Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To explore the short-term clinical efficacy, safety and patients' prognostic influencing factors of CyberKnife for the treatment of brain metastases in non-small cell lung cancer (NSCLC).Methods:A retrospective case series study was conducted. The clinical data of 58 NSCLC patients who received CyberKnife treatment for brain metastases at Jilin Cancer Hospital from July 2020 to January 2022 were retrospectively analyzed. At 3 months after CyberKnife treatment for brain metastases, and the efficacy of radiotherapy was evaluated on the basis of changes of brain metastases detected by contrast-enhanced magnetic resonance imaging (MRI) of the head. Overall survival (OS) and local recurrence-free survival (LRRFS) were analyzed in 58 patients by using the Kaplan-Meier method; the efficacy of cumulative brain metastasis volume for determining the survival of CyberKnife-treated NSCLC patients with brain metastases was analyzed by using the receiver operating characteristic (ROC) curve with the survival status of patients during the follow-up period as the gold standard, and the optimal cut-off value of cumulative brain metastasis volume was obtained; the clinical factors affecting OS and LRRFS of CyberKnife-treated NSCLC patients with brain metastases were analyzed by univariate and multivariate Cox proportional hazards models, and the adverse reactions associated with CyberKnife treatment were evaluated.Results:Among the 58 patients, 26 (44.8%) were male and 32 (55.2%) were female, with a median age [ M ( Q1, Q3)] of 64 years old (56 years old, 70 years old); there were 1-7 brain metastatic lesions in each patient, and there were 98 brain metastatic lesions in the 58 patients. There were 2 deaths (3.4%) within 3 months after CyberKnife treatment. At 3 months after treatment, there were 3 cases (5.4%) in complete remission, 36 cases (64.3%) in partial remission, 13 cases (23.2%) in stable disease, and 4 cases (7.1%) in disease progression in the remaining 56 patients. ROC curve analysis showed that the area under the curve for determining the survival of CyberKnife-treated NSCLC patients with brain metastases based on the cumulative brain metastasis volume was 0.593 (95% CI: 0.423-0.763), and the optimal cut-off value of cumulative brain metastasis volume was 15 cm 3. Median follow-up time was 12.6 months (7.5 months, 17.9 months). The 6- and 12-month OS rates were 91.3% and 79.5%, respectively, and the 6- and 12-month LRRFS rates were 93.0% and 89.2%, respectively. Multivariate Cox regression analysis showed that the Karnofsky functional status score (>70 points vs. ≤70 points, HR= 0.103, 95% CI: 0.019-0.545, P = 0.007), control of extracranial tumor (controlled vs. uncontrolled, HR = 0.145, 95% CI: 0.049-0.429, P < 0.001), cumulative brain metastasis volume (≤15 cm 3vs. >15 cm 3, HR = 0.105, 95% CI: 0.028-0.399, P = 0.001) were independent influencing factors for poor OS, and the control of extracranial tumor (controlled vs. uncontrolled, HR = 0.062, 95% CI: 0.006-0.616, P = 0.018), cumulative brain metastasis volume (≤15 cm 3vs. >15 cm 3, HR = 0.440, 95% CI: 0.007-0.292, P = 0.001), and target area total bioequivalent dose (BED) (≤60 Gy vs. >60 Gy, HR = 5.299, 95% CI: 1.020-27.530, P = 0.047) were independent influencing factors for poor LRRFS. Only grade 1-2 headache [53.5% (31/58)], nausea and vomiting [36.2% (21/58)] and other adverse reactions occurred after treatment, and no ≥grade 3 adverse reactions occurred. Conclusions:CyberKnife treatment for NSCLC brain metastases has high local control rate and short-term survival rate with mild adverse effects. Karnofsky functional status score, control of extracranial tumor and cumulative brain metastasis volume may affect OS of CyberKnife-treated NSCLC patients with brain metastases, and the control of extracranial tumor, cumulative brain metastasis volume and total BED may affect local recurrence.

Objective To understand and analyze the epidemiological and pathogenic characteristics of foodborne diseases in Changzhou in the past five years, so as to provide evidence for prevention and control measures. Methods All foodborne disease surveillance information reported by Changzhou foodborne disease sentinel hospital from 2016 to 2020 was collected through the ^“Foodborne Disease Surveillance and Reporting System^”. The distribution of reported cases and etiological results was described, and the possible factors affecting pathogen detection results were analyzed. Results A total of 14 931 cases of foodborne diseases were reported, mainly distributed in summer and autumn, and peaked in July and August. The age group of reported cases was 25-34 years old, and the occupations were mainly workers, and farmers and migrant workers, accounting for 23.2% and 36.9%, respectively. Among the reported cases of 3 120 anal swabs were collected and tested for etiology , of which 291 specimens were positive, with a positive detection rate of 9.32%. Among the 291 positive results of 118 cases of norovirus were detected, with a detection rate of 6.03% . 66 cases of salmonella were detected, with a detection rate of 2.12% . 25 cases of vibrio parahaemolyticus were detected, with a detection rate of 0.80% . 7 cases of diarrhea-causing Escherichia coli was detected, with a detection rate of 0.22% ; and 5 cases of shigella was detected, with a detection rate of 0.16% . Single factor analysis of the results of pathogen detection showed that there were no statistical differences in the distribution of positive pathogen detection rate between males and females, and between different suspicious exposed foods (all P>0.05). There were statistical differences in different age groups, occupations, seasons and eating places (all P<0.05). Multivariate Logistic regression analysis showed that age and season were correlated with the detection rates of pathogenic microorganisms (all P<0.05) , and age was negatively correlated with the positive rate of pathogenic microorganisms (OR=0.863). The detection rates in the first and fourth quarters were higher than those in the second and third quarters. Conclusion Summer and autumn are the seasons with high incidence of foodborne diseases in Changzhou. Norovirus is the main pathogen. Age and season are closely related to the detection results of pathogenic microorganisms. Comprehensive prevention and control measures should be implemented for key population in summer and autumn to reduce the occurrence and outbreak of foodborne diseases.

Background: This study aimed to explore relationships between job stress and psychological adaptation and how they related to interpersonal needs through mood states among female migrant manufacturing workers. Methods: A cross-sectional survey was conducted in 16 factories in Shenzhen, China. Sociodemographic, job stress, psychological adaptation and other psychological information of was collected. Structural equation modeling was performed to delineate the internal relationship between variables. Results: The hypothetical structural equation model exhibited acceptable model fit among female migrant manufacturing workers (χ2 = 11.635, df = 2, χ2/df = 5.82, p = 0.003, RMSEA = 0.090, CFI = 0.972, SRMR = 0.020). Job stress was directly associated with mood states and interpersonal needs; Psychological adaptation was directly associated with mood states and indirectly associated with interpersonal needs; Bootstrapping tests demonstrated mediation effect of mood states in the way from psychological adaptation to interpersonal needs. Conclusion Female migrant manufacturing workers who suffered stress from job and the process of psychological adaptation may have worse mood states and workers with worse mood states are more likely to develop unmet interpersonal needs, a proximal factor of suicidal ideation.

Objective:To explore the common risk factors of intracerebral hemorrhage(ICH) in young people and to establish a predictive model of nomogram.Methods:The relevant data of young patients with ICH (≤45 years ) hospitalized in the Department of Neurosurgery of Dezhou people's Hospital from January 2014 to August 2021 were retrospectively studied, and the young group who underwent physical examination in the Physical Examination Center of Dezhou people's Hospital at the same time were randomly selected as the control group. Analyze the risk factors that may affect cerebral hemorrhage in young people, screen the risk factors with statistical differences through single factor analysis, screen the independent risk factors according to multi factor Logistic regression analysis, construct the risk nomogram model of cerebral hemorrhage in young people, and test the efficiency, goodness of fit and benefit of the constructed model through internal validation.Results:Compared with the control group, there were statistically significant differences in family history (χ 2=115.66, P<0.001), hypertension grade( Z=17.67, P<0.001), smoking history (χ 2=33.91, P<0.001), drinking grade ( Z=4.84, P<0.001), body mass index (BMI) ( t=11.76, P<0.001), low density lipoprotein ( t=4.78, P<0.001), high density lipoprotein cholesterol ( t=5.83, P<0.001),blood glucose ( Z=5.68, P<0.001) and homocysteine ( Z=2.22, P<0.001) in the case group. Binary Logistic regression analysis showed that hypertension grade ( OR=3.457, 95%CI: 2.809-4.254, P<0.001), family history ( OR=2.871, 95%CI:1.868-4.413, P<0.001), BMI ( OR=1.093, 95%CI:1.040-1.148, P<0.001), high density lipoprotein cholesterol ( OR=0.230, 95%CI:0.111-0.480, P<0.001), blood glucose ( OR=3.457, 95%CI:2.809-4.254, P<0.001), homocysteine (O R=3.457, 95%CI:2.809-4.254, P<0.001) was an independent risk factor for intracerebral hemorrhage in young adults. The nomogram prediction model showed that BMI was 96 points, hypertension grade was 100 points, family history was 30 points, high density lipoprotein cholesterol was 76 points, homocysteine was 48 points, blood glucose was 52 points,homocysteine was 48 points and blood glucose was 52 points, respectively. The consistency coefficient of the prediction model was 0.874. The nomogram dependent ROC curve AUC was 0.891, and the corresponding sensitivity and specificity were 74.5% (263/353) and 89.7% (437/487), respectively, a nomogram model was established with good diagnostic efficiency. Conclusion:The nomogram model established in this study can predict the probability of intracerebral hemorrhage in high-risk population, and take intervention measures as early as possible to prevent the occurrence of intracerebral hemorrhage in young people.

Objective:To investigate the clinicopathological features of pulmonary epithelioid hemangioendothelioma (PEHE).Methods:Eighteen cases of PEHE were collected from August 2011 to December 2018 at the First Affiliated Hospital of Zhengzhou University. All cases were retrospectively studied by hematoxylin and eosin staining and immunohistochemistry (IHC). The clinicopathological features were reviewed; the status of CAMTA1 and TFE3 gene was analyzed and patients′ outcome was followed up.Results:Of the 18 cases, there were 11 males and 7 females with a male to female ratio of 1.6 to 1.0. The patients′ age ranged from 36 to 68 years (mean 52 years). Twelve cases (12/18) showed a single nodule and six cases (6/18) showed multiple bilateral nodules. Seven cases (7/18) involved other organs besides lung. Seventeen (17/18) patients presented with respiratory symptoms and one patient (1/18) presented with abdominal pain. Grossly, the tumors were greyish-white nodules with indistinct borders. Microscopically the tumor cells were epithelioid and arranged in strands and nests, and cytoplasmic vacuoles were commonly noted. The stroma was myxochondroid or hyaline. By IHC, the tumor cells were positive for CD31(18/18), CD34 (16/18), ERG (18/18) and Fli-1 (18/18); CKpan was focally positive in 5 cases (5/18). TFE3 was positive in 3 cases (3/18), and Ki-67 index ranged from 5% to 30%. FISH analysis showed seventeen cases (17/18) had CAMAT1 rearrangement, one case had TFE3 rearrangement displaying a split signal. Eight patients (8/18) had surgical excision, three patients (3/18) had surgery and chemotherapy, and seven patients (7/18) had chemotherapy only. Four patients (4/18) died of the disease.Conclusions:Patients with PEHE have non-specific symptoms, and correct diagnosis depends on pathologic biopsy and the exclusion of other tumors with epithelioid morphology. Some patients with PEHE have poor prognosis, particularly in those who have multiple nodules, peripheral invasion or metastasis.

Objective To explore the application of 3D printing technology in clinical apprenticeship teaching in spinal surgery. Methods 38 eight-year clinical medical students admitted to the Department of Spine Surgery were randomly divided into experimental group (19 patients) and control group (19 patients). Taking odontoid fracture as an example. The traditional method is used for teaching, and the experimental group is taught with traditional teaching method combined with 3D printing technology . After the apprenticeship, the students of the two groups were tested in theory and simulated operation to evaluate the teaching effect. SPSS 17.0 was used for statistical analysis. The measurement data was expressed as mean ± standard deviation, and the t-test was used to compare the measurement data between two groups. Results The results showed that the scores of immediate examination and one month after class in the control group were (82.23 ±7.52) and (72.36 ±6.39) points respectively. The scores of immediate examination and one month after the class in the experimental group were (85.58 ±6.32), (80.87 ±6.23). The scores of the two examinations in the experimental group were significantly higher than those in control group , and the differences were statistically significant (P<0.05). Conclusion The 3D printing technology can significantly improve the teaching effect in the clinical apprenticeship teaching of spine surgery, and it has a bright prospect for application.

Objective To establish a molecular method for the identification of different serotypes of group B streptococcus(GBS)based on TaqMan fluorescence probe technology,and to lay the foundation for the sub-sequent study of multiple fluorescent probe technology to detect different serotypes of GBS.Methods Primers and probes were designed according to the different serotypes of capsular polysaccharide(CPS).CPS se-quences were amplified by real-time fluorescence quantitative polymerase chain reaction.GBS classification methods of different serotypes were established.The results were compared with latex agglutination test and the method was evaluated from the aspects of sensitivity,specificity and detection of clinical isolates.Results The logarithmic concentration of DNA in the same serotype GBS was linearly correlated with the value of Ct. The detection limit of this method is 1 pg/μL,a probe could only detect the corresponding serotype GBS.The results of TaqMan fluorescence probe test of 10 strains were consistent with the results of latex agglutination test.Conclusion TaqMan fluorescence probe technique is a simple,rapid,highly sensitive and specific method for the detection of different GBS serotypes,and it is better than latex agglutination test for the classification of clinical isolates.

Objective To compare the pulp chamber temperature changes of Nd:YAP laser, Nd:YAG laser and semiconductor laser with the same power during dentin hypersensitivity treatment, and to evaluate the safety of these three laser treatments for dentin hypersensitivity. Methods 50 intact third molars were collected to prepare the dentin hypersensitivity model. The samples were randomly divided into Nd:YAP laser group (n=15), Nd:YAG laser group (n=15), semiconductor laser group (n=15), and blank control group (n=5). Each experimental group was divided into three subgroups (n=5) of 0.9 W, 1.4 W, and 1.8 W according to the laser power. The experiments were conducted with the corresponding laser parameters and thermocouple thermometer was used to record the temperature changes in the pulp chamber. The control group does not do any processing. After laser irradiation, one sample was randomly taken from each group and the morphology of dentin tubules was observed by scanning electron microscopy. Results When different power lasers were used to irradiate the samples, the temperatures of the pulp chamber in each group were increased. Among them, the temperature rise of the pulp chamber was smallest in the Nd:YAP laser group, followed by the semiconductor laser group, and the temperature rise was highest in the Nd:YAG laser group, but it was still lower than 5.5 ° C that could cause pulp necrosis. Scanning electron microscopy results showed that after irradiation with different power lasers, the diameters of most dentinal tubules in the Nd:YAG laser group and the semiconductor laser group were narrowed or even melted, and the effect was better than that of the Nd:YAP laser group. Conclusion The treatment using Nd:YAP laser, Nd:YAG laser and semiconductor laser for dentin hypersensitivity will increase the temperature of the pulp chamber. However, the temperature rise is less than 5.5℃and that will not cause irreversible damage to the pulp tissue. Nd:YAG laser and semiconductor laser have better dentinal tubule blocking effect, which is more suitable for laser dentin desensitization treatment than Nd:YAP laser.