Objective:To analyze the application effect of nano-carbon lymphatic tracer technology in laparoscopic colon cancer (CC) radical resection based on propensity matching.Methods:Retrospective case-control study was performed in this study. From January 2016 to April 2021, 714 cases of CC patients who underwent laparoscopic CC radical resection in Kunshan Second People′s Hospital were divided into groups according to whether or not the nano-carbon lymphatic tracing technique was applied. Seventy-eight cases in group A were applied with nano-carbon lymphatic tracing technique, while 636 cases in group B were not applied with nano-carbon lymphatic tracing technique. The initial data were matched 1∶3 by the propensity score matching method, and finally group A (73 cases) and group B (219 cases) were obtained. The detection of lymph nodes in the two groups after propensity score matching was compared.Results:By comparing the baseline data of the two groups after propensity score matching, it was found that there were no significant differences in gender, height, weight, body mass index, tumor T stage, tumor N stage, tumor TNM stage, preoperative chemotherapy, or tumor location ( P>0.05). The total number of lymph nodes in group A was higher than that in group B: (22.24 ± 7.08) pieces vs. (19.03 ± 6.29) pieces, and the difference was statistically significant ( t = 3.66, P<0.05); the number of positive lymph nodes and the degree of lymph node metastasis in group A were not significantly different from those in group B ( P>0.05). Tumor T stage T 3, tumor N stage N 0, tumor TNM stage Ⅱ, and preoperative chemotherapy, the total number of lymph nodes in group A was higher than that in group B: 23 (6, 60) pieces vs. 19 (4, 54) pieces , 20 (3, 62) pieces vs. 18 (3, 75) pieces, 23 (6, 59) pieces vs. 20 (7, 54) pieces, 22 (5, 45) pieces vs. 14 (4, 46) pieces, and the difference was statistically significant ( Z = 2.43, 2.70, 2.64 and 3.32; P<0.05); the number of positive lymph nodes and the degree of lymph node metastasis of tumor N stage N 2 in group A were lower than those in group B: 4 (4, 9) pieces vs. 6 (4, 25) pieces , 16 (10, 42) pieces vs. 32 (19, 100) pieces, and the difference between groups was statistically significant ( Z = -2.53 and -2.87, P<0.05). Followed up to April 2022, among the 292 patients, 5 were lost to follow-up, the 3-year disease-free survival rates of 72 patients in group A and 215 patients in group B were 83.33% (60/72) and 91.16% (196/215) respectively, there was no significant difference between two groups ( P>0.05). Conclusions:The number of lymph nodes detected in laparoscopic CC radical resection increases after the application of nano-carbon lymphatic tracing technology.

Objective:To obtain purified protein antigen of guertu virus (GTV) nucleoprotein (NP) and establish a rapid and accurate enzyme-linked immunosorbent assay (ELISA) method for detection of GTV antibody.Methods:Codon optimized GTV NP encoding genes were synthesized, cloned into the pet32a (+) vector, and recombinant expression plasmids were constructed and transformed into BL21 (DE3). Recombinant protein (rNP) obtained from the optimized expression were purified over a Ni column and identified by SDS-PAGE and Western blot. The purified protein was used as the antigen to optimize the reaction conditions, and an indirect ELISA assay for GTV IgG antibody was developed and optimized, which was evaluated and initially applied.Results:The prokaryotic expression plasmid pet32a-NP was successfully constructed, the recombinant protein was highly expressed in E. coli in the form of inclusion bodies, the size was about 44 kD, and the results of Western blot indicated that the recombinant protein had good antigenicity with GTV positive serum. The optimized ELISA (GTV-rNP-iELISA) established in this study showed strong specificity, high sensitivity, and the coefficient of variation within and between batches is less than 10%, and has good repeatability; the detection results are consistent with the IFA detection results. Using the established ELISA method to detect 162 sheep sera from some regions of Xinjiang in 2017-2019, the total positive rate of antibodies was 39.8%.Conclusions:The GTV NP antibody detection ELISA method has good sensitivity, reproducibility, and specificity and has the potential to be a powerful tool for the diagnosis and serological investigation of GTV infection.

Objective:To obtain purified protein antigen of guertu virus (GTV) nucleoprotein (NP) and establish a rapid and accurate enzyme-linked immunosorbent assay (ELISA) method for detection of GTV antibody.Methods:Codon optimized GTV NP encoding genes were synthesized, cloned into the pet32a (+) vector, and recombinant expression plasmids were constructed and transformed into BL21 (DE3). Recombinant protein (rNP) obtained from the optimized expression were purified over a Ni column and identified by SDS-PAGE and Western blot. The purified protein was used as the antigen to optimize the reaction conditions, and an indirect ELISA assay for GTV IgG antibody was developed and optimized, which was evaluated and initially applied.Results:The prokaryotic expression plasmid pet32a-NP was successfully constructed, the recombinant protein was highly expressed in E. coli in the form of inclusion bodies, the size was about 44 kD, and the results of Western blot indicated that the recombinant protein had good antigenicity with GTV positive serum. The optimized ELISA (GTV-rNP-iELISA) established in this study showed strong specificity, high sensitivity, and the coefficient of variation within and between batches is less than 10%, and has good repeatability; the detection results are consistent with the IFA detection results. Using the established ELISA method to detect 162 sheep sera from some regions of Xinjiang in 2017-2019, the total positive rate of antibodies was 39.8%.Conclusions:The GTV NP antibody detection ELISA method has good sensitivity, reproducibility, and specificity and has the potential to be a powerful tool for the diagnosis and serological investigation of GTV infection.

Objective:To express truncated glycoprotein (Gn, Gn1, Gn2, Gn3, Gc1 and Gc2) of Guertu virus (GTV) in Escherichia coli ( E. coli) cells, and prepare polyclonal antibodies against recombinant proteins Gn-His, Gc1-His and Gc2-His after purification. Methods:Gene fragments encoding Gn, Gn1, Gn2, Gn3, Gc1 and Gc2 of GTV DXM strain were amplified by RT-PCR, and cloned into the prokaryotic expression vector pET-32a (+ ) to construct recombinant expression plasmids. The transformed E. coli BL21(DE3) strains carrying expression plasmids were induced by IPTG to express target proteins, which were identified by SDS-PAGE. Recombinant proteins Gn-His, Gc1-His and Gc2-His were purified by nickel affinity chromatography and detected by Western blot using GTV-positive sheep serum for analysis of their antigenicity. New Zealand white rabbits were immunized with the purified recombinant proteins. The titers and specificity of serum antibodies were analyzed by ELISA. Meanwhile, eukaryotic expression vectors pcDNA3.1-Gn, pcDNA3.1-Gc1/Gc2 were constructed and transfected into mammalian Vero cells to evaluate the binding activity of rabbit polyclonal antibodies by indirect immunofluorescence method. The specific reactivity of serum antibodies to recombinant proteins was detected by Western blot. Results:Restriction enzyme analysis and DNA sequencing confirmed that the recombinant expression vectors of pET-32a-Gn, pET-32a-Gn1/Gn2/Gn3, pET-32a-Gc1/Gc2, pcDNA3.1-Gn and pcDNA3.1-Gc1/Gc2 were constructed successfully. The relative molecular mass ( Mr) of the expressed recombinant proteins Gn-His, Gn1/Gn2/Gn3-His, Gc1/Gc2-His were approximately 63.4×10 3, 37.1×10 3, 31.9×10 3, 30.8×10 3, 40×10 3 and 54.4×10 3, respectively. The recombinant proteins could be recognized by GTV-positive sheep serum. The titers of polyclonal antibodies against GTV Gn, Gc1 and Gc2 were 1∶409 600, 1∶204 800 and 1∶6 400, respectively. Indirect immunofluorescence assay and Western blot showed that the prepared rabbit polyclonal antibodies could specifically react with the proteins expressed in eukaryotic cells and the recombinant proteins. Conclusions:The recombinant GTV glycoproteins Gn-His and Gc1/Gc2-His were efficiently expressed and purified and characterized with good immunity. The prepared polyclonal antibodies had high titers and good specificity. This study provided reference for further studying the biological function and detection methods of GTV glycoproteins and research on vaccines.

Objective To establish a model of deep venous thrombosis (DVT) in rabbits. Methods Animal models of venous thrombosis were made by blocking venous flow with a vascular clamp temporarily, injuring the vascular wall, and injecting thrombin in the distal vein in one side of rabbits. Then fixed the hip and knee joints of the operated sides in the flection position with plaster. 48 h later, the femoral veins on both sides were examined with the ultrasonography and pathology. Results All the rabbits survived after operation. The ultrasonography showed that the femoral veins on both sides were virtually anechoic. However, the veins on the operated sides couldn't be compressed and no flow was detected, the control sides were just the reverse. The veins on the operated side were filled with thrombus which had not adhered on the wall, but no thrombosis occurred in the control side. Conclusion A model of DVT was established in rabbits.

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.