subtypes and 4 strains were IgG2b. Two specific HAdV55 Hexon antibodies with high titer were obtained, and there was no cross-reactivity with HAdV3, 4, 7, 16, 21 Hexon. Conclusion The specific mice mAb against HAdV55 Hexon provides an experimental basis for establishing its antigen detection method.
Animals ; Mice ; Humans ; Adenoviruses, Human/genetics* ; Escherichia coli/genetics* ; HEK293 Cells ; Isopropyl Thiogalactoside ; Blotting, Western ; Immunoglobulin G ; Antibodies, Monoclonal ; Antibody Specificity ; Mice, Inbred BALB C

Objective:To investigate the protective effect of racanisodamine on lung injury in mice exposed to irradiation.Methods:C57BL/6 mice were randomly divided into control group, racanisodamine group, 18 Gy irradiation group (model group) and racanisodamine combined with 18 Gy irradiation group (treatment group), with 5 mice in each group. The mice in the treatment group received racanisodamine (5 mg/kg) intraperitoneally 3 d before irradiation and contained the whole experiments. Then, single chest irradiation of 18 Gy X-rays was performed both in the model and treatment groups. The racanisodamine group and treatment group received racanisodamine intraperitoneally once a day until 6 weeks after irradiation. The mice were killed at 6 weeks after irradiation. The lung histopathology was observed by HE staining. Serum and bronchial alveolar lavage fluid (BALF) inflammatory cytokines such as TNF-α, IL-1β and IL-6 were determined by ELISA method. Cell senescence was detected by SA-β-Gal staining. The expressions of Nrf2, p-Nrf2 and p62 in lung tissue were performed by immunehistochemistry and Western blot assays.Results:Compared with the model group, the scores of HE staining were decreased ( t=8.66, P<0.01), the number of infiltrated inflammatory cells in BALF were decreased ( t=10.70, P<0.01), and protein concentration in BALF had lower levels ( t=6.75, P<0.01), the serum TNF-α, IL-1β and IL-6 were decreased significantly ( t=8.17, 4.58, 6.54, P<0.01), the activity of SA-β-gal was decreased, and the expressions of Nrf2, p-Nrf2 were enhanced ( t=6.42, 7.30, P<0.01), while the expression of p62 was reduced ( t=4.62, P<0.01) in the treatment group. Conclusions:Racanisodamine plays the protective effect of radiation-induced lung injury by alleviating inflammation associating with the activating of Nrf2-related pathway, which reversed radiation-induced cell senescence.

Objective:Effects of pole-specific acupuncture combined with Bobath on upper limb function, daily life ability and nerve function after traumatic brain injury were observed.Methods:A total of 142 patients with cerebral hemiplegia after traumatic brain injury from January 2019 to December 2020, were divided into the Bobath group (47 cases), the pole-specific acupuncture group (47 cases) and combination group (48 cases) by the random number method. Bobath group received Bobath rehabilitation, the pole-specific acupuncture group received pole-specific acupuncture rehabilitation, and combination group was given pole-specific acupuncture rehabilitation and Bobath treatment. The overall rehabilitation efficiency, limb function Fugl-Meyer scale score, Barthel index of daily life ability, nerve function, and other indicators were observed and compared.Results:After treatment, the overall recovery efficiency (86.96%) in combination group was significantly higher than that of the Bobath group (65.96%) and acupuncture group (64.44%)( χ2=5.84, P=0.016). After treatment, the limb function Fugl-Meyer scale (including upper limb and lower limb function scores)( F=19.38, 24.83, all Ps<0.01), daily life ability Barthel index (including cognitive ability situation score, language ability score, self-care ability score, social adaptability score and total score) of combination group were significantly higher than those in the Bobath group and acupuncture group ( F=14.91, 15.87, 18.71, 18.88, 32.62, all Ps<0.001), while the NIHSS score of combination group was significantly lower than that of the Bobath group and acupuncture group ( F=31.71, P<0.01). After treatment, the NE[(58.29±9.82)μg/L vs. (86.29±12.35)μg/L, (88.34±12.87)μg/L, F=33.39], DA[(204.29±20.26)μg/L vs. (278.72±27.56)μg/L, (281.14±27.82)μg/L, F=55.50], 5-HT[(231.27±20.12)μg/L vs. (294.74±29.34)μg/L, (298.19±28.73)μg/L, F=13.86], E[(21.85±3.19)μg/L vs. (28.37±4.07)μg/L, (28.26±4.14)μg/L, F=9.34] of combination group were significantly lower than those in the Bobath group and acupuncture group ( P<0.01). Conclusion:Magnetic pole-specific acupuncture combined with Bobath can improve the function of limbs, daily quality of life and nerve function of the patients with traumatic craniocerebral injury with cerebral palsy.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Objective:To investigate the molecular genetic mechanisms of cognitive dysfunction in elderly patients with atrial fibrillation.Methods:A case-control study was conducted to analyze statistical data from a previous epidemiological survey, which used a stratified, random cluster sampling method and covered towns and communities in eastern, southern and northern Xinjiang.A total of 8529 residents aged ≥60 years were investigated, of whom, 301 patients with a history of atrial fibrillation or a definitive diagnosis of atrial fibrillation by electrocardiogram during investigation were selected.The patients with atrial fibrillation were divided into a cognitive impairment group(55 cases)and a normal cognitive group(246 cases)according to the diagnostic criteria of cognitive dysfunction.Genotypes of a GWAS on the main atrial fibrillation-related genes were analyzed for genes associated with atrial fibrillation, aging and cognitive impairment.Results:Unconditional multivariate Logistic regression analysis showed that mental work and non-solitary living were protective factors against cognitive impairment in the elderly with atrial fibrillation( OR=0.206, 95% CI: 0.048-0.873; OR=0.286, 95% CI: 0.089-0.922; all P<0.05). Klotho(rs571118)TT and the ApoE ε3/ε4 genotypes were independent risk factors for cognitive impairment in elderly patients with atrial fibrillation( OR=3.922, 95% CI: 1.326-11.595; OR=6.843, 95% CI: 1.263-37.078; all P< 0.05). The interaction between Klotho(rs571118)and ApoE ε3/ε4 was not associated with cognitive impairment in the elderly with atrial fibrillation( OR=1.552, 95% CI: 0.703-3.428; OR=1.897, 95% CI: 0.967-3.723; OR=0.496, 95% CI: 0.061-4.026; all P>0.05). Conclusions:Klotho(rs571118)TT and the ApoE ε3/ε4 genotypes may promote cognitive dysfunction in elderly patients with atrial fibrillation.The results may serve as a basis for research on the mechanisms of cognitive dysfunction in elderly patients with atrial fibrillation.

A total of 120 patients with intermediate coronary artery disease who were admitted to Ankang Hospital of Traditional Chinese Medicine from October 2017 to October 2019 were enrolled in the study (study group), and 120 patients who had ischemic changes on ECG, but no definite angiographic stenosis served as the control group. The SUA and S1P levels were measured, and Gensini scores were evaluated. Coronary blood flow reserve fraction (FFR)≤0.8 was used as the gold standard for diagnosis of functional myocardial ischemia. Patients in the observation group with functional myocardial ischemia and no-functional myocardial ischemia were evaluated. The correlation between variables was analyzed using the Pearson method. The levels of SUA [(492.52±35.47)μmol/L], S1P [(386.42±46.19)nmol/L] and Gensini score (69.42±7.81) in the study group were significantly higher than those in the control group ( t=51.218, 35.227, 60.237; P<0.05).In the study group, patients with functional myocardial ischemia had higher SUA[(678.23±54.21)μmol/L], SIP[(483.45±34.64)nmol/L] and Gensini score[(83.22±5.34) points], and higher incidence of drinking, hypertension, diabetes, and hyperlipidemia compared with no-functional myocardial ischemia patients ( t=24.764, 18.858, 17.888, respectively; χ 2=55.883, 53.684, 27.819, 71.070, 31.111, respectively; P<0.05). Pearson analysis showed that levels of SUA and S1P in patients with coronary artery lesions were positively correlated with Gensini score ( r=0.653, 0.715; P<0.01). The levels of SUA and S1P in patients with intermediate coronary artery lesions are correlated with the severity of coronary stenosis, the combination of the two indicators has better efficacy in predicting functional myocardial ischemia.

Objective: To observe effects of exogenous high mobility group protein box 1 (HMGB1) on angiogenesis in ischemic zone of early scald wounds of rats. Methods: Thirty-six Sprague-Dawley rats were divided into HMGB1 group and simple scald (SS) group according to the random number table, with 18 rats in each group. Comb-like copper mould was placed on the back of rats for 20 s after being immersed in 100 ℃ hot water for 3 to 5 min to make three ischemic zones of wound. Immediately after scald, rats in HMGB1 group were subcutaneously injected with 0.4 μg HMGB1 and 0.1 mL phosphate buffer solution (PBS), and rats in SS group were subcutaneously injected with 0.1 mL PBS from boarders of ischemic zone of scald wound. At post scald hour (PSH) 24, 48, and 72, 6 rats in each group were collected. Protein expressions of vascular endothelial growth factor (VEGF) in ischemic zone of wound at PSH 24, 48, and 72 and protein expressions of CD31 in ischemic zone of wound at PSH 48 and 72 were detected by immunohistochemistry. The number of microvessel in CD31 immunohistochemical sections of ischemic zone of wound at PSH 48 and 72 was calculated after observing by the microscope. The mRNA expressions of VEGF and CD31 in ischemic zone of wound were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction at PSH 24, 48, and 72. Data were processed with analysis of variance of factorial design, t test, and Bonferroni correction. Results: (1) At PSH 24, 48, and 72, protein expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=7.496, 4.437, 5.402, P<0.05 or P<0.01). At PSH 48 and 72, protein expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were 0.038 8±0.007 9 and 0.057 7±0.001 2 respectively, significantly higher than 0.013 4±0.004 9 and 0.030 3±0.004 0 of rats in SS group (t=10.257, 15.055, P<0.01). (2) At PSH 48 and 72, the number of microvessel in ischemic zone of wound of rats in HMGB1 group was obviously more than that of rats in SS group (t=3.536, 4.000, P<0.05). (3) At PSH 24, 48, and 72, mRNA expressions of VEGF in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.406, 3.821, 3.356, P<0.05). At PSH 24 and 48, mRNA expressions of CD31 in ischemic zone of wound of rats in HMGB1 group were significantly higher than those of rats in SS group (t=4.113, 3.466, P<0.05). At PSH 72, mRNA expressions of CD31 in ischemic zone of wound of rats in 2 groups were close (t=0.010, P>0.05). Conclusions Exogenous HMGB1 can promote angiogenesis in ischemic zone of early scald wounds of rats by increasing expressions of VEGF and CD31.

Objective To explore the diagnostic value of tumor markers of CEA,CA153,TPS,Fer and CYFRA21-1 level in postoperative metastasis of breast cancer.Methods After 3-year follow-up for 578 patients with primary breast cancer from 2012 to 2014,we chose 42 patients with metastasis as metastasis group,and another 42 cases without as non-metastasis group.Level of CEA,CA153,TPS,Fer and CYFRA21-1 was detected using chemiluminescence and the difference of CEA,CA153,TPS,Fer and CYFRA21-1 level in 2 groups was compared.Pearson correlation test analysis and ROC curve test were performed for data analysis.Results Serum CEA,CA153,TPS,Fer and CYFRA21-1 levelq6.11 ± 1.87)ng/mL,(26.97 ± 9.52) U/mL,(155.95 ± 22.03)U/L,(173.08 ± 19.87)ng/mL and (18.98 ± 7.33)ng/mL respectively increased in metastasis group when compared with those in non-metastasis group (5.33 ± 1.85)ng/mL,(23.34 ± 8.30)U/mL、(133.22 ± 19.09)U/L,(142.29 ± 22.59) ng/mL and (6.76 ± 1.59)ng/mLrespectively and the change of TPS,Fer and CYFRA21-1 was more obvious and the difference was statistically significant (P ＜ 0.05).There was no significant correlation among serum TPS,Fer and CYFRA21-lin metastasis group.The area under ROC curve of CYFRA21-1 was 0.903,followed by Fer (0.849) and TPS (0.791) and in the diagnosis of metastasis of breast cancer,CYFRA21-1 has great diagnostic value.Conclusions The increase of serum TPS,Fer and CYFRA21-1 level is closely associated with postoperative metastasis of breast cancer and CYFRA21-1 has great diagnostic value for monitoring postoperative metastasis of breast cancer.