Objective To explore the possibility and genetic identification method of constructing a hepatocyte-specific NLRP3 gene knockout mouse model by using Cre-LoxP system gene knockout technology.Methods Phase one:mice specifically expressing the albumin promoter-Cre(AlbCre)recombinase in hepatocytes were mated with NLRP3flox/flox mice,and the hepatocyte-specific NLRP3 gene knockout mice with the genotype of NLRP3flox/flox/AlbCre+/-(hepatocyte NLRP3 knockout group)and the control mice in the same litter with the genotype of NLRP3flox/flox/AlbCre-/-(control group in the same litter)were obtained after two generations of selection and mating.The second stage was the mass reproduction stage.Mating NLRP3flox/flox/AlbCre+/-target mice with NLRP3flox/flox mice could quickly obtain a large number of experimental target mice and control mice in the same litter.The DNA was extracted from the tails of mice after numbering,and the offspring genotype was identified by PCR.qPCR and Western blot were used to detect the mRNA and protein expression levels of NLRP3 gene in the liver tissue.HE staining was used to observe the morphological changes in liver tissues,and serum liver transaminases and inflammatory factors were detected.The changes in body weight,liver-to-body ratio and special circumstances during reproduction and development of mice in the two groups were observed.Results The offspring genotype of the target mice in the F2 generation was consistent with theoretical result of NLRP3flox/flox/AlbCre+/-.The mRNA and protein levels of NLRP3 in liver tissues of mice in the hepatocyte NLRP3 knockout group were significantly lower than those in the control group in the same litter(P<0.05).The mice in the hepatocyte NLRP3 knockout group was not affected in terms of growth,development and reproduction after the NLRP3 gene knockout.There were no statistically significant differences in the body weight,liver-to-body ratio,liver tissue morphology,serum liver transaminase or inflammatory factors between the hepatocyte NLRP3 knockout group and the control group in the same litter(P>0.05).Conclusion The Cre-LoxP gene knockout technology can be used to successfully construct a hepatocyte-specific NLRP3 gene knockout mouse model,providing an important technical support for the next step of studying the function of the NLRP3 gene in the liver at the animal level.

Objective To explore the possibility and genetic identification method of constructing a hepatocyte-specific NLRP3 gene knockout mouse model by using Cre-LoxP system gene knockout technology.Methods Phase one:mice specifically expressing the albumin promoter-Cre(AlbCre)recombinase in hepatocytes were mated with NLRP3flox/flox mice,and the hepatocyte-specific NLRP3 gene knockout mice with the genotype of NLRP3flox/flox/AlbCre+/-(hepatocyte NLRP3 knockout group)and the control mice in the same litter with the genotype of NLRP3flox/flox/AlbCre-/-(control group in the same litter)were obtained after two generations of selection and mating.The second stage was the mass reproduction stage.Mating NLRP3flox/flox/AlbCre+/-target mice with NLRP3flox/flox mice could quickly obtain a large number of experimental target mice and control mice in the same litter.The DNA was extracted from the tails of mice after numbering,and the offspring genotype was identified by PCR.qPCR and Western blot were used to detect the mRNA and protein expression levels of NLRP3 gene in the liver tissue.HE staining was used to observe the morphological changes in liver tissues,and serum liver transaminases and inflammatory factors were detected.The changes in body weight,liver-to-body ratio and special circumstances during reproduction and development of mice in the two groups were observed.Results The offspring genotype of the target mice in the F2 generation was consistent with theoretical result of NLRP3flox/flox/AlbCre+/-.The mRNA and protein levels of NLRP3 in liver tissues of mice in the hepatocyte NLRP3 knockout group were significantly lower than those in the control group in the same litter(P<0.05).The mice in the hepatocyte NLRP3 knockout group was not affected in terms of growth,development and reproduction after the NLRP3 gene knockout.There were no statistically significant differences in the body weight,liver-to-body ratio,liver tissue morphology,serum liver transaminase or inflammatory factors between the hepatocyte NLRP3 knockout group and the control group in the same litter(P>0.05).Conclusion The Cre-LoxP gene knockout technology can be used to successfully construct a hepatocyte-specific NLRP3 gene knockout mouse model,providing an important technical support for the next step of studying the function of the NLRP3 gene in the liver at the animal level.

Objective: To analyze the effectiveness, safety and factors influencing the clinical prognosis of patients with hypopharyngeal carcinoma in T4b by nonsurgical treatments. Methods: The clinical data of 77 patients with T4b hypopharyngeal cancer treated in the College of Otolaryngology Head and Neck Surgery of the Chinese People's Liberation Army General Hospital from January 2010 to June 2021 were analyzed retrospectively. All were males, aged(57.0±8.0)years old. Patients were treated with induction chemotherapy plus concurrent chemoradiotherapy. Kaplan Meier survival analysis was used to compare the effects of different factors on prognosis. Adverse reactions during treatments and the causes of death were analyzed. Results: 98.7% of 77 patients with T4b hypopharyngeal cancer completed the chemotherapy plan and 94.8% completed the radiotherapy plan. The most common adverse reactions were grade 2 radiation oral mucositis (50/77, 64.9%) and grade 2 leukopenia (50/77, 64.9%). The incidence of grade 3 severe hoarseness was 7.8% (6/77), one patient (1.3%) underwent gastrostomy due to dysphagia, and pronunciation and swallowing function were effectively preserved in other patients. The overall survival rate was 71.9% at 1 year, 45.6% at 3 years and 29.7% at 5 years. The location of tumor, the presence of liquefaction necrosis in tumor, the use of molecular targeted drugs and the approach of radiotherapy were independent factors,each of which that affected the prognosis of T4b patients with advanced hypopharyngeal cancer [HR (95%CI) were 1.867(1.085-3.213), 3.018 (1.437-6.335), 0.372 (0.181-0.764) and 2.158 (1.015-4.588), respectively, P<0.05]. The two leading causes of death with high incidence were disease recurrence (12/32, 37.5%) and cervical large vessel rupture and hemorrhage (11/32, 34.4%). Conclusions: Non-surgical comprehensive treatment offers a high laryngeal preservation rate in patients with T4b hypopharyngeal cancer. The location of tumor, the liquefaction necrosis within tumor, the use of molecular targeted drugs, and the approach of radiotherapy are independent prognostic factors.
Aged ; Carcinoma, Squamous Cell ; Chemoradiotherapy ; Humans ; Hypopharyngeal Neoplasms/drug therapy* ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Abstract: Due to the continued emergence of multiple variants of SARS-CoV-2, the ongoing pandemic has resulted in severe mortality over the past two years. After the Alpha, Beta, Gamma and Delta variants, the most recent new variant of concern (VOC) strain to emerge is Omicron (B.1.1.529), which evolved as a result of the accumulation of a large number of mutations. The Omicron variant, which has a much higher transmission rate than the Delta variant, soon replaced the Delta variant and others, is now the dominant variant worldwide. The emergence of Omicron poses new challenges for the prevention and control of COVID-19 and has raised a number of concerns worldwide. Recently, cases of Omicron infection have been reported in several parts of China, and therefore this paper provides a comprehensive analysis and summary of the epidemiology and immune escape mechanisms of the Omicron variant. We also suggest some therapeutic strategies against the Omicron variant, including rapid diagnosis, genome analysis of emerging variants, ramping up of vaccination drives and receiving booster doses, updating the available vaccines, designing of multivalent vaccines able to generate hybrid immunity, up-gradation of medical facilities and strict implementation of adequate prevention and control measures need to be given high priority to handle the on-going COVID-19 pandemic successfully.

Objective:Explore the hydrogen-nuclear magnetic resonance ( 1H-NMR) application potential of metabolomics in early screening of vitamin A deficiency(VAD). Methods:From January to June 2019, 40 cases of left-behind children with confirmed VAD <1 year old were enrolled in Zunyi area as VAD group, and 40 healthy children were selected as healthy control group.Urine from the 2 groups of children was collected and 1H-NMR spectrum was measured.By combining metabonomics technology and multivariate statistical analysis, the 2 groups of metabolites were compared, and the differential metabolites of children with VAD were screened out. Results:Compared with VAD group [(0.56±0.10)μmol/L], the difference in the serum retinol concentration of healthy control group [(1.35±0.18)μmol/L] was statistically significant ( t=23.93, P<0.001). Compared with healthy group, VAD group showed higher level of methylmalonate, 2-ketoisovalerat, N-acetyl glycoprotein signal, dimethylglycine and betaine, with statistically significant differences (| r|>0.602, all P<0.05). The level of Isovalerylglycine, 2-hydroxyisobutyrate, creatine, phosphate choline/glycerophosphocholine, 4-PY and Hippurate were significantly decreased, with statistically significant differences (| r|>0.602, all P<0.05). Conclusions:Compared with healthy children, children with VAD have differences in urine Metabolites.Differential metabolites are closely correlated with intestinal microecological ba-lance, digestive system diseases, respiratory system diseases, immune-related diseases, energy metabolism, growth and development.Urine 1H-NMR metabolomics analysis has potential application value in the early screening of VAD.

Objective To evaluate the clinical efficacy of laparoscopic hepatectomy and open hepatectomy for the treatment of hepatocellular carcinoma.Methods Two hundred and sixteen patients with hepatocellular carcinoma who were admitted to the Chongqing Three Gorges Central Hospital from October 2014 to December 2017 were enrolled in this study,52 patients undergoing laparoscopic hepatectomy (laparoscopic group) and 164 patients undergoing open hepatectomy,52 patients (open group) were matched by propensity score matching method to reduce confounding bias.The operation time,tumor sizes,resection margin,time of hepatic inflow occlusion,volume of blood loss,blood transfusion,postoperative complication rate,drainage tube removal time,postoperative anal exhaust time,duration of postoperative hospital stay and survival rates were compared.T test was used for the measurement data in line with the normal distribution,and the results were represented by (Mean ± SD) tandard deviation;Wilcoxon test was used for the measurement data in the skewed distribution,and M(range) was used for represention,and the chi-square test or Fisher's exact probability test was used for the counting data.Kaplan-meier method was used to describe the survival curve,and log-rank test was used to compare the survival rate.Cox regression model was used to evaluate the risk factors affecting the survival of patients with hepatocellular carcinoma.Results All patients were recovered with no perioperative death.The operation time,tumor sizes,resection margin,time of hepatic inflow occlusion were (226.1 ±67.5) min,(49.2 ± 15.5) mm,(14.3 ± 4.9) mm,(34.1±18.1) min and (227.0 ±55.7) min,(49.4 ±16.6) mm,(14.6 ±4.0) mm,(38.2 ± 16.1) min,with no significant differences between the two groups(P ＞ 0.05).The volume of blood loss,blood transfusion,postoperative complication rate,drainage tube removal time,postoperative anal exhaust time,duration of postoperative hospital stay were 325(50-1 200) ml,11.5％,15.4％,(4.8 ±0.9) d,3(2-5) d,(11.9± 3.5) d and 375(100-2 000) ml,19.2％,23.1％,(5.9 ±0.8) d,4(3-5) d,(14.9 ±6.2) d,with significant differences between the 2 groups(P ＜0.05).1-and 3-year overall survival rates were 93.4％,83.7％ and 1-and 3-year disease-free survival rates were 87.7％,71.2％ in laparoscopic group;1-and 3-year overall survival rates were 94.1％,82.5％ and 1-and 3-year disease-free survival rates were 88.3％,70.8％ in open group,with no significant differences between the 2 groups(P ＞0.05).Cox regression analysis revealed that ICG R15,tumor size and tumor differentiation were independent risk factors of survival (hazard ratio ＞ 1,P ＜ 0.005).Conclusions Application of Laparoscopic hepatectomy for the treatment of selective hepatocellular carcinoma patients has better short-term efficacy than open hepatectomy and long-term efficacy is comparably.The propensity score matching can reduce confounding bias and have value for clinical research.

Objective: To explore the characteristic changes in urinary metabolites in left-behind children with vitamin D deficiency under 1 year old in Zunyi area by metabolomic nuclear magnetic resonance (NMR) in order to provide new biomarkers for early diagnosis of vitamin D deficiency. Methods: From January to August 2018, blood tests and urine collection were carried out on the left-behind children under 1 year old in Fenggang county, Bozhou district and Zheng′an county under Zunyi city by stratified sampling.Forty children diagnosed as a vitamin D deficiency were selected as a vitamin D deficiency group, and 40 children with normal urine test were selected as a healthy control group.For urine sampling, SIMCA-P+ software was applied to analyze the integral value of hydrogen spectrogram by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) was used to distinguish the difference in urine metabolites between two groups of the left-behind children.Orthogonal partial least squares discriminant analysis (OPLS-DA) was used to screen different metabolites. Results: The serum level of 25-hydroxy vitamin D[25-(OH)D][(32.0±3.6) nmol/L ] in the healthy control group was higher than that in the vitamin D deficiency group[(15.8±2.3) nmol/L], and the difference was statistically significant (P<0.05). PCA and PLS-DA analysis showed significant differences in urine metabolites between the healthy control group and the vitamin D deficiency group (P<0.05). OPLS-DA indicated R²X=0.365, Q²=0.978, which further verified the difference of metabolites.Compared with the healthy control group, the urine of methyl malonic acid, 3-hydroxy butyrate, N-acetyl glycoprotein signal, glutamic acid, dimethyl glycine, 2-ketone glutaric acid, taurine, fumaric acid salt level increased significantly in the vitamin D deficiency group, and the differences were statistically significant (|r|>0.602, all P<0.05, df=39). However, the levels of ethyl malonic acid, creatine, choline, glycerophosphalocholine and equine were significantly decreased, and the differences were statistically significant (|r|>0.602, all P<0.05, df=39). Conclusions The left-behind children under 1 year old with vitamin D deficiency in Zunyi region are mainly characterized by disorder in energy metabolism, lipid metabolism, amino acid metabolism and intestinal microbial meta-bolism disorders, and their differential metabolites have potential application value in early diagnosis and pathogenesis of vitamin D deficiency.

ObjectiveTo evaluate the integrated performance of age, serum PSA, and transrectal ultrasound images in the prediction of prostate cancer using a Tree-Augmented NaÏve (TAN) Bayesian network model.

METHODSWe collected such data as age, serum PSA, transrectal ultrasound findings, and pathological diagnoses from 941 male patients who underwent prostate biopsy from January 2008 to September 2011. Using a TAN Bayesian network model, we analyzed the data for predicting prostate cancer, and compared them with the gold standards of pathological diagnosis.

RESULTSThe accuracy, sensitivity, specificity, positive prediction rate, and negative prediction rate of the TAN Bayesian network model were 85.11%, 88.37%, 83.67%, 70.37%, and 94.25%, respectively.

CONCLUSIONSBased on age, serum PSA, and transrectal ultrasound images, the TAN Bayesian network model has a high value for the prediction of prostate cancer, and can help improve the clinical screening and diagnosis of the disease.

Bayes Theorem ; Biopsy ; Humans ; Male ; Predictive Value of Tests ; Prostate ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; diagnosis ; Sensitivity and Specificity

This study was to establish a model to explore anti- RSV effect of different administration method of Chinese medicine realgar on respiratory syncytial virus type A (RSV-A) replication in Hep-2 cells. Using high-energy ball milling with distilled water to prepare realgar nanoparticles,the concentration of nanometer realgar was tested by molybdenum blue staining method and the size of realgar nanoparticles was tested on Nano Series. Cell culture with ribavirin as a positive control was applied to observe the effect of anti-respiratory syncytial virus type A replication through prevention, treatment or direct inactivation of three different drug administration methods. Realgar nano-particles was found to be a potential inhibitor of RSV-A in a concentration-dependent manner with the median toxic concentration(TC50) of 0.649 microg/mL in Hep-2 cell culture. The median inhibition concentration (IC50) was 0.20 microg/mL when drug was added before virus infection. The IC50 was 0.13 microg/mL when drug was added after virus infection,and it was 0.16 microg/mL when the drug was mixed with virus and added. The therapeutic index (TI) was 3.18, 4.99 and 4.11, respectively. The results showed realgar nanoparticles could inhibit the replication of the RSV and inactivate the RSV in vitro.
Arsenicals ; pharmacology ; Nanoparticles ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Sulfides ; pharmacology ; Virus Replication ; drug effects

Objective To evaluate the efficacy and safety of ligation-assisted endoscopic dissection (ED-L) technique for the removal of gastric tumors originating from muscularis propria.Methods A total of 33 patients with gastric tumors originating from muscularis propria less than 10 mm were treated with ED-L procedures.The tumor was ligated by elastic bands.Endoscopic dissection was performed until the tumor was partially or completely dissected from muscularis propria by using Hook knife and/or IT-knife.The wound was closed with metallic clips and medical adhesive.The patients were followed up 1 week,1 month,3 months,6 months and 12 months thereafter with endoscopy,respectively.Results Of the 33 gastric tumors,there were 25 partial dissections and 8 complete dissections.All of the tumors sloughed completely.Pathological diagnoses of all the patients were acquired.No complications like perforation occurred except for one self-limiting and non-life-threatening hemorrhage.There was no recurrent case during the 3-18 months of follow-up period.Conclusion ED-L is a safe,effective and relatively simple technique for excision of small gastrointestinal tumors originating from muscularis propria,providing a histopathological diagnosis as well.