BACKGROUND:Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy.Therefore,exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.OBJECTIVE:To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques,in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.METHODS:Two datasets of pulmonary fibrosis,GSE24206 and GSE110147,were downloaded from the Gene Expression Omnibus(GEO)database(a public database developed and maintained by the U.S.National Center for Biotechnology Information to store and share bioinformatics data),and the gene expression matrices of these two datasets were normalized by using the"limma"package in R software.The autophagy-related genes were extracted from GeneCards database(a database created by the U.S.National Center for Biotechnology Information,which automatically integrates gene-centric data from about 200 Web sources,including genomic,transcriptomic,proteomic,genetic,clinical,and functional information).Differential gene analysis was performed on the pulmonary fibrosis dataset,and the common genes were extracted by cross-comparing the differential genes with the autophagy genes,so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis.The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes.Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning,and core genes were subjected to the enrichment analysis.Diagnostic models were constructed from the identified core genes.Calibration curves were used to assess the predictive ability of the line graph model.An external dataset,GSE21369,was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy,as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database.Finally,human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment,and the relative expression of genes in the model cells was verified using qRT-PCR.RESULTS AND CONCLUSION:(1)A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained.Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue,collagen metabolism,collagen pro-fibroblasts,and growth factor binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions.(2)Immunoinfiltration analysis revealed that the expression of activated memory CD4+T cells,M0 macrophages,and resting dendritic cells was significantly elevated in the pulmonary fibrosis group(P＜0.05),showing a strong correlation.(3)Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified:COL1A2 and IL6.The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately,and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance.COL1A2 and IL6 were related to the cell-cycle pathway,mitogen-activated protein kinase signaling pathway,Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions.A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes,and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi.COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis.To conclude,COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis,but its specificity to pulmonary fibrosis needs to be further investigated.

BACKGROUND:Early diagnosis of pulmonary fibrosis is the foundation for timely antifibrotic drug therapy.Therefore,exploring and discovering ideal biomarkers that can be effectively used for the early diagnosis of pulmonary fibrosis is crucial for the treatment of the disease.OBJECTIVE:To conduct an in-depth analysis of key autophagy-related genes involved in the process of pulmonary fibrosis by means of bioinformatics and machine learning techniques,in order to investigate whether autophagy-related core genes of pulmonary fibrosis can be used as reliable biomarkers in the assessment of the progression of pulmonary fibrosis.METHODS:Two datasets of pulmonary fibrosis,GSE24206 and GSE110147,were downloaded from the Gene Expression Omnibus(GEO)database(a public database developed and maintained by the U.S.National Center for Biotechnology Information to store and share bioinformatics data),and the gene expression matrices of these two datasets were normalized by using the"limma"package in R software.The autophagy-related genes were extracted from GeneCards database(a database created by the U.S.National Center for Biotechnology Information,which automatically integrates gene-centric data from about 200 Web sources,including genomic,transcriptomic,proteomic,genetic,clinical,and functional information).Differential gene analysis was performed on the pulmonary fibrosis dataset,and the common genes were extracted by cross-comparing the differential genes with the autophagy genes,so as to identify autophagy genes that may play a role in the process of pulmonary fibrosis.The intersecting genes were analyzed for functional enrichment and cellular immune infiltration by gene ontology and Kyoto Encyclopedia of Genes and Genomes.Core genes of pulmonary fibrosis associated with autophagy were screened by protein-protein interactions and machine learning,and core genes were subjected to the enrichment analysis.Diagnostic models were constructed from the identified core genes.Calibration curves were used to assess the predictive ability of the line graph model.An external dataset,GSE21369,was used to perform a receiver operating characteristic curve analysis to validate the expression profiles of pulmonary fibrosis genes associated with autophagy,as well as to predict Chinese herbs associated with the genes IL6 and COL1A2 via the Coremine database.Finally,human embryonic lung fibroblasts were cultured and modelled by transforming growth factor-β1 treatment,and the relative expression of genes in the model cells was verified using qRT-PCR.RESULTS AND CONCLUSION:(1)A total of 51 pulmonary fibrosis differential genes and 25 genes intersecting with autophagy genes were obtained.Gene ontology analysis showed that the 25 intersecting genes were related to extracellular matrix tissue,collagen metabolism,collagen pro-fibroblasts,and growth factor binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes enrichment analysis indicated that they were mainly related to the Phosphatidylinositol 3-kinase/protein kinase B signaling pathway and the signaling pathway of the extracellular matrix-receptor interactions.(2)Immunoinfiltration analysis revealed that the expression of activated memory CD4+T cells,M0 macrophages,and resting dendritic cells was significantly elevated in the pulmonary fibrosis group(P＜0.05),showing a strong correlation.(3)Two autophagy signature genes involved in the progression of pulmonary fibrosis were identified:COL1A2 and IL6.The column-line diagram model showed that the two core genes predicted the onset of pulmonary fibrosis more accurately,and the receiver operating characteristic curve analysis showed that the two characteristic genes had diagnostic significance.COL1A2 and IL6 were related to the cell-cycle pathway,mitogen-activated protein kinase signaling pathway,Janus kinase-signal transduction and activator of transcription signaling pathway and cytokine-cytokine receptor interactions.A total of 20 Chinese herbs were predicted to be related to COL1A2 and IL6 genes,and their efficacies were mainly to clear away heat and detoxify toxins and to invigorate blood and move qi.COL1A2 and IL6 were verified to be highly expressed in pulmonary fibrosis.To conclude,COL1A2 and IL6 may be potential diagnostic biomarkers for pulmonary fibrosis,but its specificity to pulmonary fibrosis needs to be further investigated.

Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).

Objective : To investigate the expression characteristics of mu opioid receptor ( MOR) in rat colon smooth muscle cells by cultured rat primary colonic smooth muscle cells . Methods : colonic smooth muscle cells were isolated , cultured and identified ; immunofluorescence double labeling method was used to observe the distribution characteristics of MOR , Endomorphin⁃2 (EM2) , and calmodulin (CaM) in colonic smooth muscle cells ; Western Blot method was used to detect the expression of MOR and EM2 in smooth muscle cells of the colon . After the intervention measures Acetylcholine (ACh) ( 1 × 10 - 3 mol/L) and EM2 (2 μmol/L) were applied , the changes of CaM protein expression were observed ; The calciumion imaging method was used to detect the changes of calciumi on concentration in smooth muscle cells. Results: The colonic smooth muscle cells were cultured and identified . The positive cells labeled with α ⁃smooth muscle actin ( α ⁃SMA) accounted for more than 95% of the total number of cells . Immunofluorescence double labeling showed that there were MOR and EM2 distributions in colonic smooth muscle cells , and all MOR and EM2 positive cells coexisted with α ⁃SMA . Western Blot results showed that there were MOR and EM2 expressions in colonic smooth muscle cells . CaM in the ACh group significantly increased at 10 minutes (P < 0. 05) , CaM in the EM2 group significantly decreased ( P < 0. 05) ; The calciumion imaging results showed that alone applied ACh , the calciumion concentration in smooth muscle cells significantly increased ( P <0. 05) ; Alone applied EM2 , the calciumion concentration in colonic smooth muscle cells was down⁃regulated (P <0. 05) ; Applied ACh and EM2 sequentially , EM2 significantly reduced the increase of the calciumion concentration in smooth muscle cells induced by ACh (P < 0. 05) . Conclusion MOR and EM2 are expressed in colonic smooth muscle cells , and EM2 may inhibit the expression of CaM and reduce the concentration of calcium ions through MOR .

OBJECTIVE To study the protective effect of phenolic acid components from Salvia deserta Schang on the oxidative stress injury of human renal tubular epithelial cells HK -2 induced by high glucose and high fat . METHODS HK-2 cells were divided into control group ，model group ，canagliflozin group （positive control group ，15 μmol/L），purified product of phenolic acids from S. deserta Schang group （10.8 μg/mL），4 monomers group （salvianic acid ，protocatechuic aldehyde ，caffeic acid，rosmarinic acid ，50 μmol/L）. In addition to the control group ，cell injury model of high glucose and high fat was established in other groups （500 μmol/L palmitic acid+ 30 mmol/L glucose for 48 h）and cultured for 48 h. The cell apoptotic rate ，the contents of malondialdehyde （MDA）and glutathione （GSH），and the activity of superoxide dismutase （SOD）were detected in each group ； the expression levels of nuclear erythroid 2-related factor 2（Nrf2），Kelch-like ECH -associated protein 1（Keap1）protein，heme oxygenase-1（HO-1）and NADH ：quinone acceptor oxidoreductase 1（NQO1）were determined in above 5 groups（except for salvianic acid ，protocatechuic aldehyde ，caffeic acid ）. RESULTS Compared with control group ，the apoptotic rate of HK -2 cells in model group was increased significantly （P＜0.01）；the content of MDA was increased significantly （P＜0.01），while the content of GSH and the activity of SOD were decreased significantly ；protein expressions of Nrf 2，NQO1 and HO -1 2018D01C169） were significantly down -regulated（P＜0.01），while the protein expression of Keap 1 was up -regulated significantly （P＜0.01）. Compared with model group ， the apoptotic rate and the content of MDA were decreased significantly in administration groups（P＜0.01）；the content of GSH in administration groups and the activity of SOD in purified product of phenolic acids group，protocatechuic aldehyde group and rosmarinic acid group were increased significantly （P＜0.01）. The protein expressions of Nrf2，HO-1 and NQO 1 in purified product of phenolic acids group as well as the protein expression of Nrf 2 in rosmarinic acid group were up -regulated significantly （P＜0.01），while the protein expression of Keap 1 was down -regulated significantly in purified product of phenolic acids group and rosmarinic acid group （P＜0.01）. CONCLUSIONS The phenolic acids components from S. deserta Schang can relieve oxidative stress injury of renal tubular epithelial cells induced by high glucose and high fat ，the mechanism of which may be associated with activating Keap 1/Nrf2 signaling pathway and inhibiting oxidative stress response .

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

Objective:To investigate the predictive value of 18F-fluorodeoxyglucose (FDG) PET/CT imaging for the epidermal growth factor receptor (EGFR) mutations in patients with lung adenocarcinoma. Methods:From January 2013 to December 2017, a total of 146 patients (83 males, 63 females, age: (60.2±10.3) years) who were confirmed as lung adenocarcinoma by pathology and were examined by 18F-FDG PET/CT imaging and EGFR mutation testing in Shanxi Cancer Hospital were retrospectively analyzed. The differences of clinical characteristics (age, gender, smoking, tumor diameter, loymph node metastasis, distant metastasis, stage, thyroid transcripition factor-1 (TTF-1), NapsinA, cyiokeratin (CK)-7, Ki-67) and PET/CT parameters (maximun standardized uptake value (SUV max) of the primary tumor (pSUV max), SUV max of lymph node (nSUV max) and SUV max of distant metastasis (mSUV max)) between patients of EGFR mutation and EGFR wild type were analyzed using independent-sample t test, χ2 test and Fisher exact test. The predictors for EGFR mutation were analyzed by logistic regression analysis. The predictive value of pSUV max and pSUV max combined with gender, smoking and tumor diameter was determined by receiver operating characteristic (ROC) curve analysis. Results:There were 46.58%(68/146) patients with EGFR mutations and 53.42%(78/146) patients with wild type. Gender, smoking, lymph node metastasis, tumor diameter, pSUV max, nSUV max, TTF-1, NapsinA and Ki-67 were significantly different between patients with EGFR mutations and those with wild type ( t values: from -3.023 to -2.032, χ2 values: 4.725-33.749, all P<0.05). Female (odds ratio ( OR)=3.236, 95% CI: 1.213-8.779; P=0.029), non-smoker ( OR=4.947, 95% CI: 1.796-13.621; P=0.019), tumor diameter<3.5 cm ( OR=2.750, 95% CI: 1.109-6.818; P=0.001) and pSUV max<9.1( OR=2.960, 95% CI: 1.227-7.141; P=0.016) were predictors of EGFR mutations in lung adenocarcinoma. The area under the curve (AUC) of pSUV max was 0.640 with the specificity of 43.6%(34/78)and the sensitivity of 27.9%(19/68), while the AUC of the four independent factors was 0.83 with the specificity of 71.8%(56/78) and the sensitivity of 83.8%(19/68). Conclusions:pSUV max is associated with mutant EGFR status. Moreover, the combination of pSUV max, gender, smoking and tumor diameter can enhance the predictive value on EGFR mutation status in patients with lung adenocarcinoma.

Objective To investigate the efficacy of pranoprofen drops on dry eye of patients with Sj(o)gren's syndrome (SS).Methods This is a prospective study.Sixty-eight inpatients with dry eye in our hospital were randomly divided into the experimental and control groups.Right eyes were taken for the trial,with 34 cases in each group.The experimental group was given pranoprofen eye drops combined with polyethylene glycol eye drops.Eyes of the control group were given polyethylene glycol drops only.Corneal fluorescein staining (FL),tear film breakup time (BUT) and Schirmer test (SIT) were tested before treatment and 1,2,4 weeks after treatment by the same care giver.The levels of IL-6 and TNF-α in tears were detected by ELISA.Analysis of variance of repeated data and t test were used for statistical analysis.Results The difference of FL,BUT,SIT and content IL-6 and TNF-α in tears in the experimental group patients before treatment and 1,2,4 weeks after treatment were signifcant (F=4.65,7.53,6.43,9.96,10.87; P＜0.05),which were statistically significantly different between the experimental group and the control group patients (F=3.27,5.85,4.36,8.36,7.23; P＜0.05).One week after treatment and before treatment,the difference of BUT and SIT of the two groups was not statistically significant (P＞0.05),those of the 2 weeks after treatment were statistically significantly different [BUT of the experimental group was (11.1±2.5) s,BUT of the control group was (9.7±1.9) s,t=2.594 8,P＜0.05; the SIT of the experimental group was (7.3±1.7) mm,the SIT of the control group was (5.9±1.7) mm,t=3.571 8,P＜0.05].BUT of the two groups at 4 weeks after treatment was statistically significantly different [BUT of the experimental group was (14.4±2.8) s,BUT of the control group was (11.4±2.6) s,t=4.469 4,P＜0.05; the SIT of the experimental group was (9.9±2.1) mm,the SIT of the control group was (8.7±1.9) mm,t=2.568 0,P＜0.05].The difference of FL and IL-6 and TNF-α in tears pretreatment between the two groups was not statistically significant (P＞0.05).At week 1,2,4 after treatment,the differences between the two groups were statistically significant (tFL=4.173 9,3.190 7,4.072 6; tIL-6=2.131 5,2.316 4,5.310 1; tTNF-α=2.216 4,4.871 9,8.175 0; P＜0.05).No significant discomfort and side effects were observed in the two groups.Conclusion Pranoprofen drops can significantly improve symptoms of dry eye in patients with pSS,in particular,the repair of the cornea,may be related to the inhibition of the expression of ocular inflammatory cytokines IL-6 and TNF-α,and thus reduce the ocular surface inflammatory reaction.

Objective To isolate and identify the putative Japanese encephalitis virus (JEV) re-ceptors from C6/36 and Vero cells. Methods Molecules binding with JEV were isolated from C6/36 and Vero cells by co-immunoprecipitation (Co-IP) approach, identified by mass spectrometry, and detected by Western blot. The location of putative JEV receptor on cells membrane and the binding with JEV were ob-served by laser scanning confocal microscopy (LCM). Results Several molecules binding with JEV were isolated from C6/36 and Vero cells by Co-IP, and only one molecule was identified as heat shock cognate 70 (HSC70) by mass spectrometry. Antibody against HSC'70 was able to detect a 74 ×103 protein isolated by Co-IP from C6/36 and Vero cells membrane in Western blot assays. It was observed by LSCM that when JEV attached on the surface of C6/36 cells, JEV and HSCT0 protein were co-localization. Conclusion 74 x 103 molecular identified as HSC70 protein from C6/36 cells may be JEV receptor.

OBJECTIVETo analyze the value of the diagnostic criteria for bladder outlet obstruction in benign prostatic hyperplasia (BPH).

METHODSA total of 358 patients with BPH were divided into 3 grades according to fibrous urethrocystoscopy information on the severity of obstructions, which were classified as Grade 1 (slight), Grade 2 (moderate), and Grade 3 (severe). By Schäfer's graph they were divided into 7 grades, represented by 0 to VI. We analyzed the volume of prostate, maximum flow rate (Qmax), residual urine volume, International Prostatic Symptom Score (IPSS) and detrusor instability. Statistical analysis ANOVA (analysis of variance) was made, spearman correlation evaluated and the coefficient of determination measured.

RESULTSOf all the patients, 27 were classified as Grade 1, 236 as Grade 2 and 95 as Grade 3. Eighty-four patients had detrusor instability. The volumes of the prostate ranged from 16 ml to 145 ml, averaging (47.04 +/- 15.61) ml. The mean maximum flow rate was (10.02 +/- 2.12) ml/min and the mean residual urine volume was (84.06 +/- 36.50) ml. With the increase of the severity of obstruction, the volume of the prostate increased (F = 4.216, P < 0.05), IPSS rose (F = 8.408, P < 0.001), the maximum flow rate decreased (F = 22.43, P < 0.001), the residual urine volume rose (F = 163.232, P < 0.001), the incidence of detrusor instability increased (F = 23.637, P < 0.001) and Schäfer's grades were elevated (F = 202.897, P < 0.001). The volume of the prostate, the maximum flow rate (Qmax), residual urine volume, IPSS detrusor instability and Schäfer's grades were all correlated significantly with the severity of the obstruction. The correlation index and coefficient of determination were r = 0.29, R2 = 0.08; r = 0.35, R2 = 0.12; r = -0.69, R2 = 0.47; r = 0.60, R2 = 0.36; r = 0.33, R2 = 0.11; r = 0.72, R2 = 0.52; respectively. The correlation between the urethrocystoscopy information and Schäfer's graph on the severity of the obstruction were the best criteria of all.

CONCLUSIONThe severity of the obstruction at urethrocystoscopy correlates well with that at urodynamic investigation. Such criteria could improve the sensitivity and specificity of the diagnosis of bladder outlet obstruction.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; complications ; diagnosis ; Retrospective Studies ; Urinary Bladder Neck Obstruction ; diagnosis ; etiology ; Urodynamics