Virus load and virus shedding of SARS-CoV-2 and their impact onpatient outcomes

BACKGROUND Understanding a virus shedding patterns in body fluids/secretions is importantto determine the samples to be used for diagnosis and to formulate infectioncontrol measures.AIM To investigate the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)shedding patterns and its risk factors.METHODS All laboratory-confirmed coronavirus disease 2019 patients with completemedical records admitted to the Shenzhen Third People’s Hospital from January28, 2020 to March 8, 2020 were included. Among 145 patients (54.5% males;median age, 46.1 years), three (2.1%) died. The bronco-alveolar lavage fluid(BALF) had the highest virus load compared with the other samples. The viralload peaked at admission (3.3 × 108 copies) and sharply decreased 10 d afteradmission.RESULTS The viral load was associated with prolonged intensive care unit (ICU) duration.Patients in the ICU had significantly longer shedding time compared to those inthe wards (P < 0.0001). Age > 60 years [hazard ratio (HR) = 0.6;95% confidenceinterval (CI): 0.4-0.9] was an independent risk factor for SARS-CoV-2 shedding,while chloroquine (HR = 22.8;95%CI: 2.3-224.6) was a protective factor.CONCLUSION BALF had the highest SARS-CoV-2 load. Elderly patients had higher virus loads,which was associated with a prolonged ICU stay. Chloroquine was associatedwith shorter shedding duration and increased the chance of viral negativity.

Transforaminal Endoscope for the Treatment of Thoracic Disc Herniation

Objective:Transforaminal endoscope was used in the treatment of thoracic disc herniation and the therapeutic effect was discussed.Methods:Sixteen patients with thoracic disc herniation admitted to our hospital from October 2018 to May 2019 were divided into control group and observation group according to the random number table method,with 8 patients in each group.The control group was treated with posterior laminectomy approach,and the observation group was treated with transforaminal endoscope.The surgery-related conditions(intraoperative blood loss,surgery time,and postoperative drainage volume),VAS score,JOA score and postoperative complications were compared and analyzed.Results:There was no statistically significant difference between the two groups in the total effective rate and preoperative VAS score(P>0.05).Compared with the control group,the observation group had shorter surgery time and incision length,less postoperative drainage volume,and higher VAS score 3 days after surgery,JOA score on day 7 after surgery,and JOA score 3 months after surgery,showing statistically significant differences(P<0.05).There was no significant difference in the complications between the two groups(X2=1.067,P=0.833).Conclusions:Transforaminal endoscope had a good clinical effect with small incision and injury in the treatment of patients with thoracic disc herniation.

Preliminary study of transforaminal endoscope in the treatment of spinal metastasis

Objective:To investigate the therapeutic effect of transforaminal endoscope in patients with spinal metastases.Methods:Clinical data of 28 patients with spinal metastases admitted to our hospital from May 2015 to May 2019 were retrospectively collected.According to different surgical methods,they were divided into control group(20 cases)and experimental group(8 cases),among which control group was treated with traditional open palliative decompression,while the experimental group was treated with transforaminal endoscopic decompression.The surgery and recovery indicators were compared between the two groups,including surgery time,incision length,postoperative drainage volume,out of bed activity time,length of hospital stay,the Japanese Orthopaedic Association(JOA)scores of 7 days after surgery,pain degree,complications(wound infection,transient reduction of muscle strength,hypoproteinemia with wound drainage and delayed healing),and activity of daily living.Results:The surgery time,incision length,postoperative drainage volume,out of bed activity time,and postoperative length of hospital stay of the experimental group were all less than those of the control group,and the JOA score of 7 days after surgery was higher than that of the control group,showing statistically significant differences(P<0.05).VAS scores of the experimental group on the day 1 and day 7 after surgery were lower than those of the control group,with statistically significant differences(P<0.05).The incidence of complications in the experimental group was slightly lower than that in the control group,but the difference was not significant(P>0.05).Conclusions:Transforaminal endoscope used in suitable patients with spinal metastases can greatly reduce the incision length,soft tissue and bone tissue damages,and postoperative drainage,promote early mobilization and early discharge,and reduce a series of complications due to hemorrhage and hypoproteinemia,which has a better early clinical effect in comparison with the traditional open palliative decompression.

Screening of differentially expressed microRNA in ulcerative colitis related colorectal cancer

Objective:To investigate the differential expression of microRNA(miRNA)in colon between ulcerative colitis(UC)and ulcerative colitis related colorectal cancer(UCRCC).Methods:An UC mouse model was built by dextran sodium sulfate,and an UCRCC mouse model by dextran sodium sulfate and 1,2-diformylhydrazine.RNAs were extracted from the colon,purified and hybridized with fluorescence-labeled miRNA oligonucleotide gene chip.Real-time fluorescence quantitative PCR was used to verity the expression variation of miRNA.SAM was employed for the data analysis.Results:The up-regulated miRNA in colon cancer included has-miR-194,hasmiR-215,has-miR-93,has-miR-192,has-miR-92a,has-miR-29b,and has-miR-20a(median false discovery rate<5%),while the down-regulated miRNAs were has-miR-1231,has-miR-195,has-miR-143,and has-miR-145(median false discovery rate<5%).Conclusions:Significant differential expression of miRNA was found between the UC mouse and UCRCC mouse,which may be related to the onset,erosion and transfer of colorectal cancer.

Effects of glycemic control on refraction in diabetic patients

AIMTo evaluate the effects of glycemic control on refraction in diabetic patients.

Clinical diagnosis of severe COVID-19:A derivation and validation of a prediction rule

BACKGROUND The widespread coronavirus disease 2019(COVID-19)has led to high morbidity and mortality.Therefore,early risk identification of critically ill patients remains crucial.AIM To develop predictive rules at the time of admission to identify COVID-19 patients who might require intensive care unit(ICU)care.METHODS This retrospective study included a total of 361 patients with confirmed COVID-19 by reverse transcription-polymerase chain reaction between January 19,2020,and March 14,2020 in Shenzhen Third People’s Hospital.Multivariate logistic regression was applied to develop the predictive model.The performance of the predictive model was externally validated and evaluated based on a dataset involving 126 patients from the Wuhan Asia General Hospital between December 2019 and March 2020,by area under the receiver operating curve(AUROC),goodness-of-fit and the performance matrix including the sensitivity,specificity,and precision.A nomogram was also used to visualize the model.RESULTS Among the patients in the derivation and validation datasets,38 and 9 participants(10.5%and 2.54%,respectively)developed severe COVID-19,respectively.In univariate analysis,21 parameters such as age,sex(male),smoker,body mass index(BMI),time from onset to admission(>5 d),asthenia,dry cough,expectoration,shortness of breath,asthenia,and Rox index<18(pulse oxygen saturation,SpO2)/(FiO2×respiratory rate,RR)showed positive correlations with severe COVID-19.In multivariate logistic regression analysis,only six parameters including BMI[odds ratio(OR)3.939;95%confidence interval(CI):1.409-11.015;P=0.009],time from onset to admission(≥5 d)(OR 7.107;95%CI:1.449-34.849;P=0.016),fever(OR 6.794;95%CI:1.401-32.951;P=0.017),Charlson index(OR 2.917;95%CI:1.279-6.654;P=0.011),PaO2/FiO2 ratio(OR 17.570;95%CI:1.117-276.383;P=0.041),and neutrophil/lymphocyte ratio(OR 3.574;95%CI:1.048-12.191;P=0.042)were found to be independent predictors of COVID-19.These factors were found to be significant risk factors for severe patients confirmed with COVID-19.The AUROC was 0.941(95%CI:0.901-0.981)and 0.936(95%CI:0.886-0.987)in both datasets.The calibration properties were good.CONCLUSION The proposed predictive model had great potential in severity prediction of COVID-19 in the ICU.It assisted the ICU clinicians in making timely decisions for the target population.

Clinical Application of PES Membrane Dialyzer

Objective The dialysate membrane can be classified into acetate membrane and synthetic membrane,while the latter can be further divided into polysulfone,polyacrylonitrile,etc.Polyethersulfone membrane is high molecular membrane material with excellent biocompatibility,stable mechanical properties,high temperature and pressure resistance,and good cleaning performance,which has been applied for more than 10 years.This trial is to assess the safety and efficacy of hollow polysulfone membrane fiber dialyzer with pattern number Enttex^(TM)-16LF(E60)for hemodialysis in patients with acute or chronic renal failure due to multiple factors.Methods Totally 76 patients receiving maintenance hemodialysis were included in a two-site,random,open label,two-stage,positive crossover and non inferiority validation study.The clinical efficacy and safety of the test device were observed,which was a hollow fiber dialyzer produced by Guangzhou Enttex Medical Products Co.,Ltd.,with polyethersulfone membrane imported from Germany.It was a low flux dialyzer with the pattern number of EnttexTM-16LF(E60).Inclusion criteria:①patients with acute or chronic renal failure due to various reasons who needed hemodialysis;②18 to 80 years old,gender unlimited;③patients who received stable dialysis for more than 3 months,regular hemodialysis 3 times a week,4 h at each time,blood flow rate of 200~350ml/min;④using bicarbonate dialysate at a flow rate of 500ml/min;⑤patients with arteriovenous fistula or who received artificial blood vessel puncture dialysis;received anticoagulation by heparin or low molecular weight heparin.Exclusion criteria:①patients needing heparin free dialysis for severe anemia,tumor and active bleeding;②patients with severe cardiac(gradeⅢcardiac function),hepatic(severe hepatitis and cirrhosis)and pulmonary diseases(severe respiratory failure).Results Comparison of parameters,including the changes of Kt/V,general creatinine and urea clearance rates and URR(urea reduction rate)shows no statistical significance in differences between the two devices,indicating that the test and control devices can equally and effectively eliminate toxins including creatinine and urea,etc.PPP and MITTP results are consistent.Comparison of ultrafiltration rates between two groups of patients shows no statistical difference.PPP and MITTP results are consistent,indicating that both the test device and control device can effectively eliminate water.In MITTP,electrolytes including K^+,Na^+,Cl^-,Ca^2+,HCO3^-,etc.show no statistical significance in difference between the test and control devices before and after dialysis(P>0.05).After treating by using test and control devices,body weight,serum potassium and phos-phorus all significantly decrease(P<0.001);serum Ca^2+,PO2,PCO2and HCO3^-all significantly increase(P<0.001).These results indicate that both the test and control devices can effectively eliminate serum potassium and phosphorus,and eliminate excess water in body,maintain the balance between liquid and electrolytes in patients.PPP and MITTP results are consistent.Conclusion This clinical trial confirms that hollow polyethersulfone membrane fiber dialyzer can be safely and effectively used in hemodialysis therapy.

Successful management of seven cases of critical COVID-19 with early noninvasive-invasive sequential ventilation algorithm and bundle pharmacotherapy

We report the clinical and laboratory findings and successful management of seven patients with critical coronavirus disease 2019(COVID-19)requiring mechanical ventilation(MV).The patients were diagnosed based on epidemiological history,clinical manifestations,and nucleic acid testing.Upon diagnosis with COVID-19 of critical severity,the patients were admitted to the intensive care unit,where they received early noninvasive-invasive sequential ventilation,early prone positioning,and bundle pharmacotherapy regimen,which consists of antiviral,anti-inflammation,immune-enhancing,and complication-prophylaxis medicines.The patients presented fever(n=7,100%),dry cough(n=3,42.9%),weakness(n=2,28.6%),chest tightness(n=1,14.3%),and/or muscle pain(n=1,14.3%).All patients had normal or lower than normal white blood cell count/lymphocyte count,and chest computed tomography scans showed bilateral patchy shadows or ground glass opacity in the lungs.Nucleic acid testing confirmed COVID-19 in all seven patients.The median MV duration and intensive care unit stay were 9.9 days(interquartile range,6.5-14.6 days;range,5-17 days)and 12.9 days(interquartile range,9.7-17.6 days;range,7-19 days),respectively.All seven patients were extubated,weaned off MV,transferred to the common ward,and discharged as of the writing of this report.Thus,we concluded that good outcomes for patients with critical COVID-19 can be achieved with early noninvasive-invasive sequential ventilation and bundle pharmacotherapy.

Total Synthesis and Biological Evaluation of Wewakazole

The total synthesis of Wewakazole was achieved on a gram scale, unambiguously confirming its structure. Three different substituted oxazoles present in this natural product were prepared efficiently. Biological evaluation indicated that Wewakazole significantly inhibited cancer cell proliferation in a dose-dependent manner.

腹腔镜意外胆囊癌外科治疗体会

目的总结意外胆囊癌腹腔镜胆囊切除的经验。方法回顾性分析连续1037例腹腔镜胆囊切除术中的4例意外胆囊癌患者的临床特征、病理分期、手术及术后处理措施。结果术后病理分期2例为T1aNM,1例为00T1bNM,1例为TisNM,其中3例肿瘤位于体部,1例位于胆囊颈部息肉癌变。术后随访1～3年,无切口种植,腹腔0000及远处转移。结论LC只要注意技术操作,勿分破胆囊,并不会增加肿瘤扩散的机会;术后应根据PTNM分期及肿瘤部位决定是否行淋巴清扫。

Inhibition of squamous cancer growth in a mouse model by Staphylococcal enterotoxin B-triggered Th9 cell expansion

Currently, therapy for squamous cancer （SqC） is unsatisfactory. Staphylococcal enterotoxin B （SEB） has strong immune regulatory activity. This study tests the hypothesis that SEB enforces the effect of immunotherapy on SqC growth in a mouse model. C3H/HeN mice and the SqC cell line squamous cell carcinoma VII were used to create an SqC mouse model. Immune cell assessment was performed by flow cytometry. Real-time RT-PCR and western blotting were used to evaluate target molecule expression. An apoptosis assay was used to assess the suppressive effect of T helper-9 （Th9） cells on the SqC cells. The results showed that immunotherapy consisting of SEB plus SqC antigen significantly inhibited SqC growth in the mice. The frequency of Th9 cells was markedly increased in the SqC tissue and mouse spleens after treatment. SEB markedly increased the levels of signal transducer and activator of transcription 5 phosphorylation and the expression of histone deacetylase-1 （HDAC1） and PU.1 （the transcription factor of the interleukin 9 （IL-9） gene） in CD4^＋ T cells. Exposure to SqC-specific Th9 cells markedly induced SqC cell apoptosis both in vitro and in vivo. In conclusion, the administration of SEB induces Th9 cells in SqC-bearing mice, and theseTh9 cells inhibit SqC growth.

Influence of lesion ratio on diagnostic performance of in-phase/opposed-phase imaging and apparent diffusion coefficient for differentiating acute benign vertebral fractures and metastases

Background The usefulness of in-phase/opposed-phase imaging and diffusion weighted imaging （DWI） in differentiating benign and neoplastic vertebral fractures has been described. In this study, we aimed to evaluate the influence of the severity of vertebral damage on the diagnostic performance of these two technologies. Methods Totally 59 patients with 68 acute benign vertebral fractures and 43 patients with 79 vertebral metastases were included in this study. The MR protocol included DWls and sagittal in-phase/opposed-phase gradient recalled sequence. The severity of vertebral damage was expressed by lesion ratio （LR, the ratio of lesion area to vertebral area on the slices of largest abnormal signal area in the Tl-weighted sequence）. Quantitative （signal intensity ratio （SIR） defined as signal intensity （SI） on opposed-phase gradient recalled echo （GRE） images divided by SI on in-phase; apparent diffusion coefficient （ADC） value derived from DWl analysis was performed, the relationships between LR and the measurements of these two technologies were analyzed using linear regression. The covariate-specific receiver operating characteristic （ROC） curves were also fitted to evaluate the influence of LR on the diagnostic performance of ADC and SIR. Results The difference in both SIR and ADC for vertebral metastasis and acute benign vertebral fractures was significant （P 〈0.001）. A positive correlation between the LR and the SIR was found in benign fractures （P 〈0.05）. The severity of vertebral damage had a significant influence on the AUC （area under ROC curve） for SIR （P 〈0.05） but ADC （P 〉0.05）. More severe cases were associated with increased AUC for SIR. Conclusions LR is capable of affecting the diagnostic performances of chemical shift imaging. Thus, when applying these tests to make diagnoses on vertebral fractures, the severity of the vertebral damage should be taken into account. The covariate-specific ROC model is recommended because it substantially improves the ability to avoid bias when evaluating tests.

Neuroprotective Effect of Fructus broussonetiae on APP/PS1 Mice via Upregulation of AKT/β-Catenin Signaling

Objective:To evaluate the mechanisms underlying the protective effect of Chinese herbal medicine Fructus broussonetiae(FB)in both mouse and cell models of Alzheimer’s disease(AD).Methods:APP/PS1 mice treated with FB for 2 months and vehicle-treated controls were run through the Morris water maze and object recognition test to evaluate learning and memory capacity.RNA-Seq,Western blotting,and immunofluorescence staining were also conducted to evaluate the effects of FB treatment on various signaling pathways altered in APP/PS1 mice.To further explore the mechanisms underlying FB’s protective effect,PC-12 cells were treated with Aβ25–35 in order to establish an in vitro model of AD.Results:FB-treated mice showed improved learning and memory capacity on both the Morris water maze and object recognition tests.RNA-seq of hippocampal tissue from APP/PS1 mice showed that FB had effects on multiple signaling pathways,specifically decreasing cell apoptotic signaling and increasing AKT and β-catenin signaling.Similarly,FB up-regulated both AKT and β-catenin signaling in PC-12 cells pre-treated with Aβ25–35,in which AKT positively regulated β-catenin signaling.Further study showed that AKT promoted β-catenin signaling via enhancing β-catenin(Ser552)phosphorylation.Moreover,AKT and β-catenin signaling inhibition both resulted in the attenuated survival of FB-treated cells,indicating the AKT/β-catenin signaling is a crucial mediator in FB promoted cell survival.Conclusions:FB exerted neuroprotective effects on hippocampal cells of APP/PS1 mice,as well as improved cell viability in an in vitro model of AD.The protective actions of FB occurred via the upregulation of AKT/β-catenin signaling.