To construct monoclonal antibodies against Haemonchus contortus native H11 protein.In this study,five 4-6 weeks female BALB/c mice were immunized with native H11 protein extrac-ted from adult worms by Concanavalin A lectin.Spleen cells were isolated and fused with SP2/0 cells after 3 times of immunization.Two hybridoma cell lines,named A1E3 and A10E1,which could stably secrete monoclonal antibodies against H11 protein were obtained.The subtype identi-fication and immunological analysis showed that the heavy chain of the two monoclonal antibodies belonged to IgG1 and the light chain was κ type,and both monoclonal antibodies recognized the natural antigen H11.Immunohistochemical localization and larval developmental inhibition test in vitro showed that the mAb A1E3 could be localized to the intestinal microvilli of the adult worm,and that the antibody can inhibit the growth of the fourth-stage larvae.The successful production of two monoclonal antibodies not only lays the foundation for the study of protective antigenic epitopes of the H11 protein and the development of epitope vaccines,but also provides a potential application of the monoclonal antibody for the treatment of haemonchusis in animals.

OBJECTIVE: To investigate the effect of vitamin D binding protein (DBP) gene polymorphism on susceptibility and prognosis of severe acute pancreatitis (SAP). METHODS: A prospective study was conducted. Eighty-three patients with SAP who were admitted to the department of general surgery of Tianjin Fifth Central Hospital from March 2018 to March 2021 were selected as the research objects, and 83 healthy people in the same period were selected as controls. Peripheral blood RNA was extracted and reverse transcribed into cDNA, and the genotype and allele frequency of DBP gene rs7041 locus were detected by fluorescence quantitative analyzer. Hardy-Weinberg equilibrium was used to test the genetic balance. On the day of admission, serum C-reactive protein (CRP) level was detected by scattering immunoturbidimetry, serum procalcitonin (PCT) level was detected by electrochemiluminescence, serum DBP level was detected by enzyme-linked immunosorbent assay (ELISA), and neutrophil to lymphocyte ratio (NLR) was calculated automatically by the instrument. The length of intensive care unit (ICU) stay, the length of hospital stay and prognosis during hospitalization of patients were statistically analyzed. Multivariate Logistic regression analysis was used to screen the influencing factors of SAP occurrence. RESULTS: The results of Hardy-Weinberg equilibrium test showed that the distribution of gene polymorphisms in the two groups of subjects conformed to the law of genetic equilibrium. The frequencies of TT genotype and T allele of DBP gene rs7041 locus in the patients of SAP group were significantly higher than those in the healthy control group [TT genotype: 34.94% (29/83) vs. 9.64% (8/83), T allele: 55.42% (92/166) vs. 38.55% (64/166), both P < 0.01], and the frequency of GT genotype was significantly lower than that in the healthy control group [40.96% (34/83) vs. 57.83% (48/83), P < 0.05]. There was no significant difference in the frequency of GG genotype between the healthy control group and SAP group [32.53% (27/83) vs. 24.10% (20/83), P > 0.05]. Further multivariate Logistic regression analysis showed that TT genotype [odds ratio (OR) = 2.831, 95% confidence interval (95%CI) was 1.582-5.067, P < 0.001] and T allele (OR = 2.533, 95%CI was 1.435-4.472, P < 0.001) of DBP gene rs7041 locus were independent risk factors for SAP in healthy people, while GT genotype was a protective factor for SAP (OR = 0.353, 95%CI was 0.143-0.868, P = 0.041). The levels of CRP, PCT, NLR and DBP in patients with TT genotype of DBP gene rs7041 locus were significantly higher than those in patients with GG/GT genotype on the day of admission in SAP group [CRP (mg/L): 43.25±13.25 vs. 31.86±12.83, PCT (μg/L): 1.53±0.24 vs. 1.21±0.20, NLR: 3.15±0.53 vs. 2.71±0.48, DBP (μg/L): 87.78±19.64 vs. 70.58±18.67, all P < 0.01]. The length of ICU stay in patients with TT genotype of DBP gene rs7041 locus in SAP group was significantly longer than that in patients with GG/GT genotype (days: 11.35±1.58 vs. 9.71±1.35, P < 0.01). The length of hospital stay of patients with TT genotype was longer than that of patients with GG/GT genotype (days: 23.41±3.64 vs. 23.17±3.57), and the in-hospital mortality was higher than that of patients with GG/GT genotype [34.48% (10/29) vs. 29.63% (16/54)], but the difference was not statistically significant (both P > 0.05). CONCLUSIONS The risk of SAP was significantly increased in patients with TT genotype of rs7041 locus of DBP gene, and the mechanism may be related to the increase of DBP expression. And carrying the TT genotype will prolong the ICU hospitalization time of SAP patients, but the effect on prognosis is not obvious.
Humans ; Polymorphism, Single Nucleotide ; Prospective Studies ; Vitamin D-Binding Protein/genetics* ; Acute Disease ; Pancreatitis/genetics* ; Genotype ; Prognosis

Objective:To investigate the techniques used in blood flow control of Kimura laparoscopic spleen-preserving pancreatectomy (LSPDP).Methods:Forty·five patients with benign or low-grade malignant pancreatic diseases undergoing LSPDP at Huzhou Central Hospital from May 2014 to Oct 2021 were analyzed retrospectively. Patients were divided into splenic vascular flow control group ( n=22) and routine management group ( n=23). Results:There was no significant difference in gender, age, BMI, accompanying symptoms, hypertension, diabetes, lesion size and pathological diagnosis between the two groups (all P>0.05). A higher overall spleen preservation rate (90.9% vs. 52.2%, χ2=8.213, P=0.004), lower incidence of morbidity with Clavien grade ≥ Ⅱ (22.7% vs. 73.9%, χ2=9.911, P=0.002) and shorter postoperative hospital stay [(9.6±4.5) d vs. (14.3±6.6) d, t=2.447, P=0.008] were achieved in the vascular flow control group compared with those in the routine group. Conclusion:Splenic vascular flow control techniques improve the success rate of spleen preservation in laparoscopic distal pancreatectomy, reduce the postoperative complications and shorten the postoperative hospital stay.

Objective:To investigate the therapeutic effect of exosomes divided from bone marrow mesenchymal stem cell (BMSC) on pancreatic cancer in vivo through regulation of tumor-associated macrophages (TAM) polarization.Methods:Ten male C57BL/6 mice weighing approximately 20 g, ages 4 weeks, were used for BMSC exosomes extraction and PKH26 labelling. Thirty female SPF BALB/c-nu/nu nude mice weighing approximately (18.56±0.85) g, ages 4-6 weeks, were adopted for pancreatic carcinoma models. The models were randomly divided into 3 groups with 10 in each: control group (injected with PBS through tail vein), portal vein treatment group (injected with exosomes in PBS through portal vein), tail vein treatment group (injected with exosomes in PBS through tail vein). After the models were executive 8 weeks later, the percentage of PKH26 positive-exosomes in pancreas tissue was quantified by flow cytometry technique. The volume of the primary pancreatic tumor, the tumor volume of inhibitory rate, the number of metastatic nodule, and the ascitic fluid were assess. Also, the weight of liver and the tumor weight were evaluated. The expression of M1 and M2 macrophage-activate biomarkers and the content of pancreatic cancer marker B7-H4 tumor carbohydrate antigen 199 in peripheral blood was detected. Moreover, the expression of Survivin and matrix metalloproteinase-9 (MMP-9) in pancreatic cancer tissue cells was also detected.Results:There was no significant difference for the tumor volume of inhibitory rate between portal vein treatment group (72.4±21.6)% and tail vein treatment group (70.1±20.7)% ( t=0.24, P=0.811). Compared with control group, the volume of the primary pancreatic tumor, the weight of liver, the tumor weight, the number and rate of liver metastatic nodule, the ascitic fluid, and the number of other metastatic nodule were totally lower in the 2 treatment groups with significant difference (all P<0.05). Compared with the control group, iNOS and CD68 in portal vein treatment group and tail vein treatment group were increased, while Arginase, CD206, B7-H4, tumor carbohydrate antigen 199, Survivin and matrix metalloprotein-9 were decreased, with statistically significant differences (all P<0.05). Conclusion:Exosomes derived from BMSC can inhibit the polarization of TAM to the M2 phenotype and induce their polarization to the M1 phenotype, thereby suppressing the proliferation, invasion and migration of pancreatic cancer.

Traumatic rib fractures are the most common injury in thoracic trauma. Previously，the patients with traumatic rib fractures were mostly treated non-surgically，of which 50%，especially those combined with flail chest presented chronic pain or chest wall deformities and over 30% had long-term disabilities，being unable to retain a full-time job. In the past two decades，thanks to the development of internal fixation material technology，the surgical treatment of rib fractures has achieved good outcomes. However，there are still some problems in clinical treatment，including inconsistency in surgical treatment and quality control in medical services. The current consensuses on the management of regional traumatic rib fractures published at home and abroad mainly focus on the guidance of the overall treatment decisions and plans，and relevant clinical guidelines abroad lacks progress in surgical treatment of rib fractures in recent years. Therefore，the Chinese Society of Traumatology affiliated to Chinese Medical Association and Chinese College of Trauma Surgeons affiliated to Chinese Medical Doctor Association，in conjunction with national multidisciplinary experts，formulate the Chinese Consensus for Surgical Treatment of Traumatic Rib Fractures（2021）following the principle of evidence-based medicine，scientific nature and practicality. This expert consensus puts forward some clear，applicable，and graded recommendations from aspects of preoperative imaging evaluation，surgical indications，timing of surgery，surgical methods，rib fracture sites for surgical fixation，internal fixation methods and material selections，treatment of combined injuries in rib fractures，in order to provide references for surgical treatment of traumatic rib fractures.

Objective:To identify which induced the symptoms/signs and laboratory abnormal findings occurred in patients with novel coronavirus pneumonia, by disease itself or by ribavirin and interferon-alpha treatments, through mining the adverse events (AEs) signals of the 2 antivirus agents.Methods:According to the symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia mentioned in the literature and "Diagnosis and Treatment scheme of Novel Coronavirus Pneumonia (trial version 5)" , AEs in this study were selected. Related data were collected from the U.S. FDA Adverse Events Reporting System (FARES) from Jan 1, 2004 to Dec 31, 2019, and the reporting odds ratio ( ROR) method was used for signals detection for the above-mentioned 2 drugs. Results:A total of 7 582 463 AEs related to drugs were reported in the FAERS database, of which 31 775 related to ribavirin and 2 345 related to interferon-alpha. The results showed that AEs related to ribavirin in respiratory, thoracic, and mediastinal disorders were nasal congestion, cough, laryngeal pain, pharyngeal oedema, productive cough, and dyspnoea; AEs related to interferon-alpha were laryngeal pain and haemoptysis. In other system organ class, AEs related to above 2 drugs were pyrexia, feeling cold, pyrexia, nausea, vomiting, diarrhoea, headache, arthralgia, myalgia, and rash. AEs of laboratory abnormal results related to ribavirin were white blood cell/platelet count decrease and aspartate/alanine aminotransferase increase; AEs related to interferon-alpha were white blood cell/platelet count decrease, aspartate/alanine aminotransferase increase, and lymphocyte count decrease.Conclusion:Some AEs induced by ribavirin and interferon-alpha were similar to symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia, which should be distinguished in the clinical practice.

Objective:To identify which induced the symptoms/signs and laboratory abnormal findings occurred in patients with novel coronavirus pneumonia, by disease itself or by ribavirin and interferon-alpha treatments, through mining the adverse events (AEs) signals of the 2 antivirus agents.Methods:According to the symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia mentioned in the literature and "Diagnosis and Treatment scheme of Novel Coronavirus Pneumonia (trial version 5)" , AEs in this study were selected. Related data were collected from the U.S. FDA Adverse Events Reporting System (FARES) from Jan 1, 2004 to Dec 31, 2019, and the reporting odds ratio ( ROR) method was used for signals detection for the above-mentioned 2 drugs. Results:A total of 7 582 463 AEs related to drugs were reported in the FAERS database, of which 31 775 related to ribavirin and 2 345 related to interferon-alpha. The results showed that AEs related to ribavirin in respiratory, thoracic, and mediastinal disorders were nasal congestion, cough, laryngeal pain, pharyngeal oedema, productive cough, and dyspnoea; AEs related to interferon-alpha were laryngeal pain and haemoptysis. In other system organ class, AEs related to above 2 drugs were pyrexia, feeling cold, pyrexia, nausea, vomiting, diarrhoea, headache, arthralgia, myalgia, and rash. AEs of laboratory abnormal results related to ribavirin were white blood cell/platelet count decrease and aspartate/alanine aminotransferase increase; AEs related to interferon-alpha were white blood cell/platelet count decrease, aspartate/alanine aminotransferase increase, and lymphocyte count decrease.Conclusion:Some AEs induced by ribavirin and interferon-alpha were similar to symptoms/signs and laboratory abnormal findings of novel coronavirus pneumonia, which should be distinguished in the clinical practice.

Objective To investigate effective approach to decrease portal venous hypertension and high perfusion of portal vein caused by small-for-size (SFS) liver graft transplantation with the aim of improving hepatocellular microcirculation.Methods Rat models with SFS liver graft (n =62) were well estab lished and divided into SFS group and trans-portal intrabepatic portosystemic shunt (TPIPSS) group.Hemodynamic parameters,histopathologically morphologic changes,postoperative complications,accumulated survival rate were recorded and analyzed.Venous filling time after liver reperfusion,hemodynamic parameters were evaluated using t test and Kruskal-Wallis test.Kaplan-Meier method was performed for survival analysis.Results Venous filling time after liver reperfusion was remarkably prolonged with the application of multihole cone-shaped tubes.Compared with SFS group,the filling time was 4-second longer in TPIPSS.At each endpoints of reperfusion within 90 mins,the portal vein pressures were lowered in the TPIPSS group than those of SFS group.Liver grafts were present with more regular structures in TPIPSS group,with no sign of hepatic sinusoid congestion or irregular clearance extension.In the aspect of postoperative complications,all the rat receivers showed ascites in the SFS group.Nevertheless,there was no ascites observed in TPIPSS rats,and 50％ rats (5/10) experienced clinical manifestations of hepatic encephalopathy.Persistent fever over 7 days was showed in 10％ rats (1/10) of SFS group and 40％ rats (4/10) of TPIPSS group,respectively.The mean survival was superior in TPIPSS group (37.2 ± 23.5) d than SFS group (17.7 ± 13.5) d,P ＜ 0.05.Conclusion TPIPSS could be a safe and feasible approach to improve portal venous hypertension caused by SFS liver graft and hepatocellular reperfusion.

Objective To study the effect of local mild hypothermia on serum brain natriuretic peptide (BNP) in patients with acute intracerebral hemorrhage.Methods Sixty patients with acute intracerebral hemorrhage were divided into local mild hypothermia group (30 cases) and routine therapy group (30 cases) by random digits table method.Routine therapy group was given conventional therapy,and local mild hypothermia group was given local mild hypothermia treatment besides conventional therapy.The neurologic impairment was evaluated according to American National Institute of Health Stroke Scale (NIHSS) scores on admission,and on the 3rd,7th and 14th day after treatment.Serum BNP levels in patients were determined dynamically on admission,and on the 3rd and 14th day after treatment.The effect was evaluated on the 14th day after treatment.Results There was no significant difference in serum BNP on admission between two groups (P ＞ 0.05).Serum BNP on the 3rd,14th day after treatment in local mild hypothermia group was lower than that in routine therapy group[(153.47 ± 32.01) ng/L vs.(187.45 ± 40.21)ng/L and (111.02 ± 38.27) ng/L vs.(139.71 ± 29.53) ng/L],and there was significant difference(P ＜ 0.01 or ＜ 0.05).There was no significant difference in NIHSS scores on admission and on the 3rd day after treatment between two groups (P ＞0.05).NIHSS scores on the 7th and 14th day after treatment in local mild hypothermia group was lower than that in routine therapy group [(13.84 ± 6.00) scores vs.(16.59 ± 4.62)scores and (9.23 ± 4.48) scores vs.(13.02 ± 6.76) scores],and there was significant difference (P ＜ 0.01).The total effective power in local mild hypothermia group was higher than that in routine therapy group[90.0％(27/30) vs.66.7％ (20/30)],and there was significant difference (P ＜ 0.05).Conclusions The local mild hypothermia therapy can not only significantly improve the defect of nerve function in patients with acute intracerebral hemorrhage but also reduce the serum BNP.It can improve the curative effect in patients with acute intracerebral hemorrhage.

ObjectiveTo demonstrate the efficacy and safety of Hangzhou tacrolimus capsule (Saishi Tac capsule,Hangzhou Zbongmei Huadong Pharmaceutical Co.Ltd,China) in Chinese liver transplant recipients.MethodsMulticenter,randomized open-labeled,prospective controlled clinical trial was performed in de novo Chinese liver transplant recipients.According to inclusive and exclusive criterion,83 liver recipients from 11transplant centers were enrolled.The recipients accepted Saishi Tac capsule,mycopheolate and steroid 48 h post-operation.The initial dose of Tac was 0.1-0.15 mg kg-1day-1and C0 was 8-12 ng/ml in the first 60 days,followed by 5-10 ng/ml until the terminal observation time poiut (12 weeks after transplantation).The efficacy and safety were estimated during the period.The primary efficacy endpoint of the study was the incidence of biopsy-confirmed acute rejection.Graft survival was the secondary endpoint.Safety was assessed by monitoring laboratory parameters and adverse events reported over the course of the study,such as infection,renal damage,hypertension,hyperlipema and diabetes mellitus and other adverse affairs.ResultsThe dose of Tac at 1st,2nd,4th and 8th week post-operation was (4.1±1.9),(4.5±2.1),(4.5±2.1),(4.4±1.8) and (4.1±2.1) mg,and correspondjng values to the C0 were (8.1±4.5),(8.9±4.5),(8.8±4.3),(8.8±4.1) and (8.0±2.8) ng/ml.During 12 weeks of follow-up,the incidence of biopsy-confirmed acute rejection was 4.8％ (4/83),and all of cases were reversed by implosive therapy.The survival rate of graft hver was 100％.The incidence of lung infection and diabetes mellitus was both 6.02％.ConclusionSaishi Tac capsule was safe and effective to Chinese liver transplant recipients.