Myocardial strain analysis by 4D-speckle tracking echocardiography for prediction of coronary artery disease severity in patients with stable angina pectoris

Objective: We sought to evaluate the myocardial strain by four-dimensional speckle-tracking echocardiography (4D-STE) in patients with stable angina pectoris (SAP) to determine the severity of coronary artery disease (CAD) based on the Gensini score. Methods: The present study comprised of 150 patients with SAP. Patients with history of SAP, normal left ventricular ejection fraction, and without regional wall motion abnormalities (RWMA) were scheduled for elective coronary angiography. Based on Gensini score, there were two groups: non-critical stenosis group [Gensini score (0e19), n ¼ 117] and critical stenosis group [Gensini score 20, n ¼ 33]. Correlation between Gensini score and 4D-STE strain parameters were investigated. Results: Out of 150 patients, critical stenosis group had significantly depressed values of all 4D-STE strain parameters than non-critical stenosis group (p < 0.001), except global radial strain (GRS) parameter. Significant positive correlation was found between Gensini score and 4D global longitudinal strain (GLS), global circumferential strain (GCS), global area strain (GAS) with Spearman's correlation coefficient (r) as 0.626, 0.548, and 0.631, respectively (p < 0.001), whereas significant negative correlation was found between Gensini score and GRS (r ¼ 0.433, p < 0.001). A 4D GLS value of 17 had 84.9% sensitivity and 97.4% specificity, GAS 31 (90.9% sensitivity, 78.6% specificity), GCS 17 (69.7% sensitivity, 92.3% specificity), and GRS <47 (sensitivity 72.7%, specificity 76.1%) to detect critical CAD described by Gensini score 20. Conclusion: The 4D-STE can aid in the assessment of severe CAD stenosis with good sensitivity and specificity in the patients with SAP without RWMA on traditional echocardiography.

Construction of a stable centromere protein F overexpression cell model of hepatocellular carcinoma using CRISPR activation system / 生物工程学报

Current studies have shown that centromere protein F (CENPF) was overexpressed in hepatocellular carcinoma (HCC) and might be involved in the pathogenesis of HCC. Specifically, due to the very large molecular weight (358 kDa) of CENPF full length protein, only CENPF knock-down, but not overexpression models, were applied currently to explore the carcinogenicity of CENPF in HCC. Whether CENPF overexpression is a cause or an effect in HCC remains to be illustrated. We aimed to establish a CENPF overexpression cell model using CRISPR/dCas9 synergistic activation mediator (SAM) system with lentiMPHv2 and lentiSAMv2 vectors to explore the role of CENPF overexpression in HCC. Single guide RNAs (sgRNAs) that specifically identify the transcription initiation site of CENPF gene were synthesized and inserted into the lentiSAMv2 plasmid. Huh-7 and HCCLM3 cells were first transduced with lentiMPHv2 and then selected with hygromycin B. The cells were then transduced with lentiSAMv2 carrying specific sgRNA for CENPF gene, followed by blasticidin S selection. The mRNA and protein detection results of Huh-7 and HCCLM3 cells screened by hygromycin B and blasticidin S showed that the endogenous overexpression of CENPF can be induced by sgRNA1 and sgRNA4, especially by sgRNA4. By using the CRISPR/dCas9 technique, stable cell models with overexpressed CENPF were successfully constructed to explore the role of CENPF in tumorigenesis, which provides a reference for the construction of cell models overexpressing large molecular weight protein.

Efficacy and Safety of Fruquintinib Combined with Sintilimab in Treatment of Advanced Microsatellite Stable Colorectal Cancer / 肿瘤防治研究

Objective To investigate the clinical efficacy and safety of fruquintinib combined with sintilimab in the treatment of advanced microsatellite stable (MSS) colorectal cancer. Methods A retrospective study of 44 patients with MSS colorectal cancer treated with fruquintinib and sintilimab was conducted.The patients were divided into the fruquintinib alone (n=22) and fruquintinib combined with sintilimab (n=22) groups.The treatment regimen was as follows: The patients in the fruquintinib alone group consumed oral fruquintinib capsules at 5 mg/d once for three consecutive weeks with a one week stop in 28 day cycles.The patients in the fruquintinib combined sintilimab group were injected intravenously with sintilimab (200 mg) once per three weeks, and fruquintinib was used in the same manner as the fruquintinib alone group. Results The objective response rate (ORR) of the fruquintinib alone group was 9.09%, the disease control rate (DCR) of the fruquintinib alone group was 45.45%.The ORR of the fruquintinib combined with sintilimab group was 18.18%, and the DCR was 63.64%.The median PFS of the fruquintinib alone and fruquintinib combined with sintilimab groups were 4.4 months (IQR: 2.1-8.2) and 6.7 months (IQR: 3.9-12.6), respectively (χ2=4.372, P=0.037).Most of the adverse reactions during the treatment of the two groups were grades 1-2.In addition, no significant difference in the incidence of adverse reactions was found between two groups (P > 0.05). Conclusion Compared with fruquintinib alone, fruquintinib combined with sintilimab in the treatment of patients with MSS colorectal cancer after the failure of standard treatment has better clinical efficacy, and adverse drug reactions can be controlled.

Network Meta-analysis of Chinese medicine injection combined with conventional western medicine in treatment of stable angina pectoris / 中国中药杂志

This study aimed to systematically evaluate the efficacy and safety of different Chinese medicine injections combined with conventional western medicine for stable angina pectoris. PubMed, Cochrane Library, EMbase, Web of Science, CNKI, Wanfang, VIP, and SinoMed were searched to collect randomized controlled trial(RCT) of Chinese medicine injection combined with conventio-nal western medicine in the treatment of stable angina pectoris from the inception of the databases to July 8, 2022. Two researchers independently screened the literature, extracted the data, and evaluated the risk of bias of the included studies. Stata 15.1 was used for network Meta-analysis. A total of 52 RCTs were included, involving 4 828 patients treated by 9 Chinese medicine injections(Danhong Injection, Salvia Miltiorrhiza Polyphenol Hydrochloride Injection, Tanshinone Sodium Ⅱ_A Sulfonate Injection, Salvia Miltiorrhiza Ligustrazine Injection, Dazhu Hongjingtian Injection, Puerarin Injection, Safflower Yellow Pigment Injection, Shenmai Injection and Xuesaitong Injection). The network Meta-analysis showed that:(1)in terms of improving the efficacy of angina pectoris, the surface under the cumulative ranking curve(SUCRA) followed the order of conventional western medicine combined with Salvia Miltiorrhiza Ligustrazine Injection>Tanshinone Sodium Ⅱ_A Sulfonate Injection>Danhong Injection>Salvia Miltiorrhiza Polyphenol Hydrochloride Injection>Xuesaitong Injection>Shenmai Injection>Puerarin Injection>Safflower Yellow Pigment Injection>Dazhu Hongjingtian Injection;(2)in terms of improving the efficacy of electrocardiogram(ECG), SUCRA followed the order of conventional western medicine combined with Salvia Miltiorrhiza Ligustrazine Injection>Puerarin Injection>Danhong Injection>Salvia Miltiorrhiza Polyphenol Hydrochloride Injection>Shenmai Injection>Xuesaitong Injection>Safflower Yellow Pigment Injection>Tanshinone Sodium Ⅱ_A Sulfonate Injection>Dazhu Hongjingtian Injection;(3)in terms of increasing high-density lipoprotein cholesterol(HDL-C), SUCRA followed the order of conventional western medicine combined with Danhong Injection>Shenmai Injection>Safflower Yellow Pigment Injection>Xuesaitong Injection>Tanshinone Sodium Ⅱ_A Sulfonate Injection>Dazhu Hongjingtian Injection;(4)in terms of lowering low-density lipoprotein cholesterol(LDL-C), SUCRA followed the order of conventional western medicine combined with Safflower Yellow Pigment Injection>Danhong Injection>Shenmai Injection>Tanshinone Sodium Ⅱ_A Sulfonate Injection>Dazhu Hongjingtian Injection>Xuesaitong Injection;(5)in terms of safety, the overall adverse reactions of Chinese medicine injection combined with conventional western medicine were less than those of the control group. Current evidence indicated that Chinese medicine injection combined with conventional western medicine could improve the curative effect of stable angina pectoris with higher safety. Limited by the number and quality of included studies, the above conclusion needed to be verified by more high-quality studies.

Discrimination of cultivation modes of Dendrobium nobile based on content of mineral elements and ratios of nitrogen stable isotopes / 中国中药杂志

This study explored the feasibility of mineral element content and ratios of nitrogen isotopes to discriminate the cultivation mode of Dendrobium nobile in order to provide theoretical support for the discrimination of the cultivation mode of D. nobile. The content of 11 mineral elements(N, K, Ca, P, Mg, Na, Fe, Cu, Zn, Mn, and B) and nitrogen isotope ratios in D. nobile and its substrate samples in three cultivation methods(greenhouse cultivation, tree-attached cultivation, and stone-attached cultivation) were determined. According to the analysis of variance, principal component analysis, and stepwise discriminant analysis, the samples of different cultivation types were classified. The results showed that the nitrogen isotope ratios and the content of elements except for Zn were significantly different among different cultivation types of D. nobile(P<0.05). The results of correlation analysis showed that the nitrogen isotope ratios, mineral element content, and effective component content in D. nobile were correlated with the nitrogen isotope ratio and mineral element content in the corresponding substrate samples to varying degrees. Principal component analysis can preliminarily classify the samples of D. nobile, but some samples overlapped. Through stepwise discriminant analysis, six indicators, including δ~(15)N, K, Cu, P, Na, and Ca, were screened out, which could be used to establish the discriminant model of D. nobile cultivation methods, and the overall correct discrimination rates after back-substitution test, cross-check, and external validation were all 100%. Therefore, nitrogen isotope ratios and mineral element fingerprints combined with multivariate statistical analysis could effectively discriminate the cultivation types of D. nobile. The results of this study provide a new method for the identification of the cultivation type and production area of D. nobile and an experimental basis for the quality evaluation and quality control of D. nobile.

Progress of Biomarkers to Predict Response to Immune Checkpoint Inhibitors in Microsatellite Stability Colorectal Cancer / 肿瘤防治研究

The exploration of biomarkers predicting response to immune checkpoint inhibitors in microsatellite stability colorectal cancer can enable more patients to benefit from immunotherapy. Tumor mutational burden (TMB), POLE/POLD1 mutation, CMS classifications, MGMT methylation, and other indicators own the potential and value of predicting response to immune checkpoint inhibitors in microsatellite stability colorectal cancer. In this paper, we reviewed the related research on predictive biomarkers of immune checkpoint inhibitors in microsatellite stability colorectal cancer, provide a reference for the best treatment strategy for microsatellite stability colorectal cancer.

Research progress on tumor microenvironment and immune combination therapy of MSS colorectal cancer / 国际肿瘤学杂志

In recent years, immunotherapy, especially immune checkpoint inhibitors, has shown obvious advantages in prolonging the survival of patients with advanced tumors, and the tumor microenvironment is one of the important factors affecting the efficacy of immunity. Patients with microsatellite-stable colorectal cancer exhibit immune responses in combination with immune checkpoint inhibitor therapy. In-depth exploration of the tumor microenvironment characteristics of microsatellite-stable colorectal cancer and the application of combined immune checkpoint inhibitor therapy can provide new ideas and directions for colorectal cancer immunotherapy.

Finite Element Analysis on Different Internal Fixations for Treating Proximal Humeral Fractures in Children / 医用生物力学

Objective To compare biomechanical characteristics of external fixator, Kirschner’s wire, elastic stable intramedullary nailing (ESIN) for fixing proximal humeral fractures in children by finite element method.Methods The CT scanning data from the healthy humerus of an 8-year-old patient with proximal humeralfractures were collected, and the image data were imported in Mimics 21. 0 to establish the rough humeralmodel, which was imported in Geomagic 2013 to construct the three-dimensional (3D) model of cancellous and cortical bones of the humerus. After the model was assembled with 3 fixators ( external fixator, Kirschner’swire, ESIN), it was imported in ANSYS 2019 to simulate the upper limb under quiet, abduction, adduction, flexion, extension, external rotation, internal rotation working conditions. The maximum displacement of the distal humerus, the maximum stress of the fixture, and the maximum displacement of the distal fracture surface were analyzed. Results The minimum values of the maximum displacement of the distal humerus in models fixed by external fixator, Kirschner’s wire, ESIN appeared under extension (2. 406 mm), external rotation (0. 203 mm), external rotation (0. 185 mm) working conditions, respectively. Conclusions External fixator is the most unstable fixation of proximal humeral fractures in children, and the biomechanical performance of ESIN is better than that of external fixator and Kirschner’s wire fixation

Acupuncture and moxibustion in treatment of chronic obstructive pulmonary disease at stable stage: a network Meta-analysis / 中国针灸

The efficacy on chronic obstructive pulmonary disease (COPD) at stable stage treated with different methods of acupuncture and moxibustion was evaluated using network Meta-analysis method. The articles of the randomized controlled trial (RCT) on stable COPD treated with acupuncture and moxibustion were searched electronically in CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase, Web of Science and Cochrane library. The search was conducted from the inception of the databases to March 20th, 2022. Data analysis was performed using R4.1.1, Stata16.0 and RevMan5.3 softwares. A total of 48 RCTs were included, involving 15 kinds of acupuncture and moxibustion interventions and a sample size of 3 900 cases. The results of network Meta-analysis showed that: ① For the forced expiratory volume in one second predicted (FEV1%), both the governor vessel moxibustion combined with conventional treatment (G+C therapy) and the yang-supplementing moxibustion combined with conventional treatment (Y+C therapy) obtained the better effect than that of the conventional treatment (P<0.05), and the G+C therapy was more effective compared with the thread-embedding therapy combined with conventional treatment (E+C therapy) and warm needling (P<0.05). ② Concerning to COPD assessment test (CAT) score, the results indicated that the Y+C therapy, and the mild moxibustion combined with conventional treatment (M+C therapy) were more effective when compared with the conventional treatment (P<0.05), and the effect of the Y+C therapy was better than that of the E+C therapy (P<0.05). ③ Regarding six-minute walking distance (6MWD), the effect of acupuncture combined with conventional treatment (A+C therapy) was better than that of either the E+C therapy or the conventional treatment (P<0.05). The effect of the G+C therapy was optimal for improving FEV1%, the Y+C therapy obtained the best effect for improving CAT score, and A+C therapy was the most effective for improving 6MWD. Due to the limitation of the quality and quantity of included studies, this conclusion needs to be further verified through high-quality RCT.

Stable Angina Pectoris with Sleep Disorders Treated by Modified Tongmai Anshen Formula （通脉安神方加减）： An Clinical Observation of 74 Cases / 中医杂志

ObjectiveTo observe the clinical efficacy of the Modified Tongmai Anshen Formula （通脉安神方加减， MTAF） in the treatment of stable angina pectoris （SAP） with sleep disorders. MethodsA total of 148 patients suffering from SAP with sleep disorder were included and randomly divided into control group and treatment group， with 74 patients in each group. The control group received conventional western medicine， and the treatment group additionally received MTAF （1 dose per day）， both for 4 weeks. The changes in angina pectoris symptoms， traditional Chinese medicine （TCM） syndromes， sleep quality， quality of life， serological indicators including serum intercellular adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule-1 （VCAM-1）， brain-derived nerve growth factor （BDNF） and tyrosine kinase receptor B （TrkB） were compared between groups before and after treatment， and the safety was evaluated. ResultsIn the treatment group and the control group， the total effective rates of TCM syndromes（82.43% vs 52.70%）， angina pectoris （79.73% vs 64.86%） and sleep （89.19% vs 68.92%） showing significant difference （P＜0.001）. After treatment， the total TCM syndrome score， primary symptom score， secondary symptom score， and secondary symptoms sleeplessness， restlessness， tiredness and fatigue individual score， angina pectoris score， PSQI total score and each item score were all significantly reduced in both groups， while the SF-36 single item score significantly increased （P＜0.05）. The total TCM syndromes and primary symptom scores， secon-dary symptoms sleeplessness， restlessness， tiredness and fatigue individual score， angina pectoris score， time to fall asleep， sleep quality， hypnotic medication， sleep disturbance， daytime dysfunction score and PSQI total score were significantly lower in the treatment group than those in the control group after treatment （P＜0.05）， while the somatic pain， general health status， social functioning， emotional functioning， mental health， and health change were significantly higher in the treatment group （P＜0.05）. After treatment， ICAM-1 and VCAM-1 level significantly decreased （P＜0.05）， and BDNF and TrkB levels increased （P＜0.05） in the treatment group， while BDNF level significantly decreased in the control group （P＜0.05）. The TrkB level was significantly higher in the treatment group compared to the control group after treatment （P＜0.05）. A total of four adverse events occurred during the treatment， none of which were considered to be related to this study. ConclusionMTAF can significantly improve angina pectoris symptoms， TCM syndromes， sleep quality and quality of life in patients suffering from SAP with sleep disorders， the mechanism of which may be related to the protection of vascular endothelial function and central neurons.

Clinical outcomes of selecting the vertebra above sagittal stable vertebra as distal fusion level for Scheuermann's thoracic kyphosis / 中华骨科杂志

Objective:To investigate the clinical outcome and complications associated with utilizing sagittal plane stable vertebra-1 (SSV-1) as the distal instrumented vertebra (LIV) in posterior fusion of thoracic kyphosis with Scheuermann's Disease kyphosis (STK).Methods:A longitudinal study on patients with STK who underwent posterior correction and fusion surgery from January 2018 to June 2021 were conducted. All participants had a follow-up duration over two years. Patients were divided into two groups according to the segment of LIV: the SSV group, where LIV was located in SSV; and the SSV-1 group, where LIV was located in the vertebral body above SSV. The radiographic parameters, including global kyphosis (GK), lumbar lordosis (LL), and sagittal plane (SVA), LIV offset distance (LIV translation), pelvic incidence (PI), pelvic tilt (PT) and sacral slope (SS), were compared between the two groups. The SRS-22 scale was used to evaluate health-related quality of life at pre-operation and last follow-up, and the incidence of postoperative distal junctional kyphosis (DJK) was also recorded. Analytical techniques, such as Analysis of Variance and Mann-Whitney tests, were employed to compare inter-group differences.Results:A total of 57 patients were included in the study, 36 in the SSV group and 21 in the SSV-1 group. The average age for patients were 16.1±2.3 years (range 13-20 years), and the average follow-up time was 32.8±6.8 months (range 24-53 months). There were no statistically significant differences between the two groups in terms of gender, age, follow-up time, surgical time, intraoperative bleeding volume, and fusion level. Before surgery, the LIV deviation distance in the SSV group was significantly lower than that in the SSV-1 group (-7.9±11.0 mm vs. 31.5±11.5 mm, t=7.64, P<0.001). In the SSV group, the preoperative GK was 79.3°±10.5°, and the last follow-up GK was 44.4°±8.5°, which was significantly improved compared to preoperative value ( t=28.28, P<0.001); in the SSV-1 group, the preoperative GK was 81.1°±10.6°, and the value at 1-week post-operative was 44.9°±7.8°, which was significantly improved compared to pre-operative value ( t=22.23, P<0.001). At the last follow-up, it was 45.1°±8.7°, with a correction rate of 44.3%±8.5%. No significant difference was observed between the two groups in terms of GK, LL, SVA, PI, PT and SS at pre-operative, 1-week post-operative and last follow-up ( P>0.05). All patients had no intraoperative complications of nerve injury. During the follow-up period, one patient (1/21, 4.8%) developed DJK without complications such as proximal kyphosis, pseudarthrosis, or failed internal fixation. At the last follow-up, the functional score of SRS-22 in SSV-1 group improved from preoperative (3.5±0.54) to postoperative (4.1±0.62), with an average improvement rate of 19.2%±3.2%, and the difference was statistically significant ( t=3.74, P=0.001). These results indicating that the surgical treatment was effective in relieving the symptoms of the patients. Conclusion:Selecting SSV-1 as LIV in corrective surgeries for STK appears to produce commendable clinical results with minimal implant-associated complications over a two-year observation period.

Research progress on stable gene transfection methods of Plasmodium falciparum / 中国热带医学

@#Abstract: Transfection of Plasmodium falciparum is helpful to study the function of its genes, such as drug resistance. However, transgenic manipulation has been very challenging, mainly due to the high A/T base sequence structure (A+T content of about 82%) and low transfection efficiency of the Plasmodium genome. Electroporation-based transfection of Plasmodium falciparum has been successfully applied in the study of certain genes, and electroporation by preloading is currently the preferred method for introducing foreign DNA into Plasmodium falciparum. The site-directed editing of Plasmodium genes mostly adopts the method of two-plasmid transfection. It is generally believed that successful transfection of Plasmodium requires a large amount of high-purity plasmid DNA and an accurate transfection system. In addition to the evaluation of the current commonly used electrotransfection methods, this paper also introduces a new transfection method, namely lyse-reseal erythrocytes for transfection (LyRET). This paper also review the role of factors such as plasmid DNA concentration, the use of transfection reagents, the setting of transfection parameters, the addition of fresh red blood cells, and the markers of successful transfection in improving the success rate and efficiency of Plasmodium transfection, in the hope of providing a reference for study in this field.

Taking the mechanism of glucose catabolism disorder in depression as an example to explore the research ideas and strategies of stable isotope tracer metabolomics / 药学学报

Stable isotope tracer metabolomics tracks and analyzes the whole metabolic process of the body through the tracer atoms, which belongs to the frontier technology in the field of biomedicine. This technology is of great significance and value for explaining the pathogenesis of diseases, finding biomarkers of diseases and drug action targets. Taking the mechanism of glucose catabolism disorder in depression as an example, this paper systematically expounds the stable isotope tracer metabolomics technology and its application. The research idea of stable isotope tracer metabolomics based on unmarked metabolomics was put forward, and the research strategy of biological significance interpretation from four dimensions of metabolite isotope abundance, key metabolic enzymes, metabolic flow direction and metabolite flow was given, which broke through the bottleneck of stable isotope tracer metabolomics research technology based on overall animal experiment, and provided scientific basis for the promotion and application of this technology.

Clinical observation of chemotherapy combined with camrelizumab in patients with inoperable microsatellite stable type colorectal cancer / 中国药房

OBJECTIVE To evaluate the clinical efficacy and safety of XELOX chemotherapy （oxaliplatin+capecitabine） combined with antiangiogenic agent （apatinib） and immunotherapy （camrelizumab） in patients with inoperable metastatic colorectal cancer （CRC）of microsatellite stable （MSS） type. METHODS Clinical medical records of 40 patients with inoperable metastatic CRC of MSS type treated in Lishui People’s Hospital from January 2020 to January 2021 were retrospectively collected. According to the treatment plan， the patients were divided into control group （20 cases） and observation group （20 cases）. Control group was given XELOX+apatinib regimen， while observation group was given XELOX+apatinib+camrelizumab regimen. Every 3 weeks was a treatment cycle， and the treatment lasted for 2 consecutive cycles. The objective response rate （ORR）， disease control rate （DCR）， progression-free survival （PFS）， overall survival （OS） and adverse events （AEs） were recorded for all patients. RESULTS The ORR and DCR of observation group were 65.0% and 85.0%， respectively； and the ORR and DCR of control group were 35.0% and 75.0%， respectively， with no statistical significance between 2 groups （P＞0.05）. The median PFS of observation group and control groups were 16.0 months and 8.0 months， respectively； and the median OS were 19.0 months and 12.5 months， respectively， with statistical significance between 2 groups （P＜0.05）. Each patient in both groups had at least one AEs， and the incidences of reactive skin capillary hyperplasia and hyperthyroidism in observation group （40.0%， 20.0%） were significantly higher than those in control group （both were 0） （P＜0.05）. The incidence of nausea and vomiting in control group （90%） was significantly higher than observation group （10%） （P＜0.05）. There were 14 cases （70.0%） of patients with grade 3 or above AEs in observation group， and only 5 cases （25.0%） in control group， with statistical significance between 2 groups （P＜0.05）. However， no severe AEs that could not be tolerated or fatal occurred in the two groups， which could be alleviated after drug withdrawal or treatment. CONCLUSIONS The efficacy of XELOX chemotherapy combined with apatinib and camrelizumab in inoperable metastatic CRC patients of MSS type is comparable to that of XELOX chemotherapy combined with apatinib， but it has certain advantages in ORR， PFS and OS， and controllable safety.

Construction of the human hepatocellular carcinoma cell line stably expressed mutated hepatitis B virus X genes and changes of its biological behavior / 中国热带医学

@#Abstract: Objective　To construct HepG2, Huh7 cell lines stably express hepatitis B virus X (HBx) mutant (C1653T, T1753C), and explore their effect on the biological behavior of hepatocellular carcinoma cells. Methods　The lentivirus plasmid of pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were obtained by PCR site mutagenesis according to wild type ayr HBx. Double enzyme digestion and Sanger sequencing were performed for accuracy of plasmid. Blank HepG2 and Huh7 cells were used as the control group, HepG2, Huh7 cells were infected by pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, and pLVX-HBxT1753C-IRES-tdTomato lentivirus solution, then monoclonal cell was selected by 0.6 μg/mL puromycin. Immunostaining and Western Blot were performed for the verification of stable strains. CCK8 assay was performed for the proliferation capacity of stable strains. Western Blot was performed for expression of EMT-related signal molecules in cells. The independent samples t-test was used for comparison between two groups. Results Double enzyme digestion and Sanger sequencing showed that that the size of the cut fragments of recombinant lentiviral plasmids was correct, and the point mutation location and base substitution were correct, suggesting that the plasmid of pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were constructed successfully. Immunostaining and Western blot showed that HBX were expressed in stable strains, while there was no HBX expression in the blank control group, indicating that the HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed. CCK8 assay showed that the proliferation capacity of HBx and mutant were enhanced compared to the control group (P<0.01), HBx C1653T displayed further additive the effect compared to HBx (P<0.05). Moreover, HBxC1653T mutation also significantly upregulated N-cadherin expression and downregulated E-cadherin expression, thus promoting the occurrence of EMT. Conclusions　HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed, HBxC1653T mutation significantly enhanced the proliferation of HCC cells and epithelial to mesenchymal transition occurrence.

Prognostic value of age, creatinine, and left ventricular ejection fraction risk score in patients evaluated with fractional flow reserve: a cross-sectional study

SUMMARY OBJECTIVE: In this study, we investigated the relationship between age, creatinine, and left ventricular ejection fraction risk score and the severity of coronary lesions detected by applying fractional flow reserve in the patient group presenting with chronic coronary syndrome. Also, we presented long-term follow-up results in patients whose age, creatinine, and left ventricular ejection fraction score was evaluated by the fractional flow reserve procedure. METHODS: This study was planned retrospectively and in two centers. For this purpose, 114 patients who met the study criteria and who underwent elective fractional flow reserve between January 2014 and January 2019 were included in the study. Age, creatinine, and left ventricular ejection fraction was calculated as age/left ventricular ejection fraction +1 (if estimated glomerular filtration rate<30 mL/min). RESULTS: They were divided into two groups according to the cutoff value of the age, creatinine, and left ventricular ejection fraction score. A total of 76 patients had an age, creatinine, and left ventricular ejection fraction score of ≤1.17 (Group I) and 38 patients had an age, creatinine, and left ventricular ejection fraction score of >1.17 (Group II). The number of patients with severe lesions in fractional flow reserve was significantly higher in Group II compared with Group I (60.5 vs. 32.9%, p=0.005). According to the Kaplan-Meier analysis, a significant increase was observed in major adverse cardiac events and mortality during the follow-up period in the group with a high-risk score (Log Rank: 15.01, p<0.001 and Log Rank: 8.51, p=0.004, respectively). CONCLUSION: In light of the data we obtained from our study, we found a correlation between the severity of the lesion detected in fractional flow reserve and the age, creatinine, and left ventricular ejection fraction scores. In addition, we found that patients with high age, creatinine, and left ventricular ejection fraction scores had higher mortality and major adverse cardiac events rates during follow-up.

An increase in a long noncoding RNA ANRIL in peripheral plasma is an indicator of stable angina

Abstract Introduction: Stable angina develops during physical activity or stress, and it is typically an aspect of Coronary Heart Disease (CHD) that can lead to arrhythmia, heart failure and even sudden death. ANRIL, an Antisense Non-coding RNA gene in the INK4 Locus, is associated with multiple disorders including CHD; however, expressional levels of ANRIL in between patients with stable angina and myocardial infarction, one of the acute coronary syndrome, have not been clarified yet. Methods: The authors enrolled 62 patients with myocardial infarction and 59 with stable angina before primary percutaneous coronary intervention, as well as 48 healthy volunteers. Their peripheral blood was collected for analysis of ANRIL and cardiac troponin I, a traditional diagnostic index of CHD by real-time PCR. Results: The data showed that ANRIL is a better diagnostic indicator than cardiac troponin I in patients with stable angina and that the levels of ANRIL are higher in patients with stable angina than those with the myocardial infarction. Discussion: The levels of ANRIL in peripheral plasma could be used as a good biomarker for stable angina.

Chronic Coronary Syndrome: Medical Therapy or Myocardial Revascularization?

Abstract The best therapeutic strategy for chronic coronary syndrome (CCS) is still controversial. The lack of contemporaneity of medical treatment in many randomized clinical trials prior to the large-scale use of statins, antiplatelet agents, anti-diabetic drugs with cardiovascular protection, and changes in life habits with well-established goals limits the applicability of such studies in current clinical practice. Medical treatment is the only therapeutic option capable of reducing atherosclerotic damage and, therefore, of acting effectively in preventing the progression of this disease. The purpose of this brief review is to critically analyze the main contemporary studies that confront medical treatment with myocardial revascularization in CCS.

A survey of Stomoxys Geoffroy, 1762 (Diptera: Muscidae) in eight administrative regions of Cameroon

ABSTRACT Stomoxys Geoffroy, 1762 are major livestock pests in the tropics and are common in diverse habitats. This study aims to conduct a survey on the Stomoxys fauna of Cameroon. From 2015 to 2017, entomological studies using standard traps (n=204) were conducted in eight administrative regions found in five agro-ecological zones (AEZs) of Cameroon with 606 trap-points over 22,032 traps days. A total of 77,804 Stomoxys specimens were collected, with eight taxa consisting of six species (S. calcitrans (Linnaeus, 1758), S. omega Newstead, Dutton & Todd, 1907, S. xanthomelas Roubaud, 1937, S. inornatus Grunberg, 1906, S. transvittatus Villeneuve, 1916, and S. sitiens (Rondani, 1873)) and two subspecies (Stomoxys niger niger Macquart, 1851 and S. niger bilineatus Grunberg, 1906) identified. Among all the recorded taxa, S. calcitrans and S. n. niger were present in five and seven of the eight regions respectively, but S. sitiens was rare and only found in the North. Furthermore, the highest species number (seven out of eight) was recorded in the Guinee savanna of the Adamawa region. The highest apparent density range of 101 to 200 Stomoxys/ trap/ day (s/t/d) was recorded in the Sudan savanna AEZ of the Far North region. Stomoxys occurred in all the AEZs in eight regions, some of which are major cattle rearing regions. This represents risk for the mechanically transmission of dangerous pathogens in those regions.

Executive Summary of the Recommendations on the Management of Asthma In Primary Care (2022) – New Updates

Background : Asthma is a heterogenous disease defined by the history of respiratory symptoms (shortness of breath, wheezing, cough, and chest tightness) that vary over time and in intensity, along with variable expiratory airflow limitation. Despite an ever-increasing prevalence of asthma across all age groups, this condition remains poorly managed in India. Majority of the Indian patients remain undiagnosed or wrongly diagnosed in general clinical practice and even those who get diagnosed, remain poorly or inadequately treated1,2. Since the last published 2020 Indian Medical Association (IMA) recommendations on the management of asthma in primary care, noteworthy critical changes have been recommended in relation to the diagnosis/management of asthma in international guidelines. Hence, there was a need to update the existing IMA recommendations. For the same, an expert group meeting was organized with family physicians having clinical experience in managing patients with asthma along with chest physicians and pediatricians. Important updates related to asthma diagnosis and its management were discussed and the final recommendation decisions were derived from the joint group discussion. Some of the key points derived from the discussion are mentioned below in the executive summary. For a detailed version of the new recommendations please click on the url.