Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

[Purpose]To investigate the trends of incidence and age at onset of female breast can-cer in Jiangsu cancer registration areas from 2009 to 2019.[Methods]The continuous monitoring data of female breast cancer from 2009 to 2019 were collected from 16 cancer registries in Jiangsu Province.All datasets were checked and evaluated based on data quality control criteria and were included in the analysis.Crude rate(CR),age-standardized incidence rate by Chinese standard population(ASIRC),the average annual percentage change(AAPC),crude and adjusted mean age at onset were calculated.Incidence rates stratified by regions and age groups were calculated.Age-period-cohort model was used to analyze the changes of cancer incidence and age-adjusted mean ages.[Results]The incidence CR of female breast cancer was significantly increased from 24.39/105 in 2009 to 46.72/105 in 2019 with an AAPC in CR of 6.97％(95％CI:5.26％～8.70％)and AAPC in ASIRC of 4.67％(95％CI:3.00％～6.37％).The crude mean age and adjusted mean age at onset increased from 54.10 and 52.89 years old in 2009 to 56.23 and 53.36 years old in 2019,respectively.Crude mean age at onset increased significantly over time in all registry areas(β=0.21,P＜0.001),urban(β=0.1 8,P＜0.001)and rural(β=0.25,P＜0.001)areas,while adjusted mean age at onset remained stable in all registry areas(β=0.05,P=0.024)and rural areas(β=0.09,P=0.008).From 2009 to 2019,the incidence rate of female breast cancer of all age groups showed upward trends,with an average annual growth rate of 3.26％to 7.79％(all P＜0.05),and the incidence rate in rural areas increased faster than that in urban areas.The age composition of breast cancer onset and standardized age composition of onset in women over 60 years old showed an upward trend(all P＜0.05).[Conclusion]The incidence rate of female breast cancer in Jiangsu Province in-creased from 2009 to 2019,and the mean age of onset showed a backward trend.

Objective To prepare desalted Aconiti Lateralis Radix Praeparata(Danfupian)standard decoction for HPLC content determination and fingerprint analysis.Methods 15 batches of Danfupian standard decoction were prepared,and the transfer rate of monoester alkaloids was calculated by detecting their content.The transfer rates of monoester alkaloids of 15 batches of Danfupian standard decoction were calculated.The fingerprints of Danfupian standard decoction were established by HPLC,and the similarity values of fingerprints were calculated.Results According to the measurement of 15 batches of Danfupian standard decoction,the average transfer rate of monoester alkaloids was 78.03%with a rage from 56.93%to 94.06%.Besides,9 common peaks were determined and 3 peaks were calibrated.Moreover,the similarity was over 0.95 between fingerprints of standard decoction and the control fingerprint of the 15 batches of Danfupian standard decoction.Conclusion This study establishes an HPLC content determination and fingerprint analysis method of Danfupian standard decoction,which can provide reference for the quality control of the TCM granules of Danfupian.

Objective:To explore the value of conventional ultrasonography combined with cervical compression in the diagnosis of orbital venous malformation (OVM).Methods:A total of 43 patients with suspected OVM were admitted in sequentially from January 2019 to July 2022 in the Third Medical Center of PLA General Hospital. All patients were examined by ultrasonography combined with cervical compression and demonstrated by operation or digital subtraction angiography (DSA). The conventional ultrasound features of OVM were summarized, and the value of conventional ultrasonography combined with cervical compression in the diagnosis of OVM was discussed.Results:The features of the conventional ultrasound combined with cervical compression for diagnosis of OVM were as follows: the interior of lesion was mainly tubular structure, and the compression test was positive. After cervical compression, the lesion enlarged and the inner diameter of the internal tubular structure widened. Doppler flow imaging showed that the interior of lesion was mainly venous blood flow. Compared with the results of postoperative pathology or DSA, the sensitivity, specificity, accuracy and positive predictive value of ultrasonography combined with cervical compression were 0.952, 1.000, 95.3% and 100%, respectively. The results of Fisher exact diagnosis showed that there was no significant difference between ultrasonography and operation or DSA of OVM( P>0.05). Conclusions:Conventional ultrasound combined with cervical compression can be used as an effective method for the diagnosis of OVM.

Objective:To observe the influence of shared medical appointments on health-related quality of life and quality of sleep in patients after liver transplantation.Methods:By randomized controlled study, a total of 124 patients after liver transplantation were included from our hospital from January 2018 to January 2019, and according to the lottery method, all subjects were divided into the routine management group ( n=64) who received routine outpatient intervention and the shared medical management group ( n=60) who received shared medical appointments management. The health-related quality of life and quality of sleep were investigated and compared by post-liver transplant quality of life questionnaire (pLTQ) and Pittsburgh sleep quality index (PSQI) before intervention (the day of discharge) and after intervention (the end of the last shared outpatient service). Results:After intervention, the dimension scores of worry, economics, body function, emotional function, health service, complication and total score of pLTQ were improved in tow groups than before intervention [the routine management group: (41.90±7.61), (18.13±4.22), (22.22±5.10), (14.92±3.28), (20.39±4.87), (14.63±3.99), and (132.19±37.09) vs (32.25±5.55), (12.77±3.47), (17.58±4.72), (9.23±1.38), (15.17±4.81), (10.89±1.51) and (98.00±29.03) score, t=8.20, 7.85, 3.58, 12.79, 6.10, 7.01, 5.81, all P<0.001; shared medical management group: (46.12±7.92), (24.16±5.34), (25.55±5.42), (17.90±3.60), (24.81±5.12), (17.93±3.60) and (155.47±41.00) vs (32.57±5.69), (12.81±3.82), (17.00±4.70), (9.60±1.39), (15.39±4.84), (11.00±3.52) and (98.37±28.96) score, t=10.76, 13.39, 9.23, 16.66, 10.36, 10.66, 8.81, all P<0.001], and those in the shared medical management group were higher than those in routine management group ( t=3.03, 6.95, 3.53, 4.82, 4.93, 4.83, 3.32, all P<0.05). After intervention, the total score of PSQI scale were lower than before intervention in the routine management group [(10.48±2.14) vs (11.89±2.45) score, t=3.47, P=0.001], and the dimensions score of sleep quality, full-sleep time, sleep time, sleep efficiency, sleep disorders, daytime function, hypnotic and total score of PSQI were lower than before intervention in the shared medical management group [(1.41±0.32), (0.54±0.13), (1.17±0.26), (1.11±0.35), (1.21±0.27), (1.30±0.33), (1.08±0.21) and (8.05±1.75) vs (1.88±0.53), (0.86±0.37), (1.84±0.41), (2.05±0.56), (1.39±0.33), (1.47±0.43), (1.22±0.32) and (11.71±2.43) score, t=-5.88, -6.32, -10.69, -11.03, -3.27, -2.43, -3.65, -9.47, all P<0.05], and those in the shared medical management group were lower than those in routine management group ( t=-6.68, -6.39, -10.43, -10.97, -2.62, -2.12, -3.54, -6.90, all P<0.05). Conclusion:Shared medical appointments model can improve the health-related quality of life and quality of sleep in patients after liver transplantation, and improve the effectiveness of outpatient intervention.

Objective:To investigate the changes in gray matter volume (GMV) and abnormalities in structural covariant network (SCN) patterns in patients with chronic pontine infarction (PI).Methods:Patients with unilateral chronic PI (case group) with the first onset admitted to the First Affiliated Hospital of Zhengzhou University and Tianjin Medical University General Hospital from October 2014 to June 2021 were prospectively included. At the same time, healthy subjects matched with age, gender and education years (normal control group) were included. High-resolution three-dimensional T 1 structural MRI images and behavioral scores of the subjects were collected. The voxel-based morphometry and two-sample t test were used to explore the differences in GMV between the groups. Using GMV differential brain regions as seed points, SCN was constructed to explore the abnormality of structural covariant patterns in patients with PI. Spearman rank correlation analysis was used to analyze the correlation between GMV in differential brain regions and behavioral scores. Results:A total of 60 patients with PI were enrolled, including 33 left PI and 27 right PI, while 34 healthy controls were also enrolled. Compared with the normal control group, the GMV in bilateral posterior cerebellar lobe decreased significantly in the left PI group, and the GMV in left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe decreased significantly in the right PI group (Gaussian random field correction with voxel level P<0.001 and cluster level P<0.05, cluster voxel >20), and there was a significant correlation between GMV values in the left anterior and posterior cerebellar lobes and the right posterior cerebellar lobe and the motor function score ( P<0.05). In addition, compared with the normal control group, the right PI group had broader covariate brain regions and a significant increase in the number of structural connections between covariate brain regions (family-wise error correction with voxel level P<0.05, cluster voxel >20). Conclusions:The GMV in bilateral posterior cerebellar lobe decreases significantly in patients with chronic PI, and were secondary to broader covariate brain regions and structural connections. This may be the neural mechanism of impaired behavioral function in patients with PI.

Day surgery, as a new perioperative management model that is safe, efficient, and consumes a small amount of medical resources, has been widely recognized in recent years. With the requirements of China's new healthcare system reform and the promotion of national policies, there is a significant increase in hospitals carrying out day surgery, and they are about to enter a new stage of rapid development. Adequate preoperative evaluation of patients undergoing day surgery is an important link in improving perioperative care during day surgery. This article reviews the importance, location and personnel, methods, timing and content of day surgery preoperative evaluation, and current situation of day evaluation in China, in order to provide reference for clinical workers related to day surgery.

Objective:To explore the value of 5G robotic remote ultrasound in the diagnosis of plateau pulmonary edema(HAPE).Methods:A total of 27 patients who quickly entered Nagqu, Tibet at an altitude of 4 600 m-5 600 m from March to December 2021 and developed one of the clinical symptoms of HAPE were collected. All patients were examined by 5G remote robotic ultrasound and lung CT respectively. Kappa test was used to analyze the consistency of the two diagnostic results, and McNemar test was used to compare the difference in diagnostic results. The ROC curve was used to analyze the sensitivity and specificity of remote lung ultrasound scores in the diagnosis of HAPE.Results:Among the 27 patients, 16 showed thickening of pleural line, increasing of B line, lung consolidation, pleural effusion, etc. Meanwhile, 11 showed no abnormality. Additionally, 8 cases had diffuse pulmonary fluid in both lungs, and 8 cases had localized pulmonary fluid. ROC curve showed that the area under the curve of lung ultrasound score for the diagnosis of HAPE was 0.947 (95% CI=0.78-0.99, P<0.001). The sensitivity and specificity were 0.933 and 0.917, respectively. Lung CT diagnosis was positive in 15 cases. Lung CT showed thickening of lung texture, ground glass, small nodular shadow, fine reticulate shadow, etc. The diagnostic results of the two techniques were in good agreement (Kappa=0.924, P<0.001), and there was no significant difference between the two methods ( P>0.05). Conclusions:5G remote robotic ultrasound has high consistency with CT in the diagnosis of HAPE and is an alternative early diagnosis method for HAPE. It may have clinical application value in scattered medical resources and remote plateau areas.

Objective : To study the correlation between cerebral white matter structure with cognitive function and blood biochemical indexes in patients with end stage renal disease (ESRD) . Methods : The diffusion tensor imaging (DTI) data of 64 ESRD patients and 47 age and sex matched healthy people were collected. Tract⁃based spatial statistics (TBSS) and XTRACT analysis methods were used to compare the differences in diffusion parameters between the two groups. Pearson correlation analysis was used to detect the correlation between various diffusion parameters and blood biochemical indexes and cognitive related scales. Results : The values of FA in the ESRD group generally decreased (P < 0. 05) . The values of MD, AD and RD obviously increased (P < 0. 05) . The scores of mini mental state examination (MMSE) and Montreal cognitive assessment (MoCA) decreased (P < 0. 01), while that of trail making test A part (TMT⁃A) increased (P < 0. 05) . In the ESRD group, the values of FA in the right anterior thalamic radiation, optic radiation, acoustic radiation, and cingulum were negatively correlated with the levels of creatinine and urea nitrogen, and positively correlated with MoCA′s scores, and the values of MD and RD of these tracts were positively correlated with the concentration of urea. The values of FA in optic radiation, acoustic radiation and left temporal of cingulum were negatively correlated with the scores of TMT⁃A. The values of FA in the vertical occipital fasciculus, inferior fronto⁃occipital fasciculus, left middle longitudinal fasciculus and forceps major were negatively correlated with the concentration of creatinine. The values of FA in the left arcuate fasciculus were positively correlated with the MoCA′s score, the values of MD and RD of these tracts were positively correlated with the concentration of creatinine, and negatively correlated with the scores of MMSE and MoCA. The values of FA in the right superior longitudinal fasciculus, the right corticospinal tract and the right frontal aslant tract werepositively correlated with the MoCA′s score, the values of MD and RD of these tracts were positively correlated with the concentration of urea; the values of MD and RD in the left fornix were positively correlated with the level of creatinine and urea. Conclusion The structural integrity of white matter in ESRD patients is extensively damaged, which is significantly associated with a variety of cognitive impairments. Serum creatinine and urea nitrogen may be risk factors for the changes in white matter.

Objective To observe the safety and efficacy of regime that based on aqueous cytomegalovirus-DNA (CMV-DNA) load and IL-8 determination for therapeutic monitoring and local treatment cessation of cytomegalovirus retinitis (CMVR) patients after allogeneic hematopoietic stem cell transplantation (HSCT).Methods A prospective case series study.A total of 14 CMVR patients (22 eyes) after allogeneic HSCT diagnosed in Ophthalmology Department of Peking University People's Hospital between January 2016 and December 2018 were involved in this study.All patients were CMV-DNA seronegative at baseline and were treated with intravitreous injection of ganciclovir (IVG,3 mg in 0.05 ml) twice per week for 4 times in the induction stage and once a week in the maintenance stage.Aqueous humor sample was collected during the first time of IVG every week.CMV-DNA and the level of IL-8 were measured by real time quantitative PCR and ELISA,respectively.During follow-up,negative CMV-DNA (＜ 103/ml) or level of IL-8 ＜ 30 pg/ml in aqueous sample was set as local treatment cessation.Then patients were followed every 2 weeks for at least 6 months.BCVA,intraocular pressure and fundus examination were taken for each visit.The BCVA examination was performed using the international standard visual acuity chart,which was converted into logMAR visual acuity.BCVA and intraocular pressure at the baseline and the last follow-up were compared by the Student t matching test.Results Of the 14 CMVR patients (22 eyes) after allogeneic HSCT,8 patients (16 eyes) were bilateral,6 patients (6 eyes) were unilateral.At the baseline,the mean logMAR BCVA was 0.814 ± 0.563,the intraocular pressure was 17.2 ± 7.8 mmHg (1 mmHg=0.133 kPa),the mean aqueous CMV-DNA load was (3.43 ± 4.96)× 105/ml,the mean level of IL-8 was 518 ± 541 pg/ml.At cessation of local treatment,the median number of intravitreal injections was 5 times.Nine eyes showed negative CMV-DNA in aqueous humor,of which,7 eyes showed negative IL-8 in aqueous.CMV-DNA could still be detected in 13 eyes,while IL-8 was negative.Only one eye's retinal lesion was completely quiet.Six months after local treatment cessation,the mean logMAR B CVA was 0.812 ± 0.691,the intraocular pressure was 14.8± 5.4 mmHg;which was not significantly different from baseline (t=-0.107,1.517;P=0.916,0.137).Recurrence of CMVR happened in only 1 eye because of systemic EB virus infection.Retinal lesions progressively improved and became completely quiet in all the remaining 20 eyes.In 22 eyes,iatrogenic vitreous hemorrhage occurred due to low platelet count during treatment (＜ 30 × 109/ml) in 4 eyes.When the treatment was terminated for 6 months,the fundus of hematoma absorption was clearly visible.At the time of CMVR diagnosis,there were 2 eyes (9％) with posterior subcapsular opacity,which may be caused by systemic glucocorticoid therapy after allogeneic HSCT.Conclusion Aqueous CMV-DNA load and level of IL-8 could be used as quantitative variables for monitoring the therapeutic effect and determining time for local treatment cessation for CMVR after HSCT safely and efficiently.