Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

ObjectiveTo evaluate the efficacy of three screening questionnaires for COPD in the community residents of Songjiang District， Shanghai， and to provide a basis for selecting COPD screening questionnaire and process that are more suitable. MethodsCommunity residents aged 40 years or over were randomly selected from the Shanghai Suburban Adult Cohort and Biobank for the study with screening questionnaires and spirometry. Questionnaires included the COPD screening questionnaire （COPD-SQ）， the COPD population screener （COPD-PS） and the revised COPD diagnostic questionnaire （revised-CDQ）. Evaluation of the efficacy of these questionnaires was based on the area under the receiver operating characteristic curve （AUC） of the subjects. DeLong test was used to compare the accuracy of different questionnaires； Z test was used to compare the accuracy of different cut-off values for the same questionnaire. ResultsAmong 3 184 community residents， a total of 259 （8.1%） COPD patients were screened by spirometry. AUC values of these 3 screening questionnaires were >0.7 indicating that they were reliable COPD screening tools. The sensitivity and specificity of the questionnaires at the recommended cut-off values were COPD-SQ （63.7% and 72.2%）， COPD-PS （12.0% and 96.1%）， and revised CDQ （78.8% and 52.7%）， with the COPD-SQ having the highest screening accuracy （AUC=0.754）. The optimal and recommended cut-off values for the three questionnaires differed in this population， but the difference in accuracy was statistically significant only for COPD-PS. The optimal cut-off values for the three questionnaires differed between male and female， and the sensitivity and accuracy of COPD-SQ and COPD-PS improved when lower cut-off values were used for women. The AUC was greater when two questionnaires were utilized simultaneously for screening， but the differences were not statistically significant. ConclusionThe COPD-SQ is recommended for primary COPD screening； a lower cut-off value for women should be considered. The COPD screening questionnaire needs to be further improved for the early diagnosis and treatment of COPD patients.

Objective:To analyze the clinicopathological features of solitary papillary thyroid isthmus carcinoma (SPTIC) and the therapeutic effect and prognosis of different surgical methods.Methods:A total of 161 patients with SPTIC who underwent surgical treatment in the Department of Thyroid Surgery of the First Hospital of Jilin University from Dec. 2012 to Oct. 2021 were selected. Gender, age, body mass index and pathology of the patients were collected and the clinicopathological characteristics of SPTIC were analyzed. They were divided into three groups according to different surgical methods: group A underwent isthmic excision, group B underwent extended isthmic excision, and group C underwent total thyroidectomy. There were 47 patients in group A (8 males and 39 females with an average age of 42.6±9.1 years), 50 patients in group B (11 males and 39 females with an average age of 45.3±11.3 years), and 64 patients in group C (10 males and 54 females with an average age of 46.9±11.4 years). The clinicopathological features and therapeutic effect of the three groups were compared by ANOVA, multiple local rank sum test, χ2 test or Fisher's exact probability method, and the recurrence rate and recurrence free survival (RFS) of the three groups were compared after a follow-up of 6 to 126 months. Results:Among the 161 patients with SPTIC, 132 (82.0%) were female, 130 (80.7%) were younger than 55 years old, BMI (25.1±3.6) kg/m 2, 124 (77.0%) were combined with capsule invasion. There were 53 cases (32.9%) with central lymph node metastasis (CLNM). Subgroup analysis showed that the proportion of males in patients with tumor diameter > 1 cm, the proportion of patients with BMI≥26.0 kg/m 2, the rate of capsular invasion, the rate of extrandular invasion and the rate of CLNM were higher (30.0% vs. 14.0%, P=0.023; 60.0% vs. 33.9%, P=0.004; 97.5% vs. 70.2%, P< 0.001; 42.5% vs. 9.9%, P<0.001; 50.0% vs. 27.3%, P=0.008) ; The CLNM rate was higher in male patients with BMI≥26.0 kg/m 2 and tumor diameter > 1 cm (28.3% vs. 13.0%, P=0.017; 52.8% vs. 34.3%, P=0.024; 37.7% vs. 18.5%, P=0.008). Compared with groups A and B, group C had longer hospitalization days, higher hospitalization costs, longer operation time, more postoperative drainage flow and higher incidence of postoperative hypoparathyroidism, and the differences were statistically significant (all P< 0.05). Groups A and B were similar in all aspects, with no statistical significance ( P> 0.05 for all). During the follow-up, 3 patients in both group A and B relapsed, while no patients in group C relapsed, and there were no statistically significant differences in the recurrence rate ( P=0.059) or RFS ( P=0.082) among the three groups. Conclusions:The majority of SPTIC were microcarcinomas, but the incidence of tumor combined with capsule invasion was higher, and the tumor size of more than 1 cm was more invasive (capsule invasion rate and extrathyroid invasion rate were higher). SPTIC should be treated as conservatively as possible (isthmic excision or enlarged isthmic excision) .

Objective:To explore the safety and efficacy of laparoscopic right posterior sectionectomy (LRPS) in the left side lying at 60° run-hug position.Methods:The clinical data of 12 cases underwent LRPS at the Department of Biliary and Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University from August 2022 to June 2023 were analyzed. There were 9 males and 3 females, ranging in age from 26 to 76 years old. To evaluate the safety and effectiveness of the operation, operation time, intraoperative blood loss, the number and time of intraoperative hilar occlusion, postoperative drainage time, postoperative hospitalization time and postoperative complications were summarized.Results:Twelve operations were successfully completed under laparoscopy, and no patient was converted to laparotomy due to the difficulty in controlling intraoperative bleeding. 12 patients included 1 case of intrahepatic bile duct stones, 5 cases of primary liver cancer, 2 cases of hepatic hemangioma, 2 cases of focal nodular hyperplasia of the liver, 1 case of metastatic liver cancer, and 1 case of liver abscess. Operation time was (246.2±90.4) min, and the intraoperative blood loss was 125(22, 275) ml. The number of intraoperative hilar occlusion was 2.00(0.25, 2.75), and intraoperative hilar occlusion time was (27±22) min. Postoperative drainage time was (5.0±2.6) d, and the length of postoperative hospital stay was 6.0(4.3, 8.0) d. Intraoperative blood transfusion occurred in 1 patient, and postoperative abdominal effusion occurred in 1 patient. There was no postoperative death. All patients were recovered and discharged successfully.Conclusion:LRPS is safe and effective for the lesions in the right posterior hepatic region with the left side lying at 60° run-hug position.

Objective:To investigate the characteristics and surgical management of mesh infection after tension-free repair of inguinal hernia.Methods:The clinical and follow-up data of 87 patients with mesh infection after tension-free repair of inguinal hernia at the Department of Hernia and Abdominal Wall Surgery,Beijing Chaoyang Hospital from 2018 to 2020 were retrospectively analyzed.Results:The most frequent type of repair was plug implantation, accounting for 57.5% of the procedures. The most common clinical presentation was a chronic sinus. 79.3% patients had a >3-month history of chronic infection. Staphylococcus aureus was the most common bacteria. All patients underwent open debridement. Fifty-one patients had a complete removal of the infected mesh, and 36 had partial removal. All patients were followed up for 18.7-54.2 months. There was no significant difference in the incidence of wound infection, seroma, hematoma, inguinal hernia recurrence, and chronic pain between those with complete removal and that partial removal (all P>0.05). Seventeen cases suffered recurrent sinus in the partial mesh removal group, and the incidence was significantly higher than that in the complete mesh removal group ( P<0.001). Conclusion:Infected mesh removal is an effective treatment for mesh infection after tension-free repair of inguinal hernia and should be removed as completely as possible.

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 investigate the surgical treatment of mesh infection after lapa-roscopic inguinal hernia repair.Methods:The retrospective and descriptive study was conducted. The clinical data of 32 patients with mesh infection after laparoscopic inguinal hernia repair who were admitted to the Affiliated Beijing Chaoyang Hospital of Capital Medical University from January 2018 to December 2020 were collected. There were 30 males and 2 females, aged 59(range, 19-81)years. All patients underwent debridement. Observation indicators: (1) surgical and postopera-tive situations; (2) follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers or percentages, and comparison between was conducted using Fisher exact probability. Results:(1) Surgical and postoperative situations. All 32 patients underwent debridement success-fully, with the operation time and volume of intraoperative blood loss as 110(range, 45-220)minutes and 24(range, 5-200)mL. Of the 32 patients, there were 6 cases undergoing open debridement, 26 cases undergoing laparoscopic exploration with 12 cases clarified the cause of infection, in which 4 cases had intestinal fistula by mesh infection breaking through the peritoneum and invading the intestinal tract and underwent laparoscopic intestinal fistula repair, 3 cases had significant amount of residual medical glue, 3 cases had mesh wrinkling and curling, 2 cases underwent peritoneum suturing with black silk thread. There were 18 cases undergoing complete removal of infected mesh and 14 cases undergoing partial mesh removal. There were 18 cases undergoing removal of infected mesh by laparoscopic posterior approach and 14 cases undergoing removal of infected mesh by open anterior approach. Twenty-nine of the 32 patients had bacterial culture and drug allergy testing, including 24 cases with positive cultures, while 5 cases with negative cultures. The duration of hospital stay of the 32 patients was 27.0(range, 9.0-85.0)days. (2) Follow-up. All 32 patients were followed up for 37.9(range, 18.7-52.5)months. There were 5 cases and 2 cases with postoperative incision infection in the 18 patients undergoing complete removal of infected mesh and 14 cases undergoing partial mesh removal, respectively, showing no significant difference between them ( P>0.05). There were 7 cases, 4 cases with seroma, recurrent sinus in the 18 patients undergoing complete removal of infected mesh and 5 cases, 6 cases in the 14 cases undergoing partial mesh removal, respectively, showing no significant difference between them ( P>0.05). None of patient in the two groups had hematoma, recurrent inguinal hernia or chronic pain. The 7 patients with incision infec-tion recovered effectively after symptomatic treatment. The 12 patients with seroma received no special treatment. The 10 patients with recurrent sinus underwent debridement, in which 9 cases did not experience any further abdominal wall infections and 1 case underwent the third debride-ment. Conclusion:The management of mesh infection following laparoscopic inguinal hernia repair necessitates the expertise of hernia specialists to complete remove the infected mesh or partial remove the infected mesh based on the specific clinical scenario.

The massive accumulation of β-amyloid （Aβ） in the brain is believed to be the first pathological mechanism of Alzheimer's disease （AD）， and the accumulation is mainly resulted from the overproduction and dysfunction in the clearance. Extensive and in-depth research has been carried out on AD. In addition to the drugs which are commonly used in clinical settings to improve cognitive function， Aβ monoclonal antibody aducanumab （Aduhelm） has been successfully marketed in the US， which may delay the progress of AD. Thus， it is a potential method for the treatment of AD to target Aβ， but it is expensive， with many adverse reactions and contraindications， which hinders the clinical promotion. Traditional Chinese medicine， featuring multiple components， multiple targets， multiple pathways， and high safety， can regulate the level of Aβ deposition in the brain， alleviate neurotoxicity， and prevent and treat AD by inhibiting the production and aggregation of Aβ and promoting the clearance in the brain. Berberine， gallic acid， osthole， scutellaria barbata flavonoids， Huannao Yicong decoction and Ditantang can promote α-secretase and inhibit the activity and expression of β- and γ-secretase， thus reducing production of Aβ. Baicalein， aloe-emodin， gallic acid， and curcumin can suppress the aggregation of Aβ， promote its depolymerization， and reduce the toxic effect of Aβ on nerve cells by interacting with the hydrophobic structure of Aβ and the H bond， salt bridge， and β-sheet that mediate the aggregation of Aβ. Curcumin and resveratrol can promote the expression of triggering receptor 2 in bone marrow cells of microglia and the migration and phagocytosis of Aβ in microglia. Bilobalide， Kaixinsan and curcumin can up-regulate the expression of encephalin-degrading enzyme and insulin degrading enzyme to promote Aβ degradation， and geniposide， dihydrotanshinone， dihydroartemisinin， and curcumin can degrade Aβ in cells by activating normal autophagy or inhibiting abnormal autophagy. Cycloastragenol， Danggui Shaoyaosan， Yizhi Fangdai formula and Linggui Zhugan decoction can promote the outflow of Aβ and inhibit the inflow of Aβ by improving the integrity and permeability of the blood-brain barrier （BBB）. Yizhi Fangdai formula and Xueshuantong can promote the polarization of aquaporin 4（AQP4）， allow Aβ to be cleared through the lymphatic system， and reduce the aggregation of Aβ in the brain， thereby relieving or preventing nerve cell damage and improving cognitive function. The above summary aims to provide more sufficient evidence and ideas for the clinical treatment of AD with traditional Chinese medicine.

OBJECTIVETo investigate the relationship between polymorphisms of the growth arrest specific 6 (GAS6) gene and severe preeclampsia in a South West Han Chinese population.

METHODSBlood samples from 167 patients with severe preeclampsia and 312 normal pregnant women as controls from Han Chinese in Chengdu area were analyzed by polymerase chain reaction-restriction fragment length polymorphisms.

RESULTSC and T allele frequencies for +1332C/T site were 85.63% and 14.37% in the patient group, respectively, and 78.04% and 21.96% in control group, respectively. The TT genotype and variant T allelic frequencies of the +1332C/T polymorphism were significantly lower in patients with severe preeclampsia than in the control group (both P<0.05), and the odds ratio for the risk of severe preeclampsia was 0.602 (95%CI: 0.401-0.904) in carriers for the variant T allele (χ=6.045, P=0.014). G and A allele frequencies for 834+7G/A site were 72.75% and 27.25% in case group, respectively, and 74.36% and 25.64% in control group, respectively. The genotype and allele frequencies of the 834+7G/A polymorphism in patients with severe preeclampsia and controls showed no significant differences (both P>0.05). In addition, there was no significant association between the polymorphisms and blood pressure levels in the patient or control groups.

CONCLUSIONThe variant GAS6+1332 T allele is associated with a decreased risk for severe preeclampsia in a South West Han Chinese population. On the other hand, the 834+7G/A polymorphism has no effect on the severe preeclampsia.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; ethnology ; genetics ; Genotype ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Pre-Eclampsia ; ethnology ; genetics ; pathology ; Pregnancy ; Risk Factors ; Severity of Illness Index ; Young Adult

This research is to construct a basic medical sciences’experimental teaching system in order to cultivate innovative talents. It is guided by cultivating innovative practical ability and post compe-tence and implements a teaching mode with “five combinations”by integrating teaching resources, strength-ening interdisciplinary combination , integrating curriculum and organ systems , and optimizing teaching modules and experiment content. A preliminary personnel training mode and experimental teaching system have been constructed for innovative talent cultivation, and correspondently a diversified experiment exami-nation system and teaching quality monitoring system have been constructed through teaching practice, which aims at continuously improving experiment teaching quality and talent training quality.