Next generation sequencing(NGS)has made great progress, applied from science technology to clinical diagnosis in the past several years.With a simple, fast, high-throughput, and high-resolution feature, it has been believed to be one of the most potential technology in clinical tests, especially in individual tumor diagnosis and treatment, noninvasive prenatal screening, genetic disorders screening field.This paper will briefly review the application of next generation sequencing in the field of infection disease.

Objective To introduce a novel technique of intracolonic shunt procedure used in the anus - preserving operation for acute intestinal obstruction resulted from cancer at low and middle portions of rectum and assess the clinical significance. Methods In total, 81 patients with acute obstruction of low and middle portion of rectum caused by cancer were randomly ( random number) divided into control group and study group. In control group, 42 patients were operated with preventive transverse colonostomy or terminal ileum stoma after low proximal resection of rectum involved in cancer, while 39 patients were operated with intracolonic shunt procedure by using a biodegradable anastomosis ring and a condom placed 5 cm above anastomosis for protection in study group. Results There were no significant differences in sex, age, tumor site, tumor size and the distance from anstomosis to anal-edge between two groups. In both groups, the bowel movement resumed in 2 ~ 5 days after operation (P > 0.05). In study group, the rate of anastomosis leakage was 7.7% (3/39), and leakages were treated with drainage for 7.1 days in average to be healed, and the biodegradable anastomosis ring detached and were discharged in 14 -23 days (17 days in average), and there were no complications of drainage happened. The anastomotic stenosis occurred in three patients (7. 7% ) within 6 months after operation. In control group, 11.9% patients (5/42) had anastomosis leakage and they treated with drainage for 18.2 days in average to get the leakage healed, and 35. 7% patients (15/42) had stoma complications, and anastomotic stenosis happened in 28.6% patients (12/42) within 6 months after operation, and 7. 1% patients need another operation because of severe anastomosis stenosis. There were no significant differences in rate of anastomosis leakage between tow groups ( P > 0. 05), but there were significant differences in drainage days after anstomosis leakage happened and 6 - months anastomosis stenosis between two groups (P<0.05). Conclusions In the anus -preserving operation for acute intestinal obstruction at low and middle portions of rectum caused by cancer , the intracolonic shunt procedure is convenient and safty, and reduces the hazard incurred by anastomosis leakage and anastomosis stenosis compared with classic stoma operation.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.

Objective:The purpose of this study is to use the next-generation sequencing (NGS) technology platform to detect the methylation rate of phosphatase and tensin homolog deleted on chromosome ten ( PTEN) promoter region in hepatocellular carcinoma (HCC) tissue samples, and to analyze the clinical significance of its correlation with the prognosis of patients receiving sorafenib treatment. Methods:The 52 pairs of tumor tissue and para-cancerous tissue samples from HCC patients treated with sorafenib alone, which were collected and preserved in the Liver Tumor Diagnosis and Research Center of the former 302 Hospital of the People′s Liberation Army by the National Natural Science Foundation of China Youth Project with the project batch number 81702986 in 2018, were extracted total DNA from the samples. Then the DNA samples were treated with bisulfite and specific primers were designed to amplify the PTEN promoter region. Finally, the amplified products were analyzed by second-generation sequencing. In the analysis of clinical significance of PTEN methylation, log-rank statistical analysis was used to calculate whether there was a statistical difference in survival between the patient groups. Results:The methylation rate of PTEN promoter region in tumor tissues (29.17%±9.58%) was significantly higher than that in paracancer tissues (4.17%±2.86%)( t=19.970, P<0.05). At the same time, in HCC tissues, the methylation rate of the PTEN promoter region is negatively correlated with its expression ( F=47.270, P<0.000 1; Y=-1 800× X+38.03), and the PTEN methylation rate is negatively correlated with the prognosis of patients receiving the molecularly targeted drug Sorafenib (χ2=4.313, P<0.05). Conclusion:This study successfully established a new method for detecting methylation in the promoter region of PTEN, and the methylation rate of PTEN can be used as one of the targets of HCC diagnosis and targeted therapy.

Autoimmune disease is a kind of immune dysfunction disease caused by gene and environmental factors. At present, there are some difficulties in early diagnosis, prognosis evaluation and curative effect evaluation. More and more autoantibodies have been found and confirmed to be closely related to the occurrence and development of autoimmune diseases with the deepening of related research. Accurate autoantibody detection is helpful to improve the diagnostic efficiency, therapeutic effect and quality of life of patients with autoimmune diseases.

Objective:This study will analyze the clinical characteristics and risk factors that may be related to the 30-day mortality of patients infected with CRAB in intensive care unit (ICU), and explore the resistance of CRAB and its influence on mortality.Methods:From December 2012 to February 2021, 173 ICU patients with CRAB infection in the Fifth Medical Center of PLA General Hospital were selected as the research objects, and the relevant data were collected for retrospective analysis. There were 119 cases (68.8%) in survival group and 54 cases (31.2%) in the non-survival group. Patients with CRAB infection were (52.9±13.5) years old, including 140 males (80.9%) and 33 females (19.1%).The first detected CRAB was collected, and antibiotic sensitivity test was conducted after the strain was resuscitated to analyze the antibiotic resistance. Univariate and multivariate Cox models were used to analyze independent risk factors associated with 30-day mortality in patients with CRAB infection.Results:Univariate and multivariate Cox analysis showed that acute physiology and chronic health evaluation scoring system Ⅱ(APACHE Ⅱ)(HR=1.058, 95% CI:1.012-1.106, P=0.013) and septic shock (HR=6.240, 95% CI:2.227-17.483, P<0.001) were independent risk factors related to 30-day mortality in ICU patients with CRAB. Treatment with β-lactamase inhibitor (HR=0.496, 95% CI: 0.275-0.893, P<0.019) can reduce the 30-day mortality of patients with CRAB infection in ICU. The resistance rate of CRAB to cephalosporins, carbapenems, aminoglycosides and quinolones were more than 80%. The survival rate of patients infected by aminoglycoside resistant CRAB is low(χ2=4.012, P<0.05). Conclusion:The APACHE Ⅱ score, septic shock and use of β-lactamase inhibitors were independent factors associated with the 30-day mortality in ICU patients with CRAB infection.

Objective:This study will analyze the clinical characteristics and risk factors that may be related to the 30-day mortality of patients infected with CRAB in intensive care unit (ICU), and explore the resistance of CRAB and its influence on mortality.Methods:From December 2012 to February 2021, 173 ICU patients with CRAB infection in the Fifth Medical Center of PLA General Hospital were selected as the research objects, and the relevant data were collected for retrospective analysis. There were 119 cases (68.8%) in survival group and 54 cases (31.2%) in the non-survival group. Patients with CRAB infection were (52.9±13.5) years old, including 140 males (80.9%) and 33 females (19.1%).The first detected CRAB was collected, and antibiotic sensitivity test was conducted after the strain was resuscitated to analyze the antibiotic resistance. Univariate and multivariate Cox models were used to analyze independent risk factors associated with 30-day mortality in patients with CRAB infection.Results:Univariate and multivariate Cox analysis showed that acute physiology and chronic health evaluation scoring system Ⅱ(APACHE Ⅱ)(HR=1.058, 95% CI:1.012-1.106, P=0.013) and septic shock (HR=6.240, 95% CI:2.227-17.483, P<0.001) were independent risk factors related to 30-day mortality in ICU patients with CRAB. Treatment with β-lactamase inhibitor (HR=0.496, 95% CI: 0.275-0.893, P<0.019) can reduce the 30-day mortality of patients with CRAB infection in ICU. The resistance rate of CRAB to cephalosporins, carbapenems, aminoglycosides and quinolones were more than 80%. The survival rate of patients infected by aminoglycoside resistant CRAB is low(χ2=4.012, P<0.05). Conclusion:The APACHE Ⅱ score, septic shock and use of β-lactamase inhibitors were independent factors associated with the 30-day mortality in ICU patients with CRAB infection.

Objective:To evaluate the positive rate of nucleic acid detections in throat swab and stool specimens of patients, thereby to provide a reference to the clinical laboratory diagnosis.Methods:Forty-one confirmed patients were enrolled in this study, their throat swab and stool specimens were collected for detections. All the samples were determined by a fluorescence quantitative PCR method . The positive rate and concordance were evaluated for these two types of samples.Results:Among those 41 patients, 9 of them had positive result in throat swab samples, accounted for 30%. Five patients had positive result in stool samples, accounted for 12.2%. Only two patients had concordant positive result . Kappa tests showed that the concordance was poor, kappa index was 0.153. Conclusions:The nucleic acid detection in stool specimen is a complemental method for respiratory tract samples and therefore may provide. additional information for infectiousness of patients.

Objective To compare the changes of the medical animal resources(MAR)in Sichuan based on the data of the 3rd Chinese Materia Medica Resource Inventory(CMMRI,1983-1986)and the 4th CMMRI(2020-2022).Methods After field investigation and identification of the photos of the animals,the data of the MAR in Sichuan found in the 4th CMMRI were analyzed and compared with the data of 3rd CMMRI.Results 745 species of MAR were found in Sichuan during the 4th CMMRI,including 212 families and 468 genera.Compared with the 108 species found in 3rd CMMRI,the number of MAR in Sichuan had greatly increased,The Aves was found to have 243 species of MAR,which is the most plenty one among the 7 classes.There were 14 families which have more than 10 species of MAR.The family,Cyprinidae had 48 species of MAR.There were 33 common Chinese medicinal herbs and 3 genuine medicinal materials including Cordyceps sinensis,musk and Venenum bufonis were found in this investigation.The new distributions of Liangshan Cordyceps and Cordyceps gunnii were found in the investigation,and the new resources of Atypus heterothecus was found in Mountain Emei.There were 140 species of key protection of wild medicinal animals,including 44 species of animals under first-class protection and 96 species of animals under second-class protection.There were 230 species of animals which had important ecological,scientific,and social values,too.Conclusion Sichuan was plenty of medical animal resources and the protection of the forest musk deer,the Cordyceps sinensis and the pangolin was need to be strengthened.