Objective: Investigate osteogenic differentiation of canine periodontal ligament stem cells ( cPDLSCs) via over-expression ephrinB2 in cPDLSCs． Methods : cPDLSCs were isolated from the premolars and molars of Beagle．After transfected with EfnB2-GFP-Bsd and GFP-Bsd empty Vector，cPDLSCs were induced to osteogenic differentiation．Western blot was used to invest the expression of ephrinB2 protein．The effect of osteogenic differentiation of EfnB2-cPDLSCs and Vector-cPDLSCs were analyzed by RT-PCR , CCK-8，Alizarin-red S staining and ALP. Results: There was no significant difference in cell proliferation between EfnB2-cPDLSCs and Vector-cPDLSCs．While EfnB2-cPDLSCs displayed an enhanced ALP activity and more prominent mineralized nodules compared with Vector-cPDLSCs．The odonto-/ osteogenic genes in EfnB2-cPDLSCs were also highly enhanced． Conclusion The results of our study indicated that ephrinB2 gene-transfected cPDLSCs showed enhanced osteogenic differentiation.

Objective:To screen the HIV standard strains with typical biological characteristics of HIV strains circulating in China through the isolation, culture, genotype and phenotype identification of HIV from the whole blood samples of HIV-infected persons, confirm genetic characteristics, traceability, and in line with the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023).Methods:Whole blood samples were collected from 48 HIV infected patients. Peripheral blood mononuclear cells (PBMCs) were isolated from the samples and co-cultured with PBMCs isolated from healthy persons′ whole blood samples to isolate and culture HIV from infected persons. We determined concentration of p24 antigen and the virus titer in the culture supernatant. The viral RNA was extracted from the successfully isolated strains, and the gag, pol genes and env C2V3 fragments of the viral genome were amplified and sequenced. The genotype, gene recombination and drug resistance sites were determined according to the viral gene sequences. Virus infection and replication were monitored by inoculating the virus culture supernatant into Ghost cells expressing CCR5 or CXCR4 to determine the viral tropism.The formation of syncytium was observed by inoculating the virus culture supernatant into MT-2 cells to determine whether was a syncytium-induced phenotype. Results:Fourteen strains with p24 antigen concentration > 1 ng/ml in culture supernatant were isolated and cultured from 48 fresh EDTA anticoagulated whole blood samples of HIV infected persons. Of the 14 strains, only one strain with a titer≥10 5 TCID 50/ml, 8 strains with titers ≥10 4 TCID 50/ml, and the other 5 strains with titers≥10 3 TCID 50/ml. Phylogenetic analysis showed that the genotypes of the strains were 9 strains of subtype B, 3 strains of CRF01_AE and 2 strains of CRF07_BC recombinant. Genotypic resistance analysis showed that 11 strains contained drug resistance sites. Ghost cells were used to verify the tropism of the strains, and it was found that 8 strains were CCR5 tropism, 6 strains were CXCR4 & CCR5 dual tropism. Only 2 of the 14 strains could induce MT-2 cytopathic effect, which was syncytium-inducing phenotype. Conclusions:Fourteen HIV strains with typical biological and genetic characteristics were isolated to screen the standard HIV strains. Among which, 1 strain was evaluated as a standard HIV strain that meets the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023). This study can also provide technical guidance for the screening of the HIV standard strains. Next step is to complete the application and reserve database construction according to the sharing mechanism of the HIV standard strains, to provide resources for the researches of HIV vaccines and drugs.

BACKGROUND: Perioperative treatment has become an increasingly important aspect of the management of patients with non-small cell lung cancer (NSCLC). Small-scale clinical studies performed in recent years have shown improvements in the major pathological remission rate after neoadjuvant therapy, suggesting that it will soon become an important part of NSCLC treatment. Nevertheless, neoadjuvant immunotherapy may be accompanied by serious adverse reactions that lead to delay or cancelation of surgery, additional illness, and even death, and have therefore attracted much attention. The purpose of the clinical recommendations is to form a diagnosis and treatment plan suitable for the current domestic medical situation for the immune-related adverse event (irAE). METHODS: This recommendation is composed of experts in thoracic surgery, oncologists, thoracic medicine and irAE related departments (gastroenterology, respirology, cardiology, infectious medicine, hematology, endocrinology, rheumatology, neurology, dermatology, emergency section) to jointly complete the formulation. Experts make full reference to the irAE guidelines, large-scale clinical research data published by thoracic surgery, and the clinical experience of domestic doctors and publicly published cases, and repeated discussions in multiple disciplines to form this recommendation for perioperative irAE. RESULTS: This clinical recommendation covers the whole process of prevention, evaluation, examination, treatment and monitoring related to irAE, so as to guide the clinical work comprehensively and effectively. CONCLUSIONS Perioperative irAE management is an important part of immune perioperative treatment of lung cancer. With the continuous development of immune perioperative treatment, more research is needed in the future to optimize the diagnosis and treatment of perioperative irAE.

Objective:To investigate the nutritional status of elderly inpatients in China, and to assess its relationship with clinical outcomes.Methods:A prospective, multicenter, parallel investigation was organized and conducted by the Chinese Medical Association's Group of Geriatric Nutrition Support.Patients aged ≥65 years from 30 major hospitals of 14 cities in China were evaluated by the Nutritional Risk Screening 2002(NRS2002)and the Mini-Nutritional Assessment-Short Form(MNA-SF), in order to understand the nutritional status and nutritional risk of elderly inpatients in China.The indicators of clinical outcomes were summarized, and the correlation between nutritional status and clinical outcomes was analyzed.Results:A total of 10 184 elderly patients who met the inclusion criteria were enrolled in this study, aged 65-112(74.81±7.01)years, with a body mass index(BMI)of 17.80-35.50(23.32±3.83)kg/m 2.Grip strength of the dominant hand was(16.95±18.42)kg, upper arm circumference was(25.68±3.70)cm, and calf circumference was(32.07±3.89)cm.BMI, grip strength, upper arm circumference and calf circumference decreased significantly with age( F=13.74, 97.47, 28.31 and 88.68, all P<0.001). NRS2002 was conducted on 10 182 patients.Of them, 10.14%(895/10 182)suffered malnutrition(BMI≤18.5 kg/m 2), and 46.42%(4 726/10 182)were at nutritional risk(NRS2002 score≥3). Nutrition deficiency and nutritional risk showed upward trends with age( F=43.41 and 177.05, both P<0.001). A total of 9 755 patients(95.79%, 9 755/10 182)completed the MNA-SF.Of them, 14.67%(1 431/9 755)had malnutrition, 35.04%(3 418/9 755)were at risk of malnutrition, and 50.29%(4 906/9 755)had normal nutritional status.The incidence of malnutrition and the risk of malnutrition significantly increased with age( F=172.79, 12.10 and 152.42, all P<0.05). Nutritional risk(NRS2002 score≥3)was related to age, BMI, mortality, infectious complications, length of hospital stay and total hospital cost(all P<0.05). Conclusions:The incidence of malnutrition and the risk of malnutrition are high in elderly inpatients in China.Nutritional risk is an influencing factor for adverse clinical outcomes.

Objective: To learn the population,virus status and viral types of hantavirus（HV）hosts in Tiantai County of Zhejiang Province from 2011 to 2018,and to provide evidence for hemorrhagic fever with renal syndrome（HFRS）control. Methods: Rodents in Tiantai County were captured by night trapping method. After the species and age of rodents were identified,the composition of rodent species,dominant species and density of rodents were analyzed. The lungs and blood of rodents were sampled to detect the antigen and antibody of HV by immunofluorescence method. The HV antigen-positive lung samples were detected by RT-PCR with specific primers of HV S fragment,then HV was isolated and identified by inoculating Vero-E6 cells. Results: The average rodent density in Tiantai County from 2011 to 2018 was 4.44%. The rodent density in the field and residential areas were 4.94% and 2.23%,respectively. Ten species of rodents were identified,with Apodemus agrarius dominant in the field and Rattus norvegicus in the residential areas. Sixty-seven lung samples were HV antigen positive（4.13%）,one from Rattus norvegicus and the other sixty-six from Apodemus agrarius. Seventy-nine blood samples were HV antibody positive（4.86%）,all from Apodemus agrarius. Thirty-four HV antigen-positive lung samples were positive（50.75%）after RT-PCR amplification. Twenty-two strains of virus were isolated and all of them were from Apodemus agrarius,including twenty-one strains of Hantaan type（HTN）and one strain of Seoul type（SEO）. Conclusion In Tiantai County,Apodemus agrarius is the main source of HFRS infection;the main epidemic type of HV is HTN and SEO is first found in Apodemus agrarius.

Objective: By investigating the genotype and evolutionary variation of hantavirus (HV) in Tiantai county, a national surveillance site for hemorrhagic fever with renal syndrome (HFRS) was set in Zhejiang province, from 2011 to 2018, to reveal the molecular epidemiological characteristics of hantavirus (HV) in Tiantai. Methods: Total RNA was extracted from ultrasound treated HV antigen- positive rat lung samples in Tiantai from 2011 to 2018. After cDNA was prepared, nested PCR was used to amplify partial sequence of M fragments by using specific primers of HV. The sequences of HV in Tiantai from 2011 to 2018 were compared with other known HV sequences in order to identify the genotype and analyze the evolution and variation of the virus. Results: In 67 HV antigen-positive lung specimens, 31 were positive in nested PCR amplification with type-specific primers, including 30 Hantaan virus (HTNV) positive samples, 1 Seoul virus (SEOV) positive sample, and all the 31 samples were from Apodemus agrarius. The phylogenetic tree based on partial M segment was divided into monophyletic group, 30 strains were distributed in HTNV group and 1 was in SEOV group. The HTNV strain Tiantai T2018-130 was independently in one branch, sharing 84.8%-87.9% homology with other strains both at home and abroad, including 29 strains in HTNV group in Tiantai. The other 29 HTNV strains in Tiantai showed closer relationship. The SEOV strain T2016-31 from Apodemus agrarius showed closer relationship with previous strains of SEOV, Tiantai ZT71, ZT10 and Z37 strains of Wenzhou, Zhejiang province. Conclusions HTNV, the main genotype of HV in Tiantai of Zhejiang province, showed obvious geographic clustering, but the strain T2018-130 was distinct from the others in Tiantai. Meanwhile, by sequence analysis, we confirmed that The SEOV strain T2016-31 existed in in Apodemus agrarius, indicating there was a phenomenon of "spillover" between virus and host in SEOV evolution.

OBJECTIVE: To study the effect of CYP2D610 (c.100 C>T) on plasma trough concentrations of metoprolol and its metabolite α-hydroxy metoprolol, blood pressure and heart rate in patients with coronary artery disease. METHODS: The patients with coronary artery disease taking metoprolol tablets (=128) and those taking metoprolol sustained-release tablets (=126) were genotyped for CYP2D610 using Taqman real-time quantitative PCR. The trough concentrations of metoprolol and α-hydroxy metoprolol were determined with UPLC-MS/MS, and the dose-normalized concentrations (C/D) were compared among the patients with different CYP2D610 genotypes in both groups. Resting blood pressure and heart rate were recorded in all the patients when the concentration of metoprolol reached the steady state and were compared among the patients with different genotypes. RESULTS: In patients taking metoprolol sustained-release tablets, the plasma trough concentration of α-hydroxy metoprolol was significantly associated with the systolic blood pressure (=0.0204). The CYP2D610 poor metabolizers showed a significant association with the C/D of metoprolol and α-hydroxy metoprolol ( < 0.01) in patients receiving metoprolol in both formulations, and in both groups, the C/D of metoprolol was significantly higher in the patients with a TT genotype than in those with a CC or CT genotype ( < 0.01); compared with those with the CT genotype, the patients with the TT genotype had a significantly lower C/D of α-hydroxy metoprolol ( < 0.01). In patients taking metoprolol sustained-release tablets, those with the CT (=0.0281) and TT (=0.0196) genotypes had lower diastolic blood pressure than patients with the CC genotypes, but the systolic blood pressure or heart rate did not differ significantly among them. CONCLUSIONS CYP2D610T allele mutation can reduce the metabolism of metoprolol, increase the C/D of metoprolol and decrease the C/D of α-metoprolol and diastolic blood pressure in patients with coronary artery disease, but CYP2D610 variation does not significantly affect systolic blood pressure or heart rate in the patients when the concentration of metoprolol reaches a steady state.
Adrenergic beta-Antagonists ; Chromatography, Liquid ; Coronary Artery Disease ; Cytochrome P-450 CYP2D6 ; Genotype ; Humans ; Metoprolol ; Tandem Mass Spectrometry

As a new type of anti-cancer drugs, immune checkpoint inhibitors (ICIs) have shown remarkable therapeutic effects in many malignant tumors. By virtue of their targets and mechanisms of action, ICIs can cause autoimmune and inflammatory effects, termed immune-related adverse events (irAEs). Unlike traditional therapies, irAEs are latent and unstable. Severe adverse reactions will force patients to discontinue treatment and even affect their survival. Therefore, with the wide application of ICIs in clinical practice, clinicians need to fully understand the possible adverse reactions of such drugs and appropriate treatment strategies, in order to improve the survival rate and treatment effect of patients receiving ICIs. In this article, we review the incidence, clinical features, diagnosis and treatment of immune-related endocrine events that may occur with administration of ICIs.

Objective To investigate the application value of mini-nutritional assessment short-form(MNA-SF) in screening and evaluating nutritional status in elderly hospitalized patients with malignant tumor.Methods 1472 elderly hospitalized patients(≥65 years old)with malignant tumor were enrolled and prospectively studied for evaluating the nutritional status by nutritional risk screening 2002 (NRS 2002)assessment.Indicators of clinical outcomes were summarized.The consistencies of MNA-SF with body mass index(BMI),grip and results of NRS 2002 assessment,and the relationship between the different nutritional status and clinical outcomes were analyzed.Results The patients meeting the inclusion/exclusion criteria were aged(72.1±5.9)years,with BMI of(22.3 ±3.7)kg/m2 and grip of(21.7± 19.0)kg.Among 16.7％ of the malnourished patients with malignant tumors,pancreatic cancer (30.9 ％),bile duct cancer (24.1％) and esophageal cancer (21.2 ％) occupied the top three incidence of malnutrition,with the lowest (5.4％)incidence in breast cancer.Among 59.2％ of patients being malnourished (16.7％)or at risk(42.5％)of malnutrition,the highest incidence was in bile duct cancer(82.8％),and lowest one was in breast cancer(28.6 ％).MNA-SF-discriminated malnutrition(0～ 7 points)showed a great agreement with malnutrition evaluation by BMI(＜18.5),but had a poor agreement with grip in screening malnutrition.MNA-SF-discriminated nutrition problem(malnourishment plus at risk of malnutrition)showed a great agreement with NRS 2002-discovered nutrition problem (score≥3).Under-nourished patients had a higher infection complication(9.29％ vs.5.14％,P =0.006)and longer hospital stays(15.4 d vs.12.8 d,P＜0.01)as compared with patients with normal nutrition status.Conclusions The prevalence and risk factors for malnutrition are higher in elderly patients with malignant tumor.Poor nutritional status is correlated with poor clinical outcomes.MNA-SF can be used as a tool for evaluating the nutritional status of elderly patients with malignant tumor.

Objective To compare the long term effectiveness of tonsillotomy(TT)vs. tonsillectomy (TE)by hypercator in treatment of obstructive sleep apnea hypopnea syndrome(OSAHS)in children. Methods 119 children with tonsil hypertrophy were diagnosed with OSAHS and included in the study. Among them,57 children underwent TT by hypercator and 62 TE only after preoperative laboratory polysomnography(PSG). All participants were interviewed 5 years after operation to assess the satisfaction of their caregivers,investigate the frequency of respiratory infections,the rate of tonsil regrowth and evaluate their quality of life by Obstructive Sleep Apnea-18(OSA-18). Results Some of them received postoperative PSG. All parents reported alleviation of breathing obstruction,but the postoperative satisfaction in TT group was higher than TE group mainly because of the shorter time return to normal diet. The rate of the respiratory tract infections was not increased in all cases after surgery. Four cases of children were observed with tonsillar regrowth,but no surgery was needed for further treatment. The rate of the dry throat symptoms in TT group was lower than TE group. Conclusions Tonsillotomy by hypercator can effectively eliminate the obstruction symptoms in children with OSAHS,superior to TE for its shorter recovery period and smaller oral mucosal injury and lower rate of tonsils regrowth and it is worth to clinical promotion.