Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90％-110％.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all＜5％.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.

Objective:To explore the effects of coenzyme Q10(CoQ10) on high-fat diet-induced obesity, lipid disorders, and bile acid metabolism in mice.Methods:Eight-week-old C57BL/6J mice were randomly divided into control group(regular chow), high-fat diet group(45% high-fat chow), and CoQ10 intervention group(45% high-fat chow+ 100 mg·kg -1·d -1CoQ10) based on their body weights according to the randomized block design. The body weight and food intake of mice in each group were collected. The levels of serum total cholesterol, triglyceride, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, alanine aminotransferase, and aspartate aminotransferase were detected. The contents of 17 bile acids in serum, liver, and colon contents of mice were detected by ultra-performance liquid chromatography-tandem mass(UPLC-MS/MS). The protein expressions of cholesterol 12α-hydroxylase(CYP8B1) and oxysterol 7α-hydroxylase(CYP7B1) in liver were detected by Western blotting. Results:CoQ10 significantly reduced body weight and ameliorated lipid metabolism disorders in mice fed a high-fat diet. Compared with the control group, serum total bile acid levels were reduced in the high-fat diet group( P<0.05); CoQ10 intervention elevated serum and colonic total bile acid levels( P=0.021, P=0.014) and increased liver, colon, and serum deoxycholic acid and ursodeoxycholic acid levels( P<0.05) in the mice compared with the high-fat diet group. Both colonic and serum deoxycholic acid levels in the CoQ10 intervention group were negatively correlated with body weights( P=0.024, P=0.019), and colonic deoxycholic acid and total cholesterol levels were also negatively correlated( P=0.006). CoQ10 increased the expression of CYP8B1 and CYP7B1 proteins in the liver of mice. Conclusion:CoQ10 can modulate bile acid metabolism in high-fat diet-fed mice and alleviate their obesity and lipid metabolism disorders.

Objective:To analyze the classification and influencing factors of post-intensive care syndrome in family members of critically ill children, so as to provide reference for developing targeted intervention strategies.Methods:Using cross-sectional survey methods, the family members of critically ill children from January 2022 to June 2023 in the First Affiliated Hospital of Sun Yat-sen University were selected as research objects by convenience sampling method. The general information questionnaire, Pittsburgh Sleep Quality Index(PSQI), Fatigue Assessment Instrument(FAI), Hospital Anxiety and Depression Scale(HADS), and Impact of Event Scale-Revised(IES-R) were used for investigation, respectively.Results:Finally, 131 family members of critically ill children were included in the present study, 37 males, 94 females, aged 28-67 (39.47 ± 8.55) years. The incidence of PICS-F were 77.9% (102/131), the scores of PSQI, FAI, anxiety, depression and IES-R were (7.95 ± 3.39), (3.42 ± 1.34), (14.02 ± 5.47), (14.10 ± 5.54), (39.14 ± 7.92) points, respectively. The types of PICS-F in respondents were divided into four subtypes: sleep disturbance type were 42 cases accounting for 32.1%, stress disturbance type were 17 cases accounting for 13.0%, PICS-F high risk type were 40 cases accounting for 30.5%, PICS-F low risk type were 32 cases accounting for 24.4%. The length of ICU hospitalization was a common influencing factor for sleep disturbance type ( OR=0.103, 95% CI 0.020-0.520), stress disturbance type ( OR=0.073,95% CI 0.011-0.467), and PICS-F high risk type ( OR=0.115, 95% CI 0.022-0.589), all P<0.05. The per monthly income was a common risk factor for sleep disturbance type ( OR=5.510, 95% CI 1.193-5.445) and PICS-F high risk type ( OR=9.302, 95% CI 1.435-2.306), both P<0.05; the relationship with children was a specific risk factor for sleep disturbance type ( OR=0.083, 95% CI 0.009-0.734, P<0.05); the APACHEⅡscore was a specific risk factor for PICS-F high risk type ( OR=0.091, 95% CI 0.014-0.586, P<0.05). Conclusions:There are four latent profiles of PICS-F in family members of critically ill children, medical staff should establish targeted interventions according to the characteristics of different PICS-F classes.

Gynecologic malignant tumors are among the leading diseases threatening women's lives and health,with the highest morbidity and mortality rates among all female diseases.These tumors originate from female reproductive organs and are typically classified based on the affected site.Ovarian cancer(OC),endometrial cancer(EC)and cervical cancer(CCA)are the most common types.Currently,gynecologic malignant tumors are primarily treated with a combination of surgery,chemotherapy and radiotherapy,where drugs play a critical role in the treatment process.However,the actual clinical effectiveness is often influenced by various factors,such as adverse reactions due to drug toxicity and the drug resistance and insensitivity observed in some patients,which limit improvements in patient survival rates.Recent studies have shown that the same type of tumor exhibits significant biological characteristics and drug response heterogeneity among different individuals,which is a key factor contributing to the varied clinical outcomes when using the same drug treatment for the same type of gynecologic malignant tumor.To achieve individualized and precise treatment for gynecologic malignant tumors,there is an urgent need to develop in vitro models that closely resemble human tumors for clinical research.Drug screening is a technique used to identify and evaluate compounds with pharmacological activity and potential therapeutic effects,providing doctors with scientific guidance on drug use,thereby avoiding blind drug testing and reducing patients'therapeutic pain and economic burden by assessing the effects of different drugs under specific conditions.Organoid models have been extensively studied as an innovative drug screening tool and personalized medicine for treating gynecologic malignancies.Organoids are tissue-like structures with a specific spatial arrangement formed in vitro through three-dimensional cell culture,capable of highly simulating the structure and function of tissues in vivo and displaying histological and genotypic characteristics very similar to human organs.This approach has largely overcome the limitations of traditional tumor models,such as patient-derived cancer cell models and patient-derived tumor xenograft models,becoming an essential research tool in oncology.It provides a more physiologically relevant experimental platform for drug screening studies of gynecologic malignancies.This paper compared the advantages and disadvantages of several preclinical cancer models,reviewed the development process of organoids,and described the establishment of gynecologic oncology organoids and their application in drug screening for ovarian,endometrial,and cervical cancers.Additionally,we discussed the current limitations of organoid technology in its application and envisioned its future development,aiming to provide insights for future medical research,particularly in new drug discovery and personalized medicine.

Objective To investigate the correlation between serum osteoblastin(OPN)and angiopoietin-like protein 8(ANGPTL8)levels and hepatic fibrosis(HF)after interventional therapy-transcatheter arterial chemoembolisation(TACE)in patients with primary hepatocellular carcinoma(PHC).Methods 166 patients with PHC admitted between March 2021 and June 2023 were selected and divided into 92 cases with HF(observation group)and 74 cases without HF(control group)according to whether or not HF occurred after interventional therapy;enzyme-linked immunosorbent assay(ELISA)was used to determine the serum OPN and ANGPTL8 levels and to analyse the predictive value of the OPN and ANGPTL8 levels on HF.Pearson correlation was used to analyze the correlation between OPN and ANGPTL8 levels and biochemical indexes.The factors influencing the occurrence of HF were analyzed by multi-factor Logistics regression.ROC curve was used to analyze the predictive value of OPN and ANGPTL8 for HF.Results Serum OPN[(74.56±11.56)ng/ml],ANGPTL[(42.78±5.23)ng/ml],ALT[(62.24±9.56)U/L],AST[(42.88±8.23)U/L],HA[(252.98±52.44)ng/L],LN[(152.64±26.45)ng/L],PC Ⅲ[(16.54±3.46)ng/L]and Ⅳ-C[(152.78±21.23)ng/L]in observation group were significantly higher than the control group[(57.89±9.68)ng/ml,(35.46±4.78)ng/ml,(49.46±7.46)U/L,(31.48±7.26)U/L,(192.56±23.88)ng/L,(124.48±11.23)ng/L,(11.26±2.23)ng/L and(126.45±18.56)ng/L].The differences between the two groups were statistically significant(P＜0.05).The AUC of serum OPN,ANGPTL8 and the combination of the two in predicting the occurrence of HF were 0.914,0.920 and 0.978,respectively,and the AUC of OPN combined with ANGPTL8 in predicting the occurrence of HF was higher than the AUC of the two separately(P＜0.05).Conclusion The levels of serum OPN and ANGPTL8 of patients with PHC are closely associated with the occurrence of HF,and the two are HF occurrence influencing factors and can be used as indicators to predict the occurrence of HF.

Objective:To investigate the expression level of small nucleolar RNA SNORD15A in bone marrow of patients with acute leukemia (AL) and its relationship with clinical characteristics and prognosis of patients.Methods:Bone marrow blood samples of 53 newly treated AL patients and 29 healthy subjects without clinical diagnosis of hematologic diseases or other malignant diseases (control group) at the Affiliated Hospital of Guangdong Medical University from March 2018 to December 2021 were collected. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression of SNORD15A in bone marrow blood mononuclear cells of the two groups. The median relative expression of SNORD15A (0.148) was used as the boundary, and AL patients were divided into low expression group (<0.148) and high expression group (≥0.148). The relationship between the expression level of SNORD15A and the clinical characteristics, clinical indicators and overall survival (OS) of AL patients was analyzed. Kaplan-Meier method was used for survival analysis and log-rank test was performed; Cox proportional hazards model was used for univariate and multivariate analyses of OS of patients.Results:The relative expression of SNORD15A was 0.148 (0.012-1.376) in newly treated AL patients and 0.921 (0.513-2.288) in the control group, and the difference was statistically significant ( Z = -6.85, P < 0.01). The differences in SNORD15A relative expression between patients with different prognostic stratification, efficacy and with or without fever and bleeding were statistically significant (all P < 0.05). The differences in platelet count, plateletcrit and albumin levels between SNORD15A low expression group and high expression group were statistically significant (all P < 0.05), and the differences in molecular biology and cytogenetic characteristics were not statistically significant (all P > 0.05). The patients in SNORD15A high expression group had better OS than the low expression group ( P < 0.05). The results of univariate Cox regression analysis showed that SNORD15A was an influencing factor for patients' OS ( HR = 0.063, 95% CI 0.005-0.766, P < 0.05); the results of multivariate Cox regression analysis showed that fatigue ( HR = 4.754, 95% CI 1.014-22.290), fever ( HR = 0.147, 95% CI 0.029-0.746) and hemoglobin ( HR = 0.970, 95% CI 0.944 -0.998) were independent influencing factors for OS (all P < 0.05). Conclusions:SNORD15A is lowly expressed in AL and may be an indicator for disease monitoring and prognostic assessment in AL patients.

Objective:To explore the clinical effect and safety of interventional treatment of autogenous arteriovenous fistula(AVF) stenosis.Methods:From July 2017 to September 2018, 96 patients with arteriovenous fistula stenosis and occlusion admitted to Handan First Hospital, Hebei Province were retrospectively analyzed.All of them were dialysis patients with chronic renal failure.All patients underwent percutaneous balloon angioplasty via the cephalic vein.The success rate of technique, clinical success rate, perioperative complications and follow-up were observed.Results:(1) Technical success rate and clinical success rate: 90 patients were treated with percutaneous transluminal angioplasty (PTA) via the cephalic vein, the other 3 patients were treated with interventional therapy via the brachial artery, and 3 patients underwent reconstruction of internal fistula.The technical success rate was 93.8% (90/96), and the clinical success rate was 89.6% (86/96). (2) Perioperative complications: thrombosis in 4 cases, vasospasm in 3 cases.There were no serious complications such as vascular rupture, aneurysm, vascular dissection, and no perioperative death.(3) The first stage patency rate was 100% (90/90), 74.4% (67/90), 62.2% (56/90) and 46.7% (42/90) in 3, 6, 12 and 18 months after operation.Conclusion:Venipuncture can be used as the first choice for AVF stenosis interventional therapy because of its advantages of small trauma, no serious complications, no need of long-term compression at the puncture point, immediate dialysis, and avoidance of local hematoma and other complications caused by artery puncture.

Objective:To compare the clinical effect and perioperative complications of the treatment of autogenous arteriovenous fistula stenosis by arterial and venous approach.Methods:The clinical data of 120 patients with AVF stenosis and occlusion who were treated with interventional therapy and met the inclusion criteria were collected and analyzed by retrospective case-control study.from September 2017 to August 2018, 60 patients with internal fistula stenosis were treated by transarterial approach (arterial approach group), and from September 2018 to may 2019, 60 patients were treated with a new surgical scheme(venous approach group). The operation success rate, perioperative complications and patency rate of 3, 6, 12 months after operation were compared between the two groups.Results:(1) The technical success rate was 96.7% (58/60) and the clinical success rate was 91.7% (55/60) in the arterial approach group, and 95.0%(57/60) and 93.3%(56/60) in the venous approach group.There was no significant difference in the technical success rate and clinical success rate between the two groups ( P=0.718 and 1.000, respectively) (2) Perioperative complications: in the arterial approach group, 3 patients had hematoma at the puncture point, 2 pseudoaneurysms and 5 thrombosis.There were 3 patients with thrombosis in the venous access group, and the difference in the incidence of complications between the two groups was statistically significant (χ 2=4.227, P=0.036). (3)The primary patency rates at 3, 6 and 12 months after operation were 95.0%(57/60), 75.0%(45/60) and 60.0%(36/60) in the arterial approach group, and 96.7%(58/60), 71.7%(43/60) and 61.7%(37/60) in the venous access group, respectively.There was no statistically significant difference between the two groups ( P=0.718, 0.749, 0.885). Conclusion:The interventional treatment for autogenous arteriovenous fistula stenosis through artery and vein approach can achieve good effect.There were many complications during the perioperative period, It is suggested that venous approach is preferred.

Objective:To investigate the pathological characteristics and the clinical significance of novel coronavirus (2019-nCoV)-infected pneumonia (termed by WHO as coronavirus disease 2019, COVID-19).Methods:Minimally invasive autopsies from lung, heart, kidney, spleen, bone marrow, liver, pancreas, stomach, intestine, thyroid and skin were performed on three patients died of novel coronavirus pneumonia in Chongqing, China. Hematoxylin and eosin staining (HE), transmission electron microcopy, and histochemical staining were performed to investigate the pathological changes of indicated organs or tissues. Immunohistochemical staining was conducted to evaluate the infiltration of immune cells as well as the expression of 2019-nCoV proteins. Real time PCR was carried out to detect the RNA of 2019-nCoV.Results:Various damages were observed in the alveolar structure, with minor serous exudation and fibrin exudation. Hyaline membrane formation was observed in some alveoli. The infiltrated immune cells in alveoli were majorly macrophages and monocytes. Moderate multinucleated giant cells, minimal lymphocytes, eosinophils and neutrophils were also observed. Most of infiltrated lymphocytes were CD4-positive T cells. Significant proliferation of type Ⅱ alveolar epithelia and focal desquamation of alveolar epithelia were also indicated. The blood vessels of alveolar septum were congested, edematous and widened, with modest infiltration of monocytes and lymphocytes. Hyaline thrombi were found in a minority of microvessels. Focal hemorrhage in lung tissue, organization of exudates in some alveolar cavities, and pulmonary interstitial fibrosis were observed. Part of the bronchial epithelia were exfoliated. Coronavirus particles in bronchial mucosal epithelia and type Ⅱ alveolar epithelia were observed under electron microscope. Immunohistochemical staining showed that part of the alveolar epithelia and macrophages were positive for 2019-nCoV antigen. Real time PCR analyses identified positive signals for 2019-nCoV nucleic acid. Decreased numbers of lymphocyte, cell degeneration and necrosis were observed in spleen. Furthermore, degeneration and necrosis of parenchymal cells, formation of hyaline thrombus in small vessels, and pathological changes of chronic diseases were observed in other organs and tissues, while no evidence of coronavirus infection was observed in these organs.Conclusions:The lungs from novel coronavirus pneumonia patients manifest significant pathological lesions, including the alveolar exudative inflammation and interstitial inflammation, alveolar epithelium proliferation and hyaline membrane formation. While the 2019-nCoV is mainly distributed in lung, the infection also involves in the damages of heart, vessels, liver, kidney and other organs. Further studies are warranted to investigate the mechanism underlying pathological changes of this disease.

The main components of Schisandra chinensis are γ-schizandrin, schisandrin A,deoxyschizandrin,schisandra B, schisandrol B and so on. Schisandra contains a variety of bioactive ingredients mainly used in clinics and combined with other drugs. Many of the drugs currently used in clinical practice can cause drug-induced liver injury(DILI),which is one of the main adverse drug reactions,therefore,the clinical use of drugs for the prevention and treatment of liver damage is particularly important. In this paper, the effective components of Schisandra chinensis and their related therapeutic effects on DILI were reviewed.