ZEB2 knockdown inhibits interleukin-1ß-induced cartilage degradation and inflammatory response through the Wnt/ß-catenin pathway in human chondrocytes.

OBJECTIVE: Osteoarthritis (OA) is a degenerative disease of the joints characterized by inflammation and cartilage degeneration. Zinc finger E-box binding homeobox 2 (ZEB2) contains various function domains that interact with multiple transcription factors involved in various cellular functions. However, the function of ZEB2 in OA has not been clearly illustrated. METHOD: Interleukin-1ß (IL-1ß) was used to establish an OA model in vitro. We quantified the ZEB2 expression in cartilage tissues from OA patients and IL-1ß-induced chondrocytes through reverse transcription-quantitative polymerase chain reaction and Western blot. We then used functional assays to explore the function of ZEB2 during OA progression. RESULTS: ZEB2 expression was increased in OA cartilage tissues and chondrocytes. The silencing of ZEB2 increased aggrecan and collagen II levels, and reduced the content of matrix metalloproteinase-3 (MMP-3), MMP-9, and MMP-13. ZEB2 knockdown inhibited the effects of IL-1ß on the production of nitric oxide and prostaglandin E2, and the expression of inducible nitric oxide synthase and cyclooxygenase-2. ZEB2 inhibition also suppressed the levels of IL-6 and tumour necrosis factor-α, and increased the IL-10 level in IL-1ß-treated cells. Mechanically, ZEB2 knockdown blocked the activation of the Wnt/ß-catenin pathway in chondrocytes. CONCLUSION: Knockdown of ZEB2 alleviated IL-1ß-induced cartilage degradation and the inflammatory response through the Wnt/ß-catenin pathway in chondrocytes.

[Current challenges in the diagnosis and treatment of filamentous mycosis in critically ill patients].

Influenza and coronavirus disease 2019 are independent risk factors for the development of invasive fungal disease (IFD) in critically ill patients in the intensive care unit. IFD, particularly mold infections, have a high mortality rate. The diagnosis and treatment of mold infections is challenging, and early detection and timely treatment are crucial in reducing the mortality of IFD. This review will summarize the latest epidemiology and risk factors for the development of mold infections in critically ill patients, current diagnostic criteria and challenges, as well as the treatment strategies recommended by the international clinical guidelines, aiming to provide some references for the diagnosis and treatment of mold infections in critically ill patients in clinical practice.

[Applications and challenges of large language models in critical care medicine].

The rapid development of big data methods and technologies has provided more and more new ideas and methods for clinical diagnosis and treatment. The emergence of large language models (LLM) has made it possible for human-computer interactive dialogues and applications in complex medical scenarios. Critical care medicine is a process of continuous dynamic targeted treatment. The huge data generated in this process needs to be integrated and optimized through models for clinical application, interaction in teaching simulation, and assistance in scientific research. Using the LLM represented by generative pre-trained transformer ChatGPT can initially realize the application in the diagnosis of severe diseases, the prediction of death risk and the management of medical records. At the same time, the time and space limitations, illusions and ethical and moral issues of ChatGPT emerged as the times require. In the future, it is undeniable that it may play a huge role in the diagnosis and treatment of critical care medicine, but the current application should be combined with more clinical knowledge reserves of critical care medicine to carefully judge its conclusions.

[Establishment of a prognostic model for non-nephrotic membranous nephropathy based on unbalanced data].

Objective: To explore the construction of a machine learning model based on unbalanced data to predict the progression of non-nephrotic membranous nephropathy. Methods: The clinical and pathological data of patients diagnosed with non-nephrotic membranous nephropathy by renal biopsy in Shanxi People's Hospital from January 2018 to December 2021 were retrospectively analyzed.The prediction models were constructed based on logistic regression, support vector machine (SVM) and light gradient boosting machine (lightGBM), respectively. The mixed sampling technology was used to process the unbalanced data, and the area under the receiver operating characteristic curve (AUC) was used to evaluate the predictive performance of the models. Finally, Shapley additive explanation (SHAP) was used to interpret the results of the optimal prediction model. Results: A total of 148 patients were included in the study, including 84 males and 64 females, with a mean age of (47.2±12.5) years. The follow-up time [M(Q1, Q3)] was 14(7, 20) months. Twenty-three patients (15.5%) achieved the renal end-point event in the study. The SVM model had the highest AUC (0.868, 95%CI: 0.813-0.925), followed by logistic regression (AUC=0.865, 95%CI: 0.755-0.899) and lightGBM (AUC=0.791, 95%CI: 0.690-0.882). The feature recursive elimination cross validation (RFECV) method based on random forest (RF) and the SHAP plot based on the SVM model showed that immunohistochemistry IgG, total protein (TP), anti-phospholipase A2 receptor (anti-PLA2R), blood chloride and D-Dimer were risk factors affecting the progress of non-nephrotic membranous nephropathy. Moreover, patients with high immunohistochemistry IgG, anti-PLA2R and D-Dimer had an increased risk of achieving the renal end-point event. Conclusion: The SVM model established in this study can effectively predict the progress of non-nephrotic membranous nephropathy, and provide a new method for the early identification of high-risk patients and precision therapy.

[Analysis of enterovirus infection type among acute respiratory tract infection cases in Luohe City, Henan Province from 2017 to 2021].

Objective: To understand the infection status of Enterovirus (EV) in cases of acute respiratory infections (ARIs) in Luohe City, Henan Province from 2017 to 2021, and analyze the prevalence and type composition of EV in ARIs. Methods: From October 2017 to May 2021, pharyngeal swab samples were collected from 1 828 patients with ARIs in Luohe Central Hospital and the clinical epidemiological data of these cases were also collected. EV-positive samples were identified by Quantitative Real-time Polymerase Chain Reaction (qPCR). The 5'-untranslated region (5'UTR) was amplified by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). The results of 5'UTR region were initially typed by Enterovirus Genotyping Tool Version 1.0. Based on the typing results, the full-length of VP1 region was amplified by RT-PCR. The EV typing was identified again by VP1 region. Results: Among 1 828 cases of ARIs, 56.7% (1 036) were males. The median (Q1, Q3) age was about 3 (1, 5) years. Patients under 5 years old accounted for 71.6% (1 309 cases). Among all cases, a total of 71 EV-positive samples were identified by qPCR, with a detection rate of 3.88% (71/1 828). The EV detection rates for men and women were 3.28% (34/1 036) and 4.67% (37/792), without statistically significant differences (χ2=2.32, P=0.14). The EV detection rates for 2 to <6 years, 6 months to <2 years, 6 to <10 years, and <6 months were 6.29% (48/763), 3.00% (18/600), 2.52% (4/159), and 1.67% (1/60) (χ2=27.91, P<0.001). The EV detection rate was 0.92% (3/326) in autumn and winter of 2017. The EV detection rates were 1.18% (6/508), 2.47% (12/485) and 8.31% (34/409) in each year from 2018 to 2020, with an increasing trend year by year(χ2trend=29.76, P<0.001). The main prevalent seasons were summer and autumn. The detection rate in spring of 2021 was 4.00% (4/100). A total of 12 types were identified and classified as CVA2, CVA4, CVA5, CVA6, CVA10, CVB3, CVB5, E5, E11, E30, PV-1, and EV-D68. The types of CVA2, CVA10, CVA6, and CVB3 were the dominant phenotypes. In 59 sample of EV typing, the main clinical manifestation was upper respiratory tract infection (36/59, 61.01%). The dominant types detected in upper respiratory tract infections were CVA10 (10/36, 27.78%), CVA6 (9/36, 25.00%) and CVB3 (8/36, 22.22%). The dominant type detected in lower respiratory tract infections was CVA2 (7/19, 36.84%). Conclusion: In Luohe City, Henan Province from 2017 to 2021, EV infection in ARIs cases has clear seasonal and age-specific patterns, and the dominant types of upper and lower respiratory tract infections are different.

[Study on the comparative analysis of the efficacy of transmesenteric vein extrahepatic portosystemic shunt and transjugular intrahepatic portosystemic shunt in the treatment of cavernous transformation of portal vein].

Objective: To compare the safety and efficacy of transmesenteric vein extrahepatic portosystemic shunt (TEPS) and transjugular intrahepatic portosystemic shunt (TIPS) in the treatment of cavernous transformation of the portal vein (CTPV). Methods: The clinical data of CTPV patients with patency or partial patency of the superior mesenteric vein treated with TIPS or TEPS treatment in the Department of Vascular Surgery of Henan Provincial People's Hospital from January 2019 to December 2021 were selected. The differences in baseline data, surgical success rate, complication rate, incidence rate of hepatic encephalopathy, and other related indicators between TIPS and TEPS group were statistically analyzed by independent sample t-test, Mann-Whitney U test, and Chi-square test. Kaplan-Meier survival curve was used to calculate the cumulative patency rate of the shunt and the recurrence rate of postoperative portal hypertension symptoms in both groups. Results: The surgical success rate (100% vs. 65.52%), surgical complication rate (6.67% vs. 36.84%), cumulative shunt patency rate (100% vs. 70.70%), and cumulative symptom recurrence rate (0% vs. 25.71%) of the TEPS group and TIPS group were statistically significantly different (P < 0.05). The time of establishing the shunt [28 (2141) min vs. 82 (51206) min], the number of stents used [1 (12) vs. 2 (15)], and the length of the shunt [10 (912) cm vs. 16 (1220) cm] were statistically significant between the two groups (t = -3.764, -4.059, -1.765, P < 0.05). The incidence of postoperative hepatic encephalopathy in the TEPS group and TIPS group was 6.67% and 15.79% respectively, with no statistically significant difference (Fisher's exact probability method, P = 0.613). The pressure of superior mesenteric vein decreased from (29.33 ± 1.99) mmHg to (14.60 ± 2.80) mmHg in the TEPS group and from (29.68 ± 2.31) mmHg to (15.79 ± 3.01) mmHg in TIPS group after surgery, and the difference was statistically significant (t = 16.625, 15.959, P < 0.01). Conclusion: The best indication of TEPS is in CTPV patients with patency or partial patency of the superior mesenteric vein. TEPS improves the accuracy and success rate of surgery and reduces the incidence of complications.

[Clinical characteristics of 272 437 patients with different histopathological subtypes of primary esophageal malignant tumors].

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 χ2 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.

[Reality and expectation of machine learning in critically ill patients with acute kidney injury].

The prediction model constructed by machine learning can early predict acute kidney injury (AKI) in critically ill patients, which contributes to taking preventive measures to reduce incidence of AKI as soon as possible. Machine learning can also identify AKI subtypes in real time, so as to facilitate the implementation of precise treatments for their subtypes in the future, which may improve the prognosis of patients. In this article, the construction of forecasting model of AKI and identification of AKI subtypes in critically ill patients and it's precise treatment with machine learning, limitations and development trend of machine learning in the field of AKI in critically ill patients is addressed for clinical reference.

[Ventilatory management in shared airway surgery: current status, challenges, and future directions].

A shared airway during anesthesia refers to maintaining the airway and ventilation of the patient by the anesthesiologist as the surgeon performs the procedures in the same anatomic space. Due to the narrow airway space in shared airway surgery, the difficulty lies both in the exposure of surgery field and implementation of the optimized airway management strategy and maintaining the airway safety in clinical practice. In recent years, many new visualized airway management tools and supraglottic ventilation devices have been invented and verified in clinical settings. Especially, the development of tubeless supraglottic oxygenation and ventilation technique provides new insights into the airway management. This review intends to provide a reference for future innovations in airway management tools and ventilation techniques by summarizing previous airway management strategies of shared airway surgery to benefit more patients.

[Analysis of the short-and medium-term curative effect of TIPS approach combined with AngioJet thrombus aspiration technology treatment in acute portal vein thrombosis].

Objective: To evaluate the short- and medium-term clinical efficacy of TIPS approach combined with AngioJet thrombus aspiration technology treatment in acute portal vein thrombosis. Methods: 63 cases with acute portal vein thrombosis treated in our center from May 2017 to July 2019 were studied retrospectively, including 49 males and 14 females, aged 35-61 (46 ± 5) years. TIPS approach (with/without) combined with Angiojet thrombus aspiration and gastroesophageal varices embolization was performed simultaneously according to the patient's condition. Regular follow-up for 3-33 (22 ± 3) months after surgery was used to observe the curative effect. Results: The technical success rate was 100%. Portal vein and superior mesenteric vein blood flow were returned to normal after the operation. Two cases of biliary tract injury were untreated. Simultaneously, two cases of intrahepatic arteriovenous fistula were treated with superselective arterial embolization. During the follow-up period, 47 cases (74.61%) had complete portal vein recanalization, 13 cases (20.63%) had partial recanalization, 3 cases (4.76%) had complete portal cavernoma, 7 cases (11.11%) had symptomatic hepatic encephalopathy, 1 case had received artificial liver treatment (1.59%), 1 case had peptic ulcer (11.11%), 6 cases (9.52%) had lost to follow-up, and there was no portal hypertension-related bleeding or death. Conclusion: TIPS approach combined with AngioJet thrombus aspiration technology is safe, effective and feasible in the treatment of acute portal vein thrombosis, and the short- and medium-term clinical effects are satisfactory.

Identification of a novel splice variant isoform of interferon regulatory factor 10, IRF10, in orange spotted grouper Epinephelus coioides.

Interferon regulatory factors (IRFs) are a family of transcription factors essential to the control of antiviral immune response, cell growth, differentiation and apoptosis. IRF10 was originally found in chicken, which was induced by the v-Rel oncoprotein in lymphoid cell lines and involved in the upregulation of major histocompatibility complex (MHC) class I and guanylate-binding protein. In fish, IRF10 plays negative roles in regulation of the interferon (IFN) response. Here, we identified a splice variant of IRF10, named as EcIRF10-SF in orange spotted grouper, which shares the first three exons with the long form (EcIRF10-LF) and retains part of intron 3, creating a premature termination codon. Furthermore, we observed that the EcIRF10-SF exhibits similar expression pattern compared to its native counterparts. Functional studies demonstrate that the two EcIRF10 isoforms repress DrIFNϕ1 and DrIFNϕ3 promoter activity and negatively regulate fish antiviral gene expression. Subcellular localization analysis shows that the amino acids from 57 to 86 within DBD are required for IRF10 nuclear import. Overall, our description of transcript diversification of IRF10 in the grouper provides a coherent framework to further dissect its roles in immune response.

Upregulation of SNHG12 accelerates cell proliferation, migration, invasion and restrain cell apoptosis in breast cancer by enhancing regulating SALL4 expression via sponging miR-15a-5p.

Breast cancer (BC) is malignant cancer that threatens the health of millions of females worldwide. Long non-coding RNA (lncRNA) small nucleolar RNA host gene 12 (SNHG12) has been identified as an oncogene in multiple cancers. However, the regulatory role of SNHG12 in BC cell progression is still obscured. The levels of SNHG12, miR-15a-5p, and Sal-like 4 (SALL4) in BC tumor tissues and cells were measured by qRT-PCR. Cell viability, apoptosis, migration, and invasion were examined by CCK8, flow cytometry, and transwell assay, respectively. The interaction between miR-15a-5p and SNHG12 or SALL4 was evaluated by dual-luciferase reporter assay. Protein expression of SALL4 was analyzed by western blot. Xenograft mice were established by subcutaneously injecting BC cells stably transfected with sh-SNHG12 and sh-NC. SNHG12 and SALL4 expressions were upregulated whereas miR-15a-5p was downregulated in BC tumors compared with normal tissues. Besides, miR-15a-5p was correlated with SNHG12 and SALL4 inversely as calculated by Pearson's correlation coefficient. More importantly, SNHG12 knockdown attenuated BC tumor growth in vitro and in vivo. Subsequently, dual-luciferase reporter assay confirmed the interaction between miR-15a-5p and SNHG12 or SALL4. The rescue experiments revealed that miR-15a-5p inhibitor restored SNHG12 silencing induced inhibition on BC cell proliferation, migration, invasion, and promotion of apoptosis. Additionally, SNHG12 was found to accelerate BC cell progression by absorbing miR-15a-5p to enhance SALL4 expression. SNHG12 promotes cell proliferation, migration, and invasion but suppresses apoptosis in BC by upregulating SALL4 expression via sponging miR-15a-5p, representing potential targets for the development of novel diagnosis and treatment methods.

[Investigation on demands for antenatal care services among 2 002 pregnant women during the epidemic of COVID-19 in Shanghai].

Objective: To identify problems and demands for antenatal care (ANC) among pregnant women in different trimesters of pregnancy in Shanghai for optimizing ANC service during the epidemic of COVID-19. Method: s Organized by maternal and child health care institute in the 16 districts of Shanghai, a cross sectional study was conducted among pregnant women who came to pregnancy registration in the community health centers or attended ANC in midwifery hospitals from February 7 to February 12, 2020. Consented participating women completed a semi-structured online questionnaire voluntarily. Data was analyzed using frequency,chi-square test and scoring. Result: s A total of 2 002 valid questionnaires were collected from 183 community health centers and 67 midwifery hospitals. About 94.6%(1 894/2 002) of the pregnant women worried about being infected during the COVID-19 epidemic, and 14.7% (294/2 002) demanded for psychological consultation. Time-lapse appointments for ANC were requested by 87.7% (1 756/2 002) of the participants for avoiding presenting themselves in people-density places. Compared with other pregnancy trimesters, pregnant women in the second trimester were more willing to reduce the frequency of ANC (35.2% versus 39.5% versus 48.1%, P<0.01). Compared with multiparas, primiparas were more willing to have online consultation and guidance (49.2% versus 63.8%, P<0.01). Regarding the needs for health knowledge on COVID-19, personal protection against 2019 novel coronavirus (2019-nCoV) was the most concerned for pregnant women, and 71.0% (1 421/2 002) of them preferred to obtain knowledge through health applications, official Weibo and WeChat. Conclusions: Pregnant women in Shanghai critically concern about the risk of 2019-nCoV infections, and highly demand knowledge and measures on prevention and protection from COVID-19. They ask for having time-lapse appointments for ANC and online access to health information and services. Maternal and child care institutes should understand the demands of pregnant women, optimize the means of ANC service, and provide tailored and accessible health education and service for the safety of mother and child.

[The role of three-dimensional speckle tracking imaging derived parameters on predicting outcome of hypertrophic cardiomyopathy patients with MYH7 mutations].

Objective: To evaluate the cardiac functional changes in hypertrophic cardiomyopathy(HCM) patients with ß-myosin heavy chain gene (MYH7) mutations by three-dimensional (3D) speckle tracking imaging(3D-STI) and conventional echocardiography modalities, and then to explore the potential predictors of adverse cardiovascular events in these patients. Methods: A consecutive series of 192 HCM patients admitted in our center from October 2014 to October 2016 were genetically screened to identify MYH7 mutations in this retrospective study. A total of 43 HCM patients with MYH7 mutations were enrolled. The patients were divided into events group(n=13) and no event group(n=30) according to the presence or absence of adverse cardiovascular events(primary and secondary endpoints). All patients were followed up to January 2019 after comprehensive evaluation of 3D-STI, two-dimensional and Doppler echocardiography. The adverse cardiovascular events were recorded. Results: The median follow up time was 1 012 (812, 1 330) days. During follow-up, 13 patients (30.2%) reached endpoints: 6 cases of the primary endpoints(2 cases of sudden cardiac death(SCD), 3 cases of survival after defibrillation, and 1 case of appropriate implantable cardioverter-defibrillator(ICD) discharge); 7 cases of the second endpoints(5 cases of heart failure hospitalization, 1 case of syncope and cardioversion due to supraventricular tachycardia, and 1 case of end-stage HCM). Patients with adverse cardiovascular events had higher prevalence of syncope and risk of SCD, enlarged left atrial volume index(LAVI) and reduced 3D left ventricular global longitudinal train (3D-GLS), as compared to those without adverse events(all P<0.05). The multivariate Cox regression analysis showed that reduced 3D-GLS(HR=0.814, 95%CI 0.663-0.999, P=0.049) was an independent predictor for adverse cardiovascular events. The cutoff value of 3D-GLS≤13.67% was linked with significantly increased risk of adverse cardiovascular events in this patient cohort(AUC=0.753, 95%CI 0.558-0.948, sensitivity 86%, specificity 69%, P<0.05). The Kaplan-Meier analysis indicated that the patients with the 3D-GLS≤ 13.67% faced higher risk of death than those with 3D-GLS>13.67%. Conclusion: 3D-GLS is useful on predicting adverse cardiovascular events in HCM patients with MYH7 mutations.

Scaffolding adapter protein Gab1 impairs bFGF-induced bio-activity via affecting PI3K-AKT pathway.

The role of Grb2-associated binder 1 (Gab1) in bFGF-activated PI3K-AKT pathway of endothelial cells remains largely unknown. To elucidate this role, a set of studies with siRNA knockdown of Gab1 was performed. Knockdown of Gab1 using siRNA was performed in fused endothelial cell line EA.hy926 and the low level of Gab1 was confirmed with quantitative R-T PCR and Western blotting. Effects of Gab1 down-regulation were examined on several aspects: bFGF-induced AKT phosphorylation, proliferation, migration and vessel tubing formation of EA.hy926 cells. The bFGF-induced AKT phosphorylation of wild-type EA.hy926 cells was both dose-dependent and time dependent with a peak at 10 ng/ml and about 30 min after bFGF treatment. The AKT activation was significantly reduced in Gab1 siRNA-treated EA.hy926 cells. The blocking of Gab1-AKT path resulted in a set of biological alterations of EA.hy926 cells: (i) reduced proliferation; (ii) impaired migration; (iii) decreased vessel tubing formation in both 2D and 3D culture. All data support that Gab1 is associated with angiogenesis function of EA.hy926 endothelium cells via PI3K-Akt signaling pathway.

Characterization and phylogenomics of the complete mitochondrial genome of the polyzoic cestode Gangesia oligonchis (Platyhelminthes: Onchoproteocephalidea).

The order Onchoproteocephalidea (Eucestoda) was recently erected to accommodate the hook-bearing tetraphyllideans and the proteocephalideans, which are characterized by internal proglottization and a tetra-acetabulate scolex. The recognized subfamilies in the Proteocephalidae appeared to be non-monophyletic based on 28S recombinant DNA (rDNA) sequence data. Other molecular markers with higher phylogenetic resolution, such as large mitochondrial DNA fragments and multiple genes, are obviously needed. Thus the mitochondrial genome of Gangesia oligonchis, belonging to the putative earliest diverging group of the Proteocephalidae, was sequenced. The circular mitogenome of G. oligonchis was 13,958 bp in size, and contained the standard 36 genes: 22 transfer RNA genes, two rRNA genes and 12 protein-coding genes, as well as two major non-coding regions. A short NCR and a large NCR (lNCR) region were 216 bp and 419 bp in size, respectively. Highly repetitive regions in the lNCR region were detected with that of 11 repeat units. The mitogenome of G. oligonchis shared 71.1% nucleotide identity with Testudotaenia sp. WL-2016. Phylogenetic analyses of the complete mitochondrial genomes with Bayesian inference and maximum likelihood methods indicated that G. oligonchis formed a sister clade with Testudotaenia sp. WL-2016 with maximum support. The ordinal topology is (Caryophyllidea, (Diphyllobothriidea, (Bothriocephalidea, (Onchoproteocephalidea, Cyclophyllidea)))). The mitogenomic gene arrangement of G. oligonchis was identical to that of Testudotaenia sp. WL-2016. Both mitogenomic and nuclear sequence data for many more taxa are required to effectively explore the inter-relationships among the Onchoproteocephalidea.

[Effects of thoracic epidural administration of lidocaine on hemodynamic and arousal responses of double lumen tracheal intubation during induction of anesthesia].

OBJECTIVE: To compare the effects of thoracic epidural administration of lidocaine on hemodynamic and arousal responses of double lumen tracheal intubation during induction of anesthesia. METHODS: In the study, 40 patients with American Society of Anesthesiologists (ASA) physical statuses I-II, aged 19-66 years, scheduled for elective thoracic surgeries under general anesthesia requiring orotracheal intubation were allocated to either the double-lumen endobronchial intubation (T group) or double-lumen endobronchial intubation after epidural administration of lidocaine (E group). After an intravenous anesthetic induction, the orotracheal double-lumen intubation was performed using a Macintosh direct laryngoscopy (MDLS), respectively. Invasive blood pressure (BP), heart rate (HR) and bispectral index (BIS) were recorded before and after anesthetic induction, immediately after intubation and 5 minutes after intubation with 1-minute interval and the intubation time also noted. The rate pressure product (RPP) was calculated. RESULTS: After anesthetic induction, BP and RPP in the two groups decreased significantly compared with their preinduction values. In comparison with their postinduction values, the orotracheal intubation in the two groups caused significant increases in BPs, HRs and RPP. In comparison with their preinduction values, BPs decreased significantly in E group, systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) increased significantly and lasted for 1 min in T group. The HRs of both groups after intubation were significantly higher than their baseline values , and increased in HR and lasted for 1 min and 4 min in E group and T group, respectively. SBP, DBP, MAP, HR and RPP after intubation in T group were significantly higher than those of E group during the observation period. The values of BIS were similar between both the groups. In T group, the incidences of SBP percent increased>30% of the baseline value and RPP more than 22 000 were significantly higher than in E group. None of the patients in group E had SBP more than 130% of the baseline value and RPP more than 22 000. CONCLUSION: During double-lumen endobronchial intubation, epidural administration of lidocaine can provide less hemodynamic response and similar arousal response.

Genetic polymorphism of human leucocyte antigen and susceptibility to multidrug-resistant and rifampicin-resistant tuberculosis in Han Chinese from Hubei Province.

We determined the high-resolution allele and haplotype frequencies at the human leucocyte antigen (HLA)A, B and DRB1 loci in the Han population of Hubei province, the TB endemic area of Central China, with pulmonary tuberculosis (PTB), and established the relationship between HLA-A, B and DRB1 alleles as well as haplotypes and susceptibility to multidrug-resistant and rifampicin-resistant tuberculosis (MDR/RR-TB). Blood samples were drawn from 174 patients with MDR/RR-TB and 838 patients with drug-susceptible PTB in ethnic Han population from Hubei province (central China). Four-digit allele genotyping of HLA- A, B and DRB1 loci was performed using polymerase chain reaction with sequence-specific oligonucleotide probes (PCR- SSOP). The allele and haplotype frequencies of HLA-A, B and DRB1 were determined and compared between patients with MDR/RR-TB and patients with drug-susceptible PTB. Statistical analysis of the generated data indicated no departure from expectation of Hardy-Weinberg equilibrium (HWE) at all loci of the control group. Multivariate analysis identified allele DRB1*08:01 (p < .0001; OR = 174.5, 95% CI 15.3-1987.2) as independent predictor of MDR/RR-TB, except for old age (p < .0001; OR = 10. 9, 95% CI 7.6-15.8), previous treatment history (p < .0001; OR = 11.0, 95% CI 7.2-16.7) and poor compliance to treatment (p < .0001; OR = 12.9, 95% CI 8.4-20.0). While in the subgroup of new TB cases, DRB1*08:01 (p < .0001; OR = 80.3, 95% CI 7.0-917.1) and older age (p < .0001; OR = 3.9, 95% CI 2.4-6.4) were independent susceptibility factors for primary MDR/RR-TB. Our results suggest that a combination of clinical and host genetic information about tuberculosis patients may contribute to prediction and early detection of MDR/RR-TB.

Sequencing, characterization and phylogenomics of the complete mitochondrial genome of Dactylogyrus lamellatus (Monogenea: Dactylogyridae).

Despite the worldwide distribution and pathogenicity of monogenean parasites belonging to the largest helminth genus, Dactylogyrus, there are no complete Dactylogyrinae (subfamily) mitogenomes published to date. In order to fill this knowledge gap, we have sequenced and characterized the complete mitogenome of Dactylogyrus lamellatus, a common parasite on the gills of grass carp (Ctenopharyngodon idella). The circular mitogenome is 15,187 bp in size, containing the standard 22 tRNA genes, 2 rRNA genes, 12 protein-encoding genes and a long non-coding region (NCR). There are two highly repetitive regions in the NCR. We have used concatenated nucleotide sequences of all 36 genes to perform the phylogenetic analysis using Bayesian inference and maximum likelihood approaches. As expected, the two dactylogyrids, D. lamellatus (Dactylogyrinae) and Tetrancistrum nebulosi (Ancyrocephalinae), were closely related to each other. These two formed a sister group with Capsalidae, and this cluster finally formed a further sister group with Gyrodactylidae. Phylogenetic affinity between Dactylogyrinae and Ancyrocephalinae was further confirmed by the similarity in their gene arrangement. The sequencing of the first Dactylogyrinae, along with a more suitable selection of outgroups, has enabled us to infer a much better phylogenetic resolution than recent mitogenomic studies. However, as many lineages of the class Monogenea remain underrepresented or not represented at all, a much larger number of mitogenome sequences will have to be available in order to infer the evolutionary relationships among the monogeneans fully, and with certainty.

[The effect of DNA hydroxymethylase Tet2 on γ globin activation in the treatment of ß-thalassemia].

Objective: To study the function of ten-eleven translocation 2 (Tet2) in γ globin gene expression in patients with ß- thalassemia. Methods: Gamma globin expression was induced by 5-azacytidine and Tet2 gene expression was knocked down by short hairpin RNA (shRNA) in a human immortalized myelogenous leukemia K562 cell line. The global 5-hydroxymethylcytosine (5hmC) level was measured by an ELISA kit. 5hmC level of γ globin gene was quantified by sulfite sequencing. The mRNA level of Tet2, γ globin, and related transcription factors Nfe4 and Klf1 were quantified by real-time PCR. Results: Tet2 knockdown resulted in a decreased global 5hmC level from 0.14% to 0.03% as of the control group in K562 cells. The expression of γ globin was enhanced after 5-azacytidine treatment in vitro. However, γ globin mRNA level in Tet2 knockdown cells was only 55% as that in control group. The CG sites on γ globin gene were unmethylated. As Tet2 was down-regulated, the expression levels of Nfe4 and Klf1 decreased by about 80% and increased to 3.5 folds, respectively. Conclusions: Tet2 appears to maintain 5hmC level and facilitates γ globin gene activation. Moreover, Tet2 more likely regulates γ globin expression via affecting transcription factors rather than the gene itself. Thus, Tet2 could be a potential therapeutic target for ß thalassemias.