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

"The Expert Group on Tumor Ablation Therapy of Chinese Medical Doctor Association, The Tumor Ablation Committee of Chinese College of Interventionalists, The Society of Tumor Ablation Therapy of Chinese Anti-Cancer Association and The Ablation Expert Committee of the Chinese Society of Clinical Oncology" have organized multidisciplinary experts to formulate the consensus for thermal ablation of pulmonary subsolid nodules or ground-glass nodule (GGN). The expert consensus reviews current literatures and provides clinical practices for thermal ablation of GGN. The main contents include: (1) clinical evaluation of GGN, (2) procedures, indications, contraindications, outcomes evaluation and related complications of thermal ablation for GGN and (3) future development directions.
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Objective:To investigate the number of necroptotic renal tubular epithelial cells in renal tissues of patients with chronic kidney disease (CKD) and the correlation with clinicopathologic parameters, and explore its role in the progression of the excessive loss of renal tubular cells and chronic kidney injury.Methods:Renal tissue samples from 60 patients (18-65 years old) with CKD proven by kidney biopsy in the First Affiliated Hospital of Hainan Medical University from June 2017 to June 2019 were collected. According to internationally accepted K/DOQI guidelines, the patients were divided into 1-4 stages of CKD, with 15 cases in each stage. The number of necroptotic renal tubular epithelial cells in patients with different stages of CKD was detected using receptor-interacting protein 3 (RIP3) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) fluorescent staining, and the expression of RIP3 and MLKL, marker protein of necroptosis, was detected by immunohistochemistry. Pearson correlation analysis was used to analyze the correlation between the percentage of necroptotic renal tubular epithelial cells and clinicopathologic parameters. In addition, the expression of angiotensinogen Ⅱ receptor (AT2R) in renal tissue and its correlation with the percentage of necroptotic renal tubular epithelial cells were analyzed.Results:With the development of CKD, the structural destruction of renal tubules in patients with CKD was gradually aggravated, and the renal tubules in the corresponding areas were atrophied, accompanied by worsening interstitial fibrosis. The adjacent renal tubules were focally dilated and numerous protein tubules were seen in the tubules. Importantly, renal tubular injury score in second and third stage of CKD was significantly higher than that in control group (both P<0.01). TUNEL+RIP3 immunofluorescence staining results showed that the percentage of TUNEL/RIP3 double positive renal tubular epithelial cells (necroptotic renal tubular epithelial cells) in renal tubules of the second and third stage of CKD was higher (all P<0.01). Immunohistochemical results showed that RIP3, MLKL and AT2R proteins were mainly expressed in cytoplasm of renal tubular epithelial cells, and the expression of RIP3, MLKL and AT2R in renal tubular epithelial cells was higher in the second and third stage of CKD patients (all P<0.05). Pearson correlation analysis showed that the percentage of necroptotic renal tubular epithelial cells was positively correlated with blood urea nitrogen ( r=0.514, P=0.003), serum creatinine ( r=0.507, P=0.019), serum cystatin C ( r=0.571, P=0.026), serum uric acid ( r=0.592, P=0.008), renal tubules injury score ( r=0.901, P<0.001), renal interstitial fibrosis index ( r=0.700, P=0.001) and the expression of AT2R protein in renal tissue ( r=0.715, P=0.001). Conclusions:As CKD progresses, necroptosis of renal tubular epithelial cells in CKD patients occurs. The necroptotic cell death may be an important factor leading to renal tubular epithelial cell excessive death and the progression of chronic kidney injury. Furthermore, necroptosis of renal tubular epithelial cells may be related to the high expression of AT2R in kidney tissue.

Objective To investigate the effect of locking compression plate(LCP)and anatomi-cal plate(AP)in the treatment of closed tibial Pilon fracture and its influence on lower limb functional re-habilitation.Methods A total of 82 patients with closed tibial Pilon fractures were randomly divided into LCP internal fixation group(LCP group,n=41)and AP internal fixation group(AP group,n=41).The perioperative indexes and the incidence of postoperative complications were compared between the two groups.The ankle function was evaluated by American Orthopaedic Foot & Ankle Society Ankle Hindfoot Scale(AOFAS-AHS),the lower limb function was evaluated by Lysholm scale,the serum levels of inter-leukin-1β(IL-1β)and interleukin-6(IL-6)were detected.Results The intraoperative blood loss,opera-tion time,hospitalization time,first ground time,fracture healing time were(103. 34 ± 11. 34)ml, (47.65 ± 7.89)min,(9.01 ± 2.23)d,(5.31 ± 1.27)d,(16.23 ± 2.12)weeks in LCP group respective-ly,the AP group were(132.25 ± 34.41)ml,(60.54 ± 11.23)ml,(11.43 ± 2.57)d,(6.23 ± 1.56)d, (23.12 ± 3.31)weeks,respectively.The LCP group were significantly lower than AP group(P<0.05);The excellent rate of ankle function of LCP group was 95.12%,significantly higher than 82.93% in the AP group(P<0.05);The incidence of postoperative complications was 4.88% in the LCP group,which was significantly lower than 29.27% in the AP group(P<0.05);At 3,6 and 12 months after operation, the AOFAS-AHS scores in the LCP group were(69.52 ± 4.18)points,(78.89 ± 6.73)points and (87.23 ± 6.34)points respectively,the AP group were(65.09 ± 4.45)points,(70.13 ± 5.34)points and (76.69 ± 5.91)points respectively,the LCP group were significantly higher than AP group(P<0.05);At 3,6 and 12 months after operation,the Lysholm scores were(77.12 ± 6.43)points,(82.12 ± 7.81)points and(86.19 ± 8.11)points in LCP group,AP group were(67.25 ± 5.56)points,(72.21 ± 7.23)and (77.12 ± 7.54)points,the LCP group was significantly higher than AP group(P<0.05).At 3 d and 4 weeks after operation,the serum levels of IL-1β in LCP group were(0.37 ± 0.09)pg/ml,(0.19 ± 0.06) pg/ml,while in AP group were(0.45 ± 0.13)pg/ml,(0.27 ± 0.09)pg/ml;the serum levels of IL-6 in LCP group were(201.23 ± 30.12)ng/L,(102.23 ± 25.21)ng/L,while in AP group were(246.71 ± 41.23)ng/L,(158.95 ± 25.21)ng/L.The AP group were significantly lower than those in AP group(P<0.05).Conclusion LCP and AP in treatment of closed tibial Pilon fractures have a significant effect,but LCP can reduce the surgical trauma,shorten the operation time and postoperative recovery time,to a cer-tain extent,improve the ankle and lower limb function,and reduce the postoperative fixation fracture heal-ing and other complications.

Objective To evaluate the prevalence of chronic kidney disease (CKD) in Chinese adult health check-up population,and to compare with the prevalence of CKD in the study of the general population as well as the large CKD cross-sectional study in China.Methods Epidemiological studies about CKD in Chinese adults health check-up population from January 2007 to December 2017 were searched in PubMed,SinoMed,CNKI,VIP and Wanfang Data.Meta-analysis of the prevalence of CKD was performed with software of Stata 12.0.Subgroup analyses of CKD staging,urban and rural,as well as geographical areas of the general population were executed.Results Twenty-two studies from adult health check-up population were included (238 349 persons).Egger's regression showed no publication bias (P ＞ 0.05).The unstandardized prevalence rate of CKD was 12.49％ (male 12.8％,female 12.5％).The respective unstandardized prevalences of proteinuria,hematuria and eGFR decline were 5.90％,5.83％ and 2.75％.The unstandardized prevalences of CKD in urban and rural population were 13.21％ and 11.90％.The stages of CKD were mainly concentrated in the early stages.There was no significant difference in the non-standard detection rate of total eGFR decline among the adult medical examination population,the general population and the population studied cross-sectionally (P ＞ 0.05).Furthermore,no significant difference in the non-standard detection rate of total hematuria and male hematuria was found between the adult health check-up population and the general population.In addition,the total proteinuric non-standard detection rate of the adult general population was similar with that of population studied cross-sectionally (P ＞ 0.05).Conclusions The prevalence of CKD in Chinese adults is higher,the overall prevalence is however underestimated.The results of epidemiological investigation in adult health check-up population are similar to those of the general population,especially in men.

Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirusAd-Apoptin-hTERTp-E1A(ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancerA549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods:ATV was used to infectA549-luc cells andA549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode ofATV. Results: WST-1 and crystal violet staining showed thatATV had significant inhibitory effect on bothA549-luc and A549 cells ( P <0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h ( P <0.05 or P <0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI. Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set. The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h( P <0.05 or P <0.01). Conclusion: Insertion of luciferase didn’t significantly change the inhibitory effect and inhibitory mode ofATV onA549-luc cells.ATV exerted its in vitro inhibitory effect onA549-luc and A549 cells by inducing cell apoptosis.

Objective To discuss the clinical application of preoperative tumor-feeding artery emboli -zation in treating hypervascular giant meningiomas.Methods A total of 71 patients with giant meningioma (maximum diameter ＞5 cm),who were admitted to authors' hospital during the period from April 2013 to August 2014,were selected for this study.Preoperative MRI demonstrated that the lesions showed obvious enhancement with rich blood supply.The patients were divided into the study group (using preoperative embolization,n =38) and the control group (not using preoperative embolization,n =33).Based on preoperative DSA findings,the lesions were classified into type Ⅰ and type Ⅱ.Tumor-feeding artery embolization with PAV particles (200-300 μm) was employed in the patients of the study group,while only cerebral angiography was adopted in the patients of the control group.The amount of intraoperative blood loss,tumor resection time and embolization complications in both groups were recorded.Postoperative MRI scan was performed to observe the degree of tumor resection.Results The amount of intraoperative blood loss and the tumor resection time in the study group were (562±178) ml and (5.45±2.13) h respectively,which in the control group were (833±234) ml and (7.23±2.45) h respectively;the differences between the two groups were statistically significant (both P＜0.05).The tumor resection degree of Simpson classification Ⅲ and ＜ Ⅲ was obtained in 27 patients of the study group (27/38,71.0％) and in 19 patients of the control group (19/33,57.6％),the difference between the two groups was statistically significant (P＜0.05).After embolization,cerebral edema became aggravated in 7 patients,ischemic necrosis of the scalp was observed in 2 patients,and one patient developed epileptic seizure during the operation.After symptomatic treatment,all the above complications were relieved.Conclusion For the treatment of hypervascular giant meningiomas,preoperative tumor-feeding artery embolization can strikingly reduce the amount of intraoperative blood loss,shorten the operation time,and improve the tumor resection degree,therefore,this therapy is worthy of clinical promotion and application.

Background and purpose:Since the number and tumor size of localized liver metastases can be controlled, local minimally invasive treatment can improve the survival of patients. Hence, microwave ablation has become an important treatment method for liver metastases. This study was to investigate the value of percutaneous microwave ablation in the treatment of tumor metastases. Methods: From Sep. 2011 to Oct. 2014, 26 advanced nasopharyngeal carcinoma patients with post-chemotherapy consolidation, liver metastases were collected. All the patients with the number of tumor lesions less than 3, diameter less than 5 cm, no other distant metastases was excluded. The ultrasound-guided percutaneous microwave ablation was used for 26 patients. Finally, 43 ablations were completed followed by liver function test, enhanced CT and MRI diagnosis 1 month later. mRECIST criteria was used to evaluate the effcacy of cancer treatment. Progression-free survival (PFS) and overall survival (OS) were calculated. Results:Twenty-six cases of a total of 53 lesions, including complete ablation (CA) 20 patients (20/26, 77.0%), partial ablation (PA) 3 patients (3/26,11.5%). The overall effciency was 88.5%(CA+PA) with no serious complications. 6 months, 1-, 2-year survival rates of 26 patients were 96.1%, 65.3%and 23.0%. PFS was 11.4 months. The median survival time (MST) was 11.9 months, while OS was 23.7 months. Conclusion:Percutaneous microwave ablation for limited liver metastases of nasopharyngeal carcinoma is a minimally invasive, safe and effective treatment method.

Objective To discuss the clinical application of percutaneous transhepatic cholangiopancreatic drainage (PTCD) combined with CT-guided microwave ablation in treating hepatic hilar cholangiocarcinoma. Methods The clinical data of 29 patients with inoperable Ⅲ and Ⅳ type hilar cholangiocarcinoma complicated by obstructive jaundice, who were admitted to authors’ hospital during the period from December 2012 to August 2014, were retrospectively analyzed. The diagnosis of bile duct adenocarcinoma was confirmed by pathology in all patients. Of the 29 patients, both internal and external biliary tract drainage was employed in 19, external biliary tract drainage in 4, and bilateral (both left and right side) bile duct drainage in 6. CT-guided microwave ablation was carried out when the liver function became improved. A total of 46 procedures of microwave ablation were completed in the 29 patients with a mean of 1.5 times per patient. Hepatic function tests, enhanced CT or MR scan were performed 4-8 weeks after treatment. According to mRECIST criteria the therapeutic results were evaluated; the serum bilirubin levels were recorded; the disease progress and the patient’s survival time were followed up. Results One month after the treatment, complete response (CR) was obtained in 15 patients (15/29, 51.7%), and partial remission (PR) in 17 patients (17/29, 58.6%), with the overall efficacy (CR﹢PR) being 82%. After the treatment, one patient developed hepatic metastasis and another one had pulmonary metastasis. Postoperative serum bilirubin levels showed an obvious decrease in all 29 patients. The 6-month, one-year and two-year survival rates were 68.9% (20/29), 31.0% (9/29) and 6.8% (2/29) respectively; the median survival time was 8.9 months and the overall survival time was 11.7 months. Conclusion For the treatment of hepatic hilar cholangiocarcinoma, PTCD combined with CT-guided microwave ablation is minimally invasive, clinically safe and effective.

Informed consent right is not just for basic ethical consideration, but is important for protect-ing patient’s right by law, which is expressed through informed consent contract. The appraised individ-uals of forensic clinical examination have the similar legal status as the patients in medical system. However, the law does not require informed consent right for the appraised individuals. I recommend giving certain informed consent right to the appraised individuals in the forensic clinical examination. Under the contracted relationship with the institution, the appraised individuals could participate in the examination process, know the necessary information, and make a selected consent on the examination results, which can assure the justice and fairness of judicial examination procedure.