To establish a rapid visual detection method for Akabane virus(AKAV)on site,specific primers and probes based on the S fragment of AKAV were designed in this experiment.Corre-sponding groups were added to the primers or probes to fulfil the requirement of the combination of recombinase polymerase amplification(RPA)with lateral flow dipstick(LFD).The reaction temperature and time,concentrations of the primer and probe were optimized to establish the RPA-LFD method for detecting AKAV.After that,the specificity,sensitivity and clinical reliability of the method were evaluated.The results showed that after 20 minutes of reaction at 37 ℃,the test results could be read on LFD paper.There was no cross reaction against blue tongue virus,Pasteurella multocida,bovine infectious rhinotracheitis virus and bovine Mycoplasma bovis,and the detection limit was 2.5 × 100 copies/μL of standard plasmid.Detection of clinical samples showed a consistent results with that by RT-PCR method.These findings indicated that the RPA-LFD method established had the advantages of good specificity,high sensitivity,simple operation and visualization,and could be applied to clinical detection,which provides new technical support for the rapid diagnosis and prevention and control of AKAV.

Objective To investigate the characteristics of KRAS,NRAS,BRAF,and PIK3CA mutations in different colorectal cancer(CRC)metastatic sites and their correlation with prognosis.Methods The mutations of KRAS,NRAS,BRAF,and PIK3CA were detected in 205 cases by using a mutation amplification refractory mutation system(ARMS),and their correlation with clinical fea-tures,metastatic sites,and related prognosis was analyzed.Results The mutation rates of KRAS,NRAS,BRAF,and PIK3CA were 57.1％,4.4％,4.4％and 3.9％,respectively.Mutations of KRAS are increased in women,patients with colorectal cancer,and patients with stage Ⅲ-Ⅳ colorectal cancer.The BRAF mutation rate in right-sided colon cancer was increased(P＜0.05).KRAS mutations were com-monly seen during pulmonary metastasis,while NRAS mutation rates were higher during liver metastasis,and the proportion of peritoneal metastases was higher when BRAF mutated.KRAS and NRAS mutations were identified as elevated risks for lung and liver metastasis in colorectal cancer(CRC)(P＜0.01).BRAF mutation types lead to shorter overall survival(OS)in CRC cases with liver and peritoneal metastasis,whereas KRAS mutations indicate poorer prognosis in lung metastasis.Both KRAS and BRAF mutations are independent prog-nostic indicators of unfavorable outcomes in CRC with lung and liver metastasis,respectively(P＜0.01).Conclusion KRAS,NRAS and BRAF mutations are closely associated with distant metastasis and prognosis in CRC.Clinical practice should routinely assess these mutations to provide critical guidance for treatment decisions.

Microbial denitrification is a major pathway for nitrogen removal from water bodies. However, denitrification is often difficult to continue when there is a lack of microbially available organic matter in the water body to serve as electron donors. In recent years, studies have shown that some denitrifying bacteria can directly utilize photoelectrons generated by sunlight-excited semiconductor minerals or natural organic matter for denitrification without the need for bioavailable organic matter as electron donors. This process is defined as microbial photoelectrotrophic denitrification. The discovery of microbial photoelectrotrophic denitrification phenomenon reshapes the previous knowledge about the chemoheterotrophic mode of denitrifying bacteria and broadens the pathway of nitrogen removal by the new photoelectrotrophic metabolism, which is of great significance to our understanding and exploration of sunlight-driven nitrogen cycling process. In this paper, we comprehensively sort out the existing research reports in the field of microbial photoelectrotrophic denitrification, systematically summarize the principle and the current research progress of microbial photoelectrotrophic denitrification, deeply analyze the problems and challenges faced by this technology, and make an outlook on the future research directions and application prospects of this technology, providing a reference for the further research and application of this technology.
Denitrification/physiology* ; Nitrogen/isolation & purification* ; Bacteria/metabolism* ; Sunlight ; Phototrophic Processes

With the increasing incidence and mortality of cancer, the limitations of conventional therapies have become increasingly apparent, necessitating the development of more precise immunotherapeutic approaches. mRNA vaccines have emerged as a prominent research focus in cancer immunotherapy due to their advantages in antigen design, rapid production, and enhanced safety profiles. This article reviews the advances in molecular design optimization of mRNA cancer vaccines such nucleoside modification, 5’-/3’-untranslated region (UTR) optimization and open reading frame (ORF) engineering to enhance efficacy and stability), key antigen types (viral antigens, tumor-associated antigens, tumor-specific antigens, and immunomodulators), delivery systems, and clinical research. By expressing specific antigens in vivo, these vaccines activate immune recognition and elimination of cancer cells. Currently, multiple mRNA vaccines have entered clinical trials, often in combination with immune checkpoint inhibitors or other immunotherapies. Despite their promising potential, challenges such as mRNA instability, difficulty in targeted delivery, immune tolerance, and tumor immune evasion still exist. Future breakthroughs will require advances in neoantigen prediction, targeted delivery, and combination therapy.

Objective:To investigate the influence of 17β-estradiol(17β-E2)in the proliferation and differentiation capabilities of primary cultured hippocampal neural stem cells(NSCs),and to clarify its potential mechanism.Methods:The NSCs were isolated from the hippocampal tissue of SD rats within 24 h of birth,and divided into control group and 17β-E2 group.Immunofluorescence method was used to detect the expressions of Nestin and 5-ethynyl-2'-deoxyuridine(EdU)in NSCs in two groups,and the proliferation rates of NSCs were calculated.Western blotting method was used to detect the expression level of Nestin protein.Flow cytometry was used to analyze the percentages of NSCs at different cell cycles.Immunofluorescence method was used to identify the expressions of markers for neurons β Ⅲ-tubulin and astrocyte marker glial fibrillary acidic protein(GFAP)in the NSCs after differentiation,and the relative ratio of neurons to astrocytes was calculated.Western blotting method was used to detect the expression levels of β Ⅲ-tubulin and GFAP proteins as well as phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),phosphorylated Akt(p-Akt),glycogen synthase kinase-3 beta(GSK-3β),phosphorylated GSK-3β(p-GSK-3β),and beta-catenin(β-catenin)proteins related to PI3K/Akt/GSK-3β pathway.Results:Immunofluorescence assay revealed positive Nestin expression in NSCs in two groups;compared with control group,the proliferation rate of NSCs in 17β-E2 group was increased(P＜0.01).Compared with control group,the expression level of Nestin protein in the NSCs in 17β-E2 group was increased(P＜0.05),and the percentage of NSCs in S phase was increased(P＜0.01).Compared with control group,the relative ratio of neurons to astrocytes in 17β-E2 group was significantly increased(P＜0.05).After differentiation,compared with control group,the expression level of β Ⅲ-tubulin protein in the NSCs in 17β-E2 group was increased(P＜0.05),and the expression level of GFAP protein was decreased(P＜0.01),while the expression levels of Akt,p-Akt,β-catenin,and p-GSK-3β proteins were increased(P＜0.05 or P＜0.01),and the expression level of GSK-3β protein was decreased(P＜0.05).Conclusion:17β-E2 can promote the proliferation of NSCs and facilitate their differentiation towards neurons,and its mechanism may be related to the PI3K/Akt/GSK-3β signaling pathway.

To establish a rapid visual detection method for Akabane virus(AKAV)on site,specific primers and probes based on the S fragment of AKAV were designed in this experiment.Corre-sponding groups were added to the primers or probes to fulfil the requirement of the combination of recombinase polymerase amplification(RPA)with lateral flow dipstick(LFD).The reaction temperature and time,concentrations of the primer and probe were optimized to establish the RPA-LFD method for detecting AKAV.After that,the specificity,sensitivity and clinical reliability of the method were evaluated.The results showed that after 20 minutes of reaction at 37 ℃,the test results could be read on LFD paper.There was no cross reaction against blue tongue virus,Pasteurella multocida,bovine infectious rhinotracheitis virus and bovine Mycoplasma bovis,and the detection limit was 2.5 × 100 copies/μL of standard plasmid.Detection of clinical samples showed a consistent results with that by RT-PCR method.These findings indicated that the RPA-LFD method established had the advantages of good specificity,high sensitivity,simple operation and visualization,and could be applied to clinical detection,which provides new technical support for the rapid diagnosis and prevention and control of AKAV.

Objective To investigate the characteristics of KRAS,NRAS,BRAF,and PIK3CA mutations in different colorectal cancer(CRC)metastatic sites and their correlation with prognosis.Methods The mutations of KRAS,NRAS,BRAF,and PIK3CA were detected in 205 cases by using a mutation amplification refractory mutation system(ARMS),and their correlation with clinical fea-tures,metastatic sites,and related prognosis was analyzed.Results The mutation rates of KRAS,NRAS,BRAF,and PIK3CA were 57.1％,4.4％,4.4％and 3.9％,respectively.Mutations of KRAS are increased in women,patients with colorectal cancer,and patients with stage Ⅲ-Ⅳ colorectal cancer.The BRAF mutation rate in right-sided colon cancer was increased(P＜0.05).KRAS mutations were com-monly seen during pulmonary metastasis,while NRAS mutation rates were higher during liver metastasis,and the proportion of peritoneal metastases was higher when BRAF mutated.KRAS and NRAS mutations were identified as elevated risks for lung and liver metastasis in colorectal cancer(CRC)(P＜0.01).BRAF mutation types lead to shorter overall survival(OS)in CRC cases with liver and peritoneal metastasis,whereas KRAS mutations indicate poorer prognosis in lung metastasis.Both KRAS and BRAF mutations are independent prog-nostic indicators of unfavorable outcomes in CRC with lung and liver metastasis,respectively(P＜0.01).Conclusion KRAS,NRAS and BRAF mutations are closely associated with distant metastasis and prognosis in CRC.Clinical practice should routinely assess these mutations to provide critical guidance for treatment decisions.

Objective To compare the safety and clinical efficacy of lesion removal combined with percutaneous pedicle screw fixation with classical posterior lesion removal in the treatment of lumbar brucelli spondylitis(LBS)by unilateral biportal endoscopic technique with transforaminal lumbar interbody fusion(UBE-LIF)technique.Methods The clinical data of 32 patients with LBS admitted by the Department of Spine and Orthopedics of Gansu Provincial Hospital of Traditional Chinese Medicine from January 2020 to January 2022 were retrospectively analyzed,and the clinical data of the 32 LBS patients were divided into 15 cases in the UBE-LIF group and 17 cases in the posterior group.The general data,surgery-related indexes,and postoperative pathological HE staining of the two groups were recorded and analyzed.The patients'clinical recovery was assessed according to their erythrocyte sedimentation rate(ESR)and C-reactive protein(CRP),low back pain visual analogue score(VAS),Japanese Orthopaedic Association(JOA)score,and Oswestry Dysfunction Index(ODI)preoperative,1 week after surgery,1,3,6 months and 1 year after surgery.Lumbar lordosis angle(LL)and intervertebral space height(DH)were measured by imaging before surgery and at the last follow-up,and intervertebral bone graft fusion was assessed using Suk grading criteria.Results Both groups successfully completed the operation and no serious postoperative complications occurred.There were no significant differences in gender,age,surgical segment,operation time,preoperative ESR and CRP,preoperative VAS,JOA score and ODI index,preoperative LL and DH(P＞0.05).The intraoperative blood loss,postoperative drainage,postoperative getting out of bed,and postoperative hospital stay in UBE-LIF group were significantly lower than those in the posterior group(P＜0.001).Pathological examination of diseased tissues was performed during surgery,all of which was consistent with brucellosis changes.Patients in both groups were followed up for 12-18 months,with an average of 14.8 months.The VAS,JOA score,and ODI index at all postoperative time points in the two groups were significantly improved compared with the preoperative period(P＜0.05).The difference between the two groups was significantly greater than that in the postoperative group:VAS score was lower in UBE-LIF group than in the posterior group(P＜0.01),CRP in both groups was higher than that in the preoperative group,and the elevation level was significantly lower in UBE-LIF group than in the posterior group(P＜0.001).There was no significant difference in ESR between the two groups compared with that before surgery(P＞0.05).There were no significant differences in VAS,JOA score,ODI index,CRP or ESR between the remaining time points after surgery(P＞0.05).At the last follow-up,imaging examination showed that the overall fusion rate of intervertebral bone graft in UBE-LIF group was 93.3％and 94.1％in the posterior group,without significant difference(x2=0.246,P=0.884).LL and DH were significantly improved in both groups compared with preoperative ones(P＜0.01),and the two groups did not significantly differ before and after surgery(P＞0.05).Conclusion Both surgical treatments for LBS are safe effect.Compared with posterior lesion removal bone graft fusion internal fixation,UBE-LIF technology combined with percutaneous pedicle screw internal fixation has the advantages of clear intraoperative vision,less blood loss,faster early postoperative recovery,and shorter postoperative hospital stay,and thus is a feasible surgical method for the minimally invasive treatment of LBS.

OBJECTIVE To investigate the effects of Codonopsis pilosula polysaccharide （CPP） regulating the nuclear factor- erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway on gastric mucosal injury in rats with chronic atrophic gastritis （CAG）. METHODS Rats were randomly divided into control group， model group， CPP low-dose， medium-dose and high-dose groups （CPP 10， 20， 40 mg/kg）， and ML385 group （Nrf2 inhibitor ML385 30 mg/kg+CPP 40 mg/kg）， 10 rats per group. CAG rat model was established using N-methyl-N′- nitro-N-nitrosoguanidine combined with irregular diet， then they were given drugs for consecutive 6 weeks. HE staining was used to observe the pathological changes in gastric tissue morphology； the levels of serum gastrin （GAS）， motilin （MTL）， pepsin （PP）， as well as tumor necrosis factor-α （TNF-α）， interleukin-8 （IL-8） malondialdehyde （MDA） and superoxide dismutase （SOD） in gastric mucosal tissue were detected； TUNEL assay was used to observe gastric mucosal tissue cell apoptosis； immunohistochemical assay was adopted to observe the expressions of Nrf2 and recombinant Bcl2 associated X protein （Bax） in gastric mucosal tissue； Western blot was used to detect the expressions of Nrf2， HO-1， Bax and Bcl-2 proteins in gastric mucosal tissue. RESULTS Compared with the control group， the gastric mucosal tissue was damaged； the levels of GAS， MTL， PP and SOD， and the protein expressions of Nrf2， HO-1 and Bcl-2 were significantly reduced in model group （P＜0.05）， while the levels of MDA， TNF-α and IL-8， the cell apoptosis index， and the protein expression of Bax were significantly increased （P＜0.05）. Compared with model group， CPP low-dose， medium-dose and high- dose groups showed varying degrees of improvement in gastric mucosal histopathology； the levels of the quantitative indicators were significantly reversed （P＜0.05）. Nrf2 inhibitor ML385 significantly attenuated the improvement effect of high-dose CPP on the above indicators in CAG rats （P＜0.05）. CONCLUSIONS CPP can improve gastric mucosal injury in CAG rats， and inhibit oxidative stress， inflammatory response， and cell apoptosis. The mechanism of action may be related to the activation of Nrf2/HO-1 signaling pathway.

Currently, obesity and its complications have become increasingly serious health issues. Bariatric surgery is an effective method of treating obesity and related metabolic complications. Among them, Roux-en-Y gastric bypass (RYGB) is still considered the “gold standard” procedure for bariatric surgery. Small bowel obstruction is one of the possible complications after RYGB, and in addition to the formation of intra-abdominal hernias, kinking of the jejunojejunal anastomosis is an important cause of small bowel obstruction. The early clinical symptoms of kinking of the jejunojejunal anastomosis often lack clarity in the early stages. Therefore, early diagnosis, prevention, and effective treatment of kinking of the jejunojejunal anastomosis are challenging but crucial. The occurrence of kinking of the jejunojejunal anastomosis may be related to surgical techniques and the surgeon′s experience. The use of anti-obstruction stitch, mesenteric division, and bidirectional jejunojejunal anastomosis may be beneficial in preventing kinking of the jejunojejunal anastomosis. If kinking of the jejunojejunal anastomosis occurs, timely abdominal CT scans and endoscopic examinations should be performed. Gastric and intestinal decompression should be initiated immediately, and exploratory surgery should be prepared.