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.

Background and Aims:Sleeve gastrectomy(SG)has become the most widely performed bariatric procedure worldwide,but postoperative gastroesophageal reflux disease(GERD)remains a major concern.This multicenter study aimed to identify independent risk factors associated with GERD after SG to guide preoperative assessment and intraoperative management.Methods:Clinical data of 672 patients who underwent SG between January 2020 and December 2022 in six bariatric centers and completed a 12-month follow-up were retrospectively analyzed.Demographic characteristics,esophagogastric junction(EGJ)integrity graded by the AFS system,operative parameters,and postoperative outcomes were compared between patients with and without GERD.Multivariate logistic regression was used to identify predictors of postoperative GERD.Results:The overall incidence of GERD after SG was 24.7%(166/672).Multivariate analysis revealed that a preoperative BMI>35 kg/m2(OR=1.68,P=0.033),EGJ integrity AFS grade>2(OR=2.90,P=0.006),and preoperative reflux symptoms(OR=2.44,P=0.030)were independent risk factors for GERD.A staple line more than 1 cm from the angle of His(OR=0.45,P<0.001)and a bougie size>36 Fr(OR=0.08,P=0.001)were protective factors.Conclusion:High BMI,impaired EGJ integrity,and preoperative reflux symptoms significantly increase the risk of GERD after SG,whereas adequate preservation of the His angle and appropriate bougie calibration may reduce it.Comprehensive preoperative EGJ assessment and standardized surgical techniques are essential for minimizing postoperative reflux.

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.

Background and Aims:Sleeve gastrectomy(SG)has become the most widely performed bariatric procedure worldwide,but postoperative gastroesophageal reflux disease(GERD)remains a major concern.This multicenter study aimed to identify independent risk factors associated with GERD after SG to guide preoperative assessment and intraoperative management.Methods:Clinical data of 672 patients who underwent SG between January 2020 and December 2022 in six bariatric centers and completed a 12-month follow-up were retrospectively analyzed.Demographic characteristics,esophagogastric junction(EGJ)integrity graded by the AFS system,operative parameters,and postoperative outcomes were compared between patients with and without GERD.Multivariate logistic regression was used to identify predictors of postoperative GERD.Results:The overall incidence of GERD after SG was 24.7%(166/672).Multivariate analysis revealed that a preoperative BMI>35 kg/m2(OR=1.68,P=0.033),EGJ integrity AFS grade>2(OR=2.90,P=0.006),and preoperative reflux symptoms(OR=2.44,P=0.030)were independent risk factors for GERD.A staple line more than 1 cm from the angle of His(OR=0.45,P<0.001)and a bougie size>36 Fr(OR=0.08,P=0.001)were protective factors.Conclusion:High BMI,impaired EGJ integrity,and preoperative reflux symptoms significantly increase the risk of GERD after SG,whereas adequate preservation of the His angle and appropriate bougie calibration may reduce it.Comprehensive preoperative EGJ assessment and standardized surgical techniques are essential for minimizing postoperative reflux.

Objective:To analyze the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021.Methods:Herpes zoster cases were collected from a surveillance system in Beijing City from December 2017 to April 2021. The cases included individuals under 20 years old from seven sentinel hospitals located in two districts (Miyun District and Changping District). The basic information, the rash date of rash onset and the location and number of lesions were investigated at the first visit to the hospital, and the lesion swab samples were collected for laboratory testing. A telephone follow-up was conducted 21 days after the onset of the rash to investigate the degree of pain, duration of the rash and duration of pain. The individuals who still experienced neuralgia were further investigated for their pain condition at 90 days after the onset of the rash, to discover cases with postherpetic neuralgia. DNA was extracted from the rash fluid, and the ORF62 gene region was amplified and sequenced to obtain the viral sequence. The wild-type strain or chickenpox vaccine strain was identified by using sequence alignment, and the clinical characteristics of cases with different varicella vaccinations were compared.Results:A total of 78 herpes zoster cases under 20 years old were investigated during 2017-2021 in Beijing City, and 61 cases completed the follow-up survey. The age range of 61 cases was 1.83 to 20.54 years with a median age of 17.50 years. There were 36 males (59.02%) and 25 females (40.98%). Among them, there were 29 cases with the chickenpox vaccine immunization history (18 cases with one dose, 5 cases with two doses and 6 cases with unknown doses), 13 cases with no vaccination history and 19 cases with unknown vaccination history. Among the 78 cases, the herpetic fluid samples of 64 cases were positive for VZV, including 62 cases identified as wild-type strains and two cases as vaccine strains. The two vaccine strain cases were both 2-year-old girls who had received one dose of varicella vaccine and developed herpes zoster 3 months and 13 months after vaccination. Among the 29 cases with chickenpox vaccine immunization history, the majority had 10 to 49 lesions, accounting for 58.62% (17 cases). The trunk was the most common site of lesions, accounting for 44.83% (13 cases). About 51.72% (15 cases) reported "no or mild" pain intensity. The median ( Q1, Q3) scores for the worst pain, duration of pain and the time to crusting of lesions in the herpes zoster cases were 3 (1.5, 5) points, 10 (1.5, 12.5) days and 10 (6.5, 13) days, respectively. There was no statistically significant difference in the constituent ratio of the location of lesions, number of lesions and pain degree among the cases with vaccination history, without vaccination history and with unknown vaccination history ( P>0.05). There was also no statistically significant difference in the distribution of pain score, duration of lesions and duration of pain across the three groups ( P>0.05). Conclusion:Wild strains are the predominant pathogens in herpes zoster cases under 20 years old in Beijing City during 2017-2021. The varicella vaccination has no significant impact on the clinical manifestations of herpes zoster cases.

Objective:To analyze the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021.Methods:Herpes zoster cases were collected from a surveillance system in Beijing City from December 2017 to April 2021. The cases included individuals under 20 years old from seven sentinel hospitals located in two districts (Miyun District and Changping District). The basic information, the rash date of rash onset and the location and number of lesions were investigated at the first visit to the hospital, and the lesion swab samples were collected for laboratory testing. A telephone follow-up was conducted 21 days after the onset of the rash to investigate the degree of pain, duration of the rash and duration of pain. The individuals who still experienced neuralgia were further investigated for their pain condition at 90 days after the onset of the rash, to discover cases with postherpetic neuralgia. DNA was extracted from the rash fluid, and the ORF62 gene region was amplified and sequenced to obtain the viral sequence. The wild-type strain or chickenpox vaccine strain was identified by using sequence alignment, and the clinical characteristics of cases with different varicella vaccinations were compared.Results:A total of 78 herpes zoster cases under 20 years old were investigated during 2017-2021 in Beijing City, and 61 cases completed the follow-up survey. The age range of 61 cases was 1.83 to 20.54 years with a median age of 17.50 years. There were 36 males (59.02%) and 25 females (40.98%). Among them, there were 29 cases with the chickenpox vaccine immunization history (18 cases with one dose, 5 cases with two doses and 6 cases with unknown doses), 13 cases with no vaccination history and 19 cases with unknown vaccination history. Among the 78 cases, the herpetic fluid samples of 64 cases were positive for VZV, including 62 cases identified as wild-type strains and two cases as vaccine strains. The two vaccine strain cases were both 2-year-old girls who had received one dose of varicella vaccine and developed herpes zoster 3 months and 13 months after vaccination. Among the 29 cases with chickenpox vaccine immunization history, the majority had 10 to 49 lesions, accounting for 58.62% (17 cases). The trunk was the most common site of lesions, accounting for 44.83% (13 cases). About 51.72% (15 cases) reported "no or mild" pain intensity. The median ( Q1, Q3) scores for the worst pain, duration of pain and the time to crusting of lesions in the herpes zoster cases were 3 (1.5, 5) points, 10 (1.5, 12.5) days and 10 (6.5, 13) days, respectively. There was no statistically significant difference in the constituent ratio of the location of lesions, number of lesions and pain degree among the cases with vaccination history, without vaccination history and with unknown vaccination history ( P>0.05). There was also no statistically significant difference in the distribution of pain score, duration of lesions and duration of pain across the three groups ( P>0.05). Conclusion:Wild strains are the predominant pathogens in herpes zoster cases under 20 years old in Beijing City during 2017-2021. The varicella vaccination has no significant impact on the clinical manifestations of herpes zoster cases.

Objective To evaluate the predictive value of ultrasonography of the respiratory variability index of inferior vena cava internal diameter (IVC-RVI) in preoperative fluid responsiveness evaluation of hip fracture in geriatric patients.Methods A retrospective case-control study was used to analyze 39 cases of elderly hip fractures admitted to the Daping Hospital of Army Medical Center from August 2017 to February 2018,including 15 males and 24 females,aged 65-92 years [(80.9 ±7.9)years].The heart rate (HR),mean arterial pressure(MAP),cardiac output (CO),and cardiac stoke volume (SV)were recorded five minutes after the patient entered the operating room and within 5 minures after capacity load test.The inferior vena cave during expiration (IVCe) and inferior vena cave during inspiration (IVCi) were measured by ultrasound before and after volume load test and the difference in IVC-RVI was calculated.Volumetric positive group were the cases of SV change (△SV) ≥15％ (positive group,n =21),and volume negative group were the cases of △SV ＜ 15％ (negative group,n=18).The correlations between various ultrasonic parameters and SV were analyzed by comparing HR,MAP,CO,SV,SV,IVCe,IVCi and IVC-RVI before and after the capacity load test.The predictive value of IVC-RVI for preoperative volumetric reactivity in elderly patients with hip fractures was evaluated by using the receiver operative characteristic (ROC) curve and Pearson correlation analysis.Results Prior to the volumetric load test,no significant difference was observed between the two groups concerning HR,MAP,CO,SV,and IVCe (P ＞ 0.05),IVCi in the positive group was significantly smaller than that in the negative group (P ＜ 0.05),and IVC-RVI in the positive group was significantly larger than that in the negative group (P ＜ 0.05).After the volumetric load test,there was no statistically significant difference between the two groups in CO,IVCe,IVCi,and IVC-RVI (P ＞ 0.05),HR in the positive group was smaller than that in the negative group (P ＜ 0.05),and MAP and SV in the positive group were significantly larger than that in the negative group (P ＜ 0.05).Compared with that before capacity load test,MAP,CO,and SV in the positive group increased after the capacity load test (P ＜ 0.05),while the HR difference was not statistically significant (P ＞ 0.05).HR,MAP,CO,and SV showed no significant difference in the negative group before and after capacity test (P ＞ 0.05).IVC-RVI assessed the area under the curve (AUC) was 0.80 ± 0.08 for preoperative volumetric responsiveness in elderly hip fractures (P ＜ 0.05),with a critical point of 20.69％,sensitivity of 77.78％,and specificity of 76.19％.After Pearson correlation analysis,IVC-RVI was positively related to △SV (OR =0.367,P ＜ 0.05).Conclusion Ultrasonography of the respiratory variability of IVC can predict the preoperative fluid responsiveness of hip fracture in geriatric patients and guide for clinical rehydration.

TCM believes that physical tendons and bone are interdependent, keeping dynamic balance, and they influence each other pathologically. Physical tendons and bone balance is human spine physiological state. Physical tendons and bone imbalance is the important pathogenesis of spine and related diseases. The core of physical tendons and bone balance-regulating theory is the physique homology and syndrome differentiation and treatment. This article expounded from the above aspects in details.

Objective To investigated the effects of patient controlled intravenous analgesia and ultrasound guided patient?controlled paravertebral block on pulmonary function in patients with multiple fractured ribs? Methods From May 2015 to March 2016,60 patients with multiple rib fractures in Ningbo NO?6 Hospital with American Society of Anesthesiologists Ⅱ-Ⅲ underwent internal fixation, including 30 males and 30 females aged from 16 to 70 years ( mean age ( 54?6 ± 15?2 ) years ) . All patients were randomly divided into 2 groups (n=30 each):patient controlled intravenous analgesia(PCIA) group and patient controlled thoracic paravertebral block(PCPB) group.PCIA solution contained sufentanil 2 μg/kg in 100 ml of normal saline. The PCIA pump was set up to deliver a 2 ml bolus dose with 15 minutes lockout interval and background infusion at 2 ml/hour.In PCPB group,the patients received paravertebral injection of 0?2% ropivocaine 15 ml on the corresponding area of fractured side under ultrasound guidance at the end of operation,and then received PCPB.PCPB solution contained 0?75% ropivacaine 67 ml in 250 ml of normal saline,and the pump was set up to deliver a 5 ml bolus dose, with 15 minutes lock out interval and background infusion at 5 ml/hour? Both analgesia lasted to 72 hours after operation.Before the operation,at 30 minutes,60 minutes,1 day,2 days,3 days after analgesia,arterial blood samples were collected to test the levels of pH,PaO2 ,PaCO2 ,PaO2/FiO2 and PA?aDO2 . The pulmonary function was also examined before and 3 days after the operation through FEV1 and FEV1%? Results Compared with PCIA group at 30 minutes, 60 minutes, 1 day, 2 days, 3 days after analgesia, the level of PaO2 ( ( 85?1 ± 9?7 ) mmHg vs? ( 72?4 ± 12?3)mmHg,(90?3±12?4)mmHg vs? (73?5±7?8)mmHg,(94?2±8?2)mmHg vs? (86?1±5?6)mmHg, (98?5±7?0)mmHg vs? (88?8±7?5)mmHg,(99?6±9?8)mmHg vs? (91?3±8?6)mmHg,P<0?05)) and PaO2/FiO2 were significantly increased ( ( 405?1 ± 46?0 ) mmHg vs? ( 340?1 ± 58?9 ) mmHg, ( 430?5 ± 59?1) mmHg vs? ( 344?0 ± 65?4 ) mmHg, ( 448?3 ± 39?1 ) mmHg vs? ( 410?1 ± 26?7 ) mmHg, ( 460?1 ± 33?5)mmHg vs? (423?2±36?5)mmHg,(465?1±28?2)mmHg vs? (435?1±40?8)mmHg,P<0?05)), the level of PA?aDO2 was decreased ((22?9±4?6)mmHg vs? (36?6±5?1)mmHg,(17?7±4?7)mmHg vs?(34?5±2?9)mmHg,(13?8±4?1)mmHg vs? (21?9±3?2)mmHg,(13?5±4?6)mmHg vs? (19?2± 3?8)mmHg,(12?4±2?0)mmHg vs? (17?7±2?4)mmHg,P<0?05)), and FEV1,FEV1% were significantly increased at 3 days after operation in PCPB group ((2?9±0?4)mmHg vs? (2?2±0?5)mmHg, (78?1± 4?7) mmHg vs? ( 64?8 ± 4?3 ) mmHg; P<0?01 ) ) . Conclusion Ultrasound guided patient?controlled paravertebral block improves the arterial oxygenation function and accelerates the recovery of pulmonary function in patients with multiple?fractured ribs after internal fixation operation.

Objective To investigated the effects of patient controlled intravenous analgesia and ultrasound guided patient?controlled paravertebral block on pulmonary function in patients with multiple fractured ribs? Methods From May 2015 to March 2016,60 patients with multiple rib fractures in Ningbo NO?6 Hospital with American Society of Anesthesiologists Ⅱ-Ⅲ underwent internal fixation, including 30 males and 30 females aged from 16 to 70 years ( mean age ( 54?6 ± 15?2 ) years ) . All patients were randomly divided into 2 groups (n=30 each):patient controlled intravenous analgesia(PCIA) group and patient controlled thoracic paravertebral block(PCPB) group.PCIA solution contained sufentanil 2 μg/kg in 100 ml of normal saline. The PCIA pump was set up to deliver a 2 ml bolus dose with 15 minutes lockout interval and background infusion at 2 ml/hour.In PCPB group,the patients received paravertebral injection of 0?2% ropivocaine 15 ml on the corresponding area of fractured side under ultrasound guidance at the end of operation,and then received PCPB.PCPB solution contained 0?75% ropivacaine 67 ml in 250 ml of normal saline,and the pump was set up to deliver a 5 ml bolus dose, with 15 minutes lock out interval and background infusion at 5 ml/hour? Both analgesia lasted to 72 hours after operation.Before the operation,at 30 minutes,60 minutes,1 day,2 days,3 days after analgesia,arterial blood samples were collected to test the levels of pH,PaO2 ,PaCO2 ,PaO2/FiO2 and PA?aDO2 . The pulmonary function was also examined before and 3 days after the operation through FEV1 and FEV1%? Results Compared with PCIA group at 30 minutes, 60 minutes, 1 day, 2 days, 3 days after analgesia, the level of PaO2 ( ( 85?1 ± 9?7 ) mmHg vs? ( 72?4 ± 12?3)mmHg,(90?3±12?4)mmHg vs? (73?5±7?8)mmHg,(94?2±8?2)mmHg vs? (86?1±5?6)mmHg, (98?5±7?0)mmHg vs? (88?8±7?5)mmHg,(99?6±9?8)mmHg vs? (91?3±8?6)mmHg,P<0?05)) and PaO2/FiO2 were significantly increased ( ( 405?1 ± 46?0 ) mmHg vs? ( 340?1 ± 58?9 ) mmHg, ( 430?5 ± 59?1) mmHg vs? ( 344?0 ± 65?4 ) mmHg, ( 448?3 ± 39?1 ) mmHg vs? ( 410?1 ± 26?7 ) mmHg, ( 460?1 ± 33?5)mmHg vs? (423?2±36?5)mmHg,(465?1±28?2)mmHg vs? (435?1±40?8)mmHg,P<0?05)), the level of PA?aDO2 was decreased ((22?9±4?6)mmHg vs? (36?6±5?1)mmHg,(17?7±4?7)mmHg vs?(34?5±2?9)mmHg,(13?8±4?1)mmHg vs? (21?9±3?2)mmHg,(13?5±4?6)mmHg vs? (19?2± 3?8)mmHg,(12?4±2?0)mmHg vs? (17?7±2?4)mmHg,P<0?05)), and FEV1,FEV1% were significantly increased at 3 days after operation in PCPB group ((2?9±0?4)mmHg vs? (2?2±0?5)mmHg, (78?1± 4?7) mmHg vs? ( 64?8 ± 4?3 ) mmHg; P<0?01 ) ) . Conclusion Ultrasound guided patient?controlled paravertebral block improves the arterial oxygenation function and accelerates the recovery of pulmonary function in patients with multiple?fractured ribs after internal fixation operation.