Objective To prepare mouse monoclonal antibodies against the ectodomain of E2 (E2ecto) glycoprotein of Western equine encephalitis virus (WEEV). Methods A prokaryotic expression plasmid pET-28a-WEEV E2ecto was constructed and transformed into BL21 (DE3) competent cells. E2ecto protein was expressed by IPTG induction and presented mainly as inclusion bodies. Then the purified E2ecto protein was prepared by denaturation, renaturation and ultrafiltration. BALB/c mice were immunized with the formulated E2ecto protein using QuickAntibody-Mouse5W as an adjuvant via intramuscular route, boosted once at an interval of 21 days. At 35 days post-immunization, mice with antibody titer above 1×10⁴ were inoculated with E2ecto intraperitoneally, and spleen cells were fused with SP2/0 cells three days later. Hybridoma cells secreting specific monoclonal antibodies were screened by the limited dilution method, and ascites were prepared after intraperitoneal inoculation of hybridoma cells. The subtypes and titers of the antibodies in ascites were assayed by ELISA. The biological activity of the mAb was identified by immunofluorescence assay(IFA) on BHK-21 cells which were transfected with eukaryotic expression plasmid pCAGGS-WEEV-CE3E2E1. The specificity of the antibodies were evaluated with E2ecto proteins from EEEV and VEEV. Results Purified WEEV E2ecto protein was successfully expressed and obtained. Four monoclonal antibodies, 3G6G10, 3D7G2, 3B9E8 and 3D5B7, were prepared, and their subtypes were IgG2c(κ), IgM(κ), IgM(κ) and IgG1(κ), respectively. The titers of ascites antibodies 3G6G10, 3B9E8 and 3D7G2 were 10⁵, and 3D5B7 reached 10⁷. None of the four antibody strains cross-reacted with other encephalitis alphavirus such as VEEV and EEEV. Conclusion Four strains of mouse mAb specifically binding WEEV E2ecto are successfully prepared.
Horses ; Animals ; Mice ; Encephalitis Virus, Western Equine ; Ascites ; Immunosuppressive Agents ; Antibodies, Monoclonal ; Immunoglobulin M

Objective To investigate the genetic causes of auditory neuropathy with optic atrophy in a family.Methods The proband's medical history and family history were inquired in detail,and relevant clinical examina-tions were performed to confirm the diagnosis of auditory neuropathy with optic atrophy,and the genetic pedigree of the family was drawn.Peripheral blood of proband(Ⅲ-7)was collected for whole exome sequencing,and the patho-genicity of the detected mutations were interpreted.Blood samples of proband's wife(Ⅲ-8),eldest daughter(Ⅳ-7),second daughter(Ⅳ-9)and son(Ⅳ-10)were tested for mutation sites by Sanger sequencing.Combined with clinical manifestations and examination results,the family was studied.Results The genetic pattern of this family was autosomal dominant.The proband showed decreased visual acuity at the age of 19,bilateral sensorineural deaf-ness at the age of 30,and decreased speech recognition rate.Among 20 members of the family of 5 generations,10(2 deceased)showed similar symptoms of hearing and visual impairment.Proband(Ⅲ-7),eldest daughter(Ⅳ-7)and son(Ⅳ-10)underwent relevant examination.Pure tone audiometry showed bilateral sensorineural deafness.ABR showed no response bilaterally.The 40 Hz AERP showed no response in both ears.OAE showed responses in some or all of the frequencies.No stapedial reflex was detected.The eye movement of Ⅲ-7 and Ⅳ-10 were reasona-ble in all directions,and color vision was normal.Ocular papilla atrophy was observed in different degrees in fundus examination.OCT showed thinning of optic disc nerve fibers in both eyes,and visual evoked potential showed pro-longed P100 wave peak.They were diagnosed as hereditary auditory neuropathy with optic atrophy.A mutation of the OPA1 gene c.1334G＞A(p.Arg445His,NM_015560.2)at a pathogenic locus on chromosome 3 was detected by whole exon detection in Ⅲ-7.The results of generation sequencing analysis showed that the OPA1 gene c.1334G＞A(p.Arg445His,NM_015560.2)mutation of chromosome 3 was also found in Ⅳ-7 and Ⅳ-10.Meanwhile,the gen-otypes of Ⅲ-8 and Ⅳ-9 were wild homozygous,that is,no mutation occurred.Conclusion The OPA1 c.1334G＞A(p.Arg445His,NM_015560.2)mutation site might be the pathogenic mutation in this family.

Objective To investigate the effect and safety of common electric knife clamp coagulation technique in rabbit thyroidectomy.Methods According to the random number table method,12 New Zealand rabbits were divided into the clamp coagulation group and the ultrasonic scalpel group,with 6 rabbits in each group.The middle part of the thyroid gland in the clamp coagulation group was severed by common electric knife clamp coagulation,while that in the ultrasonic scalpel group was severed by ultrasonic scalpel electrocoagulation.The postoperative conditions of rabbits in the two groups were observ.The severed thyroid tissue was stained with hematoxylin-eosin(HE),and its histopathology after thermal damage was observed under the light microscope.The scope of thermal damage was determined.On the 1st,3rd and 7th day after operation,the auricular venous blood of all rabbits was collected to assess the serum interleukin-6(IL-6)and C-reactive protein(CRP)levels by enzyme-linked immunosorbent assay.The rabbits were killed on the 7th day after operation,and the residual thyroid glands were removed and stained by HE.The pathological changes and inflammatory cell infiltration were observed under the light microscope.Results The rabbits in the 2 groups survived well after operation,and the operative area healed well.No obvious effusion,blood clot,bleeding,incision infection or other complications were found in the residual cavity.Under the light microscope,the surface of the incisal margin of the thyroid gland showed obvious lesions.In the injured area,some cell structures were damaged with coagulated necrosis,some follicles were ruptured,and the contents inside were solidly concentrated and deeply stained.The cytoplasmic eosinophils in parafollicular cells increased,and nuclear pyknosis,fragmentation,and even dissolution occurred.The thermal damage ranges of thyroid tissues in the ultrasonic scalpel and clamp coagulation groups were(0.72± 0.10)mm and(0.88±0.11)mm,respectively.The range of thermal damage in the clamp coagulation group was significantly greater than that in the ultrasonic scalpel group(t=-2.740,P＜0.05).On the 1st,3rd and 7th day after surgery,there was no significant difference in the levels of serum CRP and IL-6 between the two groups(P＞0.05).The serum IL-6 levels in both groups on the 3rd and 7th day after surgery were significantly higher than those on the 1st day after surgery(P＜0.05).There was no significant difference in serum IL-6 level on the 3rd and 7rd day after surgery in the two groups(P＞0.05).Thyroid follicular atrophy,glia reduction,follicular epithelial hyperplasia,collagenization and hyperplasia of interstitial fibers were observed in the residual thyroid sections of both groups.No obvious inflammatory cell infiltration was observed.Conclusion In rabbit thyroidectomy,it is safe to remove the thyroid gland using the common electric knife clamp coagulation technique.In terms of preventing thermal damage,the ultrasonic scalpel is better than the common electric knife clamp coagulation technique,but the thermal damage to thyroid tissues caused by the common electric knife clamp coagulation technique is within the safe operating range.

Lyme disease is a natural zoonotic infectious disease transmitted by ticks infected by different genotypes of Borre-lia burgdorferi sensu lato,which was discovered in the 1970s.This pathogen is prevalent primarily in temperate and subtropi-cal areas.Dogs,cats,horses,cattle,deer,and other animals are susceptible,and humans are also susceptible hosts.The main symptoms of Lyme disease in humans are erythema migrans,arthritis,and other neurological symptoms,and the common symptoms in infected animals include joint diseases,coat shedding,fever,laminitis,and lameness.Lyme disease is wide-spread,but diagnosis is difficult,and this disease is easily misdiagnosed and missed.Awareness of Lyme disease must be in-creased to avoid its toll on livestock and the pet industry.Therefore,this article reviews research progress in diagnosis and con-trol technology for animal Lyme disease and Borrelia burgdorferi,to provide a reference for accurate,rapid diagnosis and con-trol of Lyme disease.

The aim is to evaluate the effect of therapeutic communication on cervical cancer patients’ preoperative anxiety and hope level. The convenience sampling method was used to select the inpatients who will receive radical surgery for cervical cancer in the department of obstetrics and gynecology of the Second Affiliated Hospital of Xi’an Jiaotong University from November 2016 to November 2019 as the research object. 50 patients were grouped by the random number table method: 25 patients were in the intervention group, and 25 patients were in the control group. Patients in the intervention group were given therapeutic communication on the basis of routine nursing, and patients in the control group were given routine nursing. Both groups were investigated with the Self-Rating Anxiety Scale (SAS) and Herth Hope Index (HHI) on the first day of admission and the day before surgery. Before the intervention, there was no statistically significant difference between the two groups (P>0.05) . After the intervention, the anxiety level of the intervention group was lower than that of the control group (P<0.05), and the hope level was higher than that of the control group (P<0.05). It can be seen that therapeutic communication can alleviate preoperative anxiety of cervical cancer patients, improve their hope level, promote patient recovery, and ease tense medical relationship.

Objective This study investigated the impact of occupational mercury(Hg)exposure on human gene transcription and expression,and its potential biological mechanisms. Methods Differentially expressed genes related to Hg exposure were identified and validated using gene expression microarray analysis and extended validation.Hg-exposed cell models and PTEN low-expression models were established in vitro using 293T cells.PTEN gene expression was assessed using qRT-PCR,and Western blotting was used to measure PTEN,AKT,and PI3K protein levels.IL-6 expression was determined by ELISA. Results Combined findings from gene expression microarray analysis,bioinformatics,and population expansion validation indicated significant downregulation of the PTEN gene in the high-concentration Hg exposure group.In the Hg-exposed cell model(25 and 10 μmol/L),a significant decrease in PTEN expression was observed,accompanied by a significant increase in PI3K,AKT,and IL-6 expression.Similarly,a low-expression cell model demonstrated that PTEN gene knockdown led to a significant decrease in PTEN protein expression and a substantial increase in PI3K,AKT,and IL-6 levels. Conclusion This is the first study to report that Hg exposure downregulates the PTEN gene,activates the PI3K/AKT regulatory pathway,and increases the expression of inflammatory factors,ultimately resulting in kidney inflammation.

Objective To explore the correction of beam trajectories in the presence of fringe fields in magnetic resonance imaging-guided proton therapy and dose changes in the body before and after correction.Methods The open-source treatment planning software matRad was used to design plans for brain tumor,liver tumor,and prostate cancer cases,and simulation studies were conducted in the Monte Carlo simulation toolkit TOPAS to calculate proton dose distribution.A proton beam trajectory correction model suitable for three-dimensional magnetic fields was established,and a beam trajectory correction algorithm was developed.The deflection of the proton Bragg peak in the presence of fringe fields was analyzed.Furthermore,3 treatment plans were simulated and dose correction was carried out when the fringe field existed.Gamma analysis method is used to evaluate the correction effect;and the dose changes in the target area and organs-at-risk after correction were quantitatively analyzed.Results The perturbation of the magnetic field would cause lateral deflection of the proton beam trajectory,and the presence of fringe fields would significantly increase this effect,which increased with the increasing of beam energy.When the fringe field existed,the treatment plans for brain tumor,liver tumor,and prostate cancer were corrected.Under the 3%/3 mm criterion,the gamma passing rates for target area were 94.844%,92.054%,and 97.863%,respectively,and after correction,the total dose in the body was increased by 2.8%,2.5%,and 1.5%,respectively.The increased dose was mainly contributed by incident protons.Conclusion In magnetic resonance imaging-guided proton therapy,the effects of fringe fields should be considered.The increase in incident proton beam energy after correction will lead to an increase in the total dose in the body.Since the beam trajectory still has curvature,the dose changes differently in different organs-at-risk.

Objective To observe the effects of Tiaojing Cuyun Prescription on endometrial receptivity and AREG/EGFR/HIF-1α signaling pathway of embryo implantation dysfunction(EID)infertility mice;To explore its mechanism on improving endometrial receptivity.Methods On the first day of pregnancy,80 Kunming mice were randomly divided into control group,model group,progesterone group and Tiaojing Cuyun Prescription group,with 20 mice in each group.Progesterone group and Tiaojing Cuyun Prescription group were given corresponding drugs by gavage.On the fourth day of pregnancy,mifepristone solution was subcutaneously injected(2.3 mg/kg)to establish the EID infertility model except the normal group,while the normal group was subcutaneously injected with equal volume of propylene glycol.On the fifth day of pregnancy,10 mice were randomly selected from each group,and the endometrium morphology were observed by HE and Masson staining,the pinopodes were observed by scanning electron microscope,the content of AREG in endometrial tissue was detected by ELISA,and the expressions of EGFR,p-EGFR and HIF-1α were detected by Western blot and immunofluorescence,the rest of the mice were gavaged until the eighth day of pregnancy,and the average number of embryo implantation points was recorded.Results Compared with the normal group,the average number of embryo implantation points of model group significantly decreased(P<0.01),and the glandular development in the endometrium was poor,collagen fibers increased,and blood vessels and pinopodes decreased,the AREG content in endometrial tissue was significantly decreased(P<0.01),and the expressions of p-EGFR and HIF-1α protein were significantly decreased(P<0.05).Compared with the model group,the average number of embryo implantation points in Tiaojing Cuyun Prescription group significantly increased(P<0.05),the proliferation of collagen fibers in the endometrium was reduced,and the number of glands,blood vessels and pinopodes in stroma significantly increased,the development of pinopodes were full and uniform,the AREG content in endometrial tissue significantly increased(P<0.01),and the expressions of p-EGFR and HIF-1α protein significantly increased(P<0.05,P<0.01).The immunofluorescence results were consistent with Western blot.Conclusion Tiaojing Cuyun Prescription can promote the development of endometrial glands and blood vessels,enrich the number of pinopodes,improve the endometrial receptivity of EID infertility mice,facilitates embryo adhesion and implantation,and thus increase the pregnancy rate.Its mechanism may be related to activating AREG/EGFR/HIF-1α signaling pathway,improving endometrial perfusion,and restoring the relatively hypoxic environment of the endometrium.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective To investigate the distribution and antimicrobial resistance profiles of the common pathogens isolated from urine from 2015 to 2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods The bacterial strains were isolated from urine and identified routinely in 51 hospitals across China in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Antimicrobial susceptibility was determined by Kirby-Bauer method,automatic microbiological analysis system and E-test according to the unified protocol.Results A total of 261 893 nonduplicate strains were isolated from urine specimen from 2015 to 2021,of which gram-positive bacteria accounted for 23.8％(62 219/261 893),and gram-negative bacteria 76.2％(199 674/261 893).The most common species were E.coli(46.7％),E.faecium(10.4％),K.pneumoniae(9.8％),E.faecalis(8.7％),P.mirabilis(3.5％),P.aeruginosa(3.4％),SS.agalactiae(2.6％),and E.cloacae(2.1％).The strains were more frequently isolated from inpatients versus outpatients and emergency patients,from females versus males,and from adults versus children.The prevalence of ESBLs-producing strains in E.coli,K.pneumoniae and P.mirabilis was 53.2％,52.8％and 37.0％,respectively.The prevalence of carbapenem-resistant strains in E.coli,K.pneumoniae,P.aeruginosa and A.baumannii was 1.7％,18.5％,16.4％,and 40.3％,respectively.Lower than 10％of the E.faecalis isolates were resistant to ampicillin,nitrofurantoin,linezolid,vancomycin,teicoplanin and fosfomycin.More than 90％of the E.faecium isolates were ressitant to ampicillin,levofloxacin and erythromycin.The percentage of strains resistant to vancomycin,linezolid or teicoplanin was＜2％.The E.coli,K.pneumoniae,P.aeruginosa and A.baumannii strains isolated from ICU inpatients showed significantly higher resistance rates than the corresponding strains isolated from outpatients and non-ICU inpatients.Conclusions E.coli,Enterococcus and K.pneumoniae are the most common pathogens in urinary tract infection.The bacterial species and antimicrobial resistance of urinary isolates vary with different populations.More attention should be paid to antimicrobial resistance surveillance and reduce the irrational use of antimicrobial agents.