To understand the immunogenicity of purified inactivated severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), concentration by ultrafiltration as well as molecular-sieve chromatography (MSC) were used for purification of inactivated SFTSVs. Inactivated viruses in purified samples were analyzed and identified by western blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the glycoprotein (GP) and nucleoprotein (NP) antigen titers of which were detected using a double-sandwich enzyme-linked immunosorbent assay (ELISA). Purified inactivated SFTSVs were enriched and observed by electron microscopy, and the total protein concentration detected using the bicinchoninic acid assay. Purified inactivated SFTSVs were applied to New Zealand rabbits via two immunization programs to evaluate immunogenicity and to compare the immune effect. After SFTSVs were inactivated and concentrated by ultrafiltration, MSC revealed two typical elution peaks. The sample of one peak was identified as inactivated virions, in which GP and NP were detected by SDS-PAGE, western blotting and ELISA. Main corponent of the other peak was NP. After concentration by ultrafiltration, purified inactivated SFTSVs with purity >90% and total protein concentration of 1. 1 mg/mL were obtained, and the typical electron microscopy of bunyavirus was observed. In the sera of animals immunized with purified inactivated SFTSVs, SFTSV-specific IgG antibody and neutralizing antibody were detected at high titers. However, antibody titers were affected by the immunization program. Effect of immunization on days 0, 14 and 28 was significantly better than that on days 0, 7 and 28. Our work revealed that cultivation of SFTSVs contained intact virus particles and large amounts of free NP. Using MSC, purified inactivated SFTSVs of high purity could be obtained. Purified inactivated SFTSVs induced high titers of neutralizing antibody and virus-specific IgG antibody showing satisfactory immunogenicity, which provides important clues for further study on a vaccine for the inactivated virus.
Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; immunology ; Bunyaviridae Infections ; immunology ; virology ; Humans ; Neutralization Tests ; Phlebovirus ; classification ; genetics ; immunology ; isolation & purification ; Rabbits

To prepare monoclonal antibodies (mAbs) against structural proteins of severe fever with thrombocytopenia syndrome bunyavirus (SFTSV), BALB/c mice were immunized using purified inactivated SFTSV virions as the antigens. Subsequently, hybridoma cell lines that secreted monoclonal antibodies against nucleoprotein (NP) and glycoproteins (GP) were obtained using a hybridoma technique. The antigen specificities of prepared mAbs were examined by indirect immunofluorescence and immunoprecipitation assays. Functional analyses were then performed,including the detection of IFA antibody titers,the levels of neutralizing activity and antibody affinities. After cell fusion and cloning,13 hybridoma cell lines secreted mAbs specifically against SFTSV-GP and 7 hybridoma cell lines secreted mAbs specifically against SFTSV-NP. Immunofluorescence and immunoprecipitation assays showed that the mAbs had high levels of antigen specificity. Among the 13 anti-SFTSV-GP mAbs,6 recognized Gn,whereas the others reacted with Gc. IFA titers of most anti-SFTSV-GP mAbs were between 1,280 and 20,480, and four anti-SFTSV-Gn mAbs showed neutralizing activity. Seven of the obtained anti-SFTSV-NP mAbs reacted specifically with NP,of which the IFA titers ranged from 5,120 to 20,480 with no observed neutralizing activity. Furthermore, two anti-SFTSV-GP mAbs, 1C8 and 1G8, showed high levels of affinity via a non-competitive ELISA. Our study lays the foundation for the development of further diagnostic assays and basic research into SFTSV.
Animals ; Antibodies, Monoclonal ; immunology ; Antibodies, Viral ; immunology ; Antibody Specificity ; Bunyaviridae Infections ; immunology ; virology ; Female ; Humans ; Hybridomas ; immunology ; Mice ; Mice, Inbred BALB C ; Phlebovirus ; immunology ; Viral Structural Proteins ; immunology

Objective To evaluate perfusion computed tomography in the assessment of portal vein pressure changes in an experimental dog model of liver cirrhosis and portal hypertension.Methods The canine model of cirrhosis and portal hypertension was induced by portal vein stenosis with combination of systemic thioacetamide(TAA) feeding in drinking water.All of the Beagles in control group and cirrhotic group underwent hepatic perfusion on a spiral CT scanner.The parameters of hepatic perfusion were calculated by the method of deconvolution.The portal vein pressure was measured by a laparotomy surgery.Results ① In control group, the portal vein pressure was ( 14.5 ± 2.2) cm H2O, while it was (23.1 ± 2.8) cm H2O in PHT group, there was significant difference in the portal vein pressure between the two groups (P＜0.05).② The blood flow(BF) was (112 ±14) ml·100 g-1·min-1 in controls, while ( 96 ± 11) ml·100 g-1·min-1 in PHT group; the blood volume ( BV ) in control group and PHT group was (10 ±3) ml·100 g-1 and (11 ± 5) ml· 100 g-1, respectively; the mean transit time( MTT) was (7.1 ± 2.0) s and (10.4 ± 3.5) s, respectively; the hepatic arterial fraction (HAF) was ( 24 ± 5) % and ( 37 ± 6)% , respectively; the hepatic arterial perfusion (HAP) was(27 ±6) ml·100 g-1·min-1 and (35 ±5) ml·100 g-1·min-1, respectively; the portal venous perfusion (PVP) was (85 ± 13) ml·100 g-1·min-1 and (61 ±11) ml·100 g-1·min-1, respectively.There was significant difference in all parameters between the two groups except the parameter BV(P ＜ 0.05).③ In PHT group, the PVP and BF were negatively correlated with the portal vein pressure, while positively correlated with MTT and HAF.Portal vein pressure was negatively correlated with PVP, the equation, Y = 36.624 -0.219X, was deduced with linear regression analysis, by which the portal vein pressure in PHT Beagles was ( 23.2 ± 2.4) cm H2O, which was correlated with the observed by laparotomy value (23.1 ± 2.8) cm H2O (r = 0.843, P ＜ 0.05).Conclusion CT perfusion is a new non-invasive and effective method for assessment of portal vein pressure.

OBJECTIVETo explore the association between occupational stress and other socio-psychological factors and work-related unintentional injury in off-shore oil production.

METHODSA cross-sectional survey was conducted among 561 Chinese off-shore oil workers. They were inquired with a self-administered questionnaire involving in socio-demographic characteristics, occupational stress, type A behavior personnel, social support and coping style, and work-related unintentional injury in the past year.

RESULTSSeventy-six (13.5%) workers experienced work-related unintentional injury for once time or more in the past year. A total 141 number of person-times were counted including 18 person-times being serious injury, 26 person-times moderate, and 97 person-times light. After adjustment for age, educational level, marital status, duration of off-shore work and job title, logistic regression indicated that perceived stress from "management problem and relationship with others" were significantly associated with work-related unintentional injuries in the past year (OR = 1.33, 95% CI: 1.05-1.68; P < 0.05). Workers lack of emotional support from friends seemed to less reporting on work-related unintentional injuries (OR = 0.37, 95% CI: 0.22-0.63).

CONCLUSIONThe results implied emphasizing on that the training and guidance as well as better cooperation among the workers might reduce work-related unintentional injuries in off-shore oil industry.

Accidents, Occupational ; prevention & control ; statistics & numerical data ; Adult ; Fuel Oils ; Humans ; Male ; Middle Aged ; Occupational Diseases ; complications ; Stress, Psychological ; complications ; Wounds and Injuries ; etiology ; prevention & control

Total mesorectal excision(TME)and complete mesoclic excision (CME)concepts make people aware of membrane integrity;the effect of endoscopic magnification and the hemostatic effect of ultrasonic scalpel,surgical field of view clearly,to further understand the structure of the membrane,which proposed the surgical anatomy of the membrane.This article describes the surgical membrane anatomy from the colorectal membrane of the embryonic development and membrane anatomical features that guide laparoscopic colorectal surgery.

Objective To explore the specific purification and enrichment method for the viral genome in clinical samples before high-throughput detection.Methods The negative chain RNA virus fever with thrombocytopenia syndrome (SFTSV) as was taken the simulate samples and the hybridization probes of the segment S,M and L were desiqned respectively,modified the probes with biotin in 5 ' ends to covalent bonded with streptavidin magnetic beads to purify the viral genome and compared the influence of specific purification on the effect of high-throughput detection.Results Specific enrichment and purification for viral genome before sequencing can significantly increase the proportion of valid data and whole genome coverage.The valid data proportions of the groups purified before or after whole transcriptome amplification (WTA) were improved significantly compared to group without purifying.The valid data proportion respectively were 5.57％,6.39％,2.71％,and the difference was significant((x2 =9799.8,P ＜0.001)).And the effect of the group purified after whole transcriptome amplification was better than group purified before whole transcriptome amplification(P ＜ 0.001).Conclusions Viral genome specific enrichment and purification method based on nucleic acid hybridization can effectively reduce the background values in the second generation sequencing,improve the sequencing efficiency and play the role of purification and enriching the nucleic acid in samples specifically before high-throughput detection.

Objective To elucidate the thermal stability and inactivation of severe fever with thrombocytopenia syndrome virus (SFTSV).Methods The stability of cell culture-derived SFTSV (SD4 strain) under different environmental temperatures,and the ability of heat,ultraviolet rays,diethyl ether,chloroform,acid,disinfectants treatments to inactivate SFTSV were evaluated.The infectious titers of treated viral samples were determined by titration assay based on a double N protein specific antibodies sandwich ELISA assay for SFTSV in Vero cells.An indirect immunofluorescence assay was performed to determine the replication dynamic of SFTSV in cells.Results SFTSV in culture medium was relatively stable at 4℃ without drastic loss of infectivity for at least 1 week,and the virus was sensitive to heat and could be inactivated in 30 min when incubated at 60℃.UV light irradiation with an intensity of 185 μW/cm2 efficiently inactivated SFTSV within 30 min.Exposures to optimized concentration of diethyl ether,chloroform,β-propionolactone,formaldehyde,Hypochlorous acid all destroyed SFTSV infectivity effectively.Acid conditions brought damage to the virus activity,but virus could not be completely inactivated at pH3.0.Conclusion The results provide quantitative evidence for the potential use of a variety of approaches for samples collection,inactivating SFTSV,and safety protection in scientific research and diseases control and prevention.

Objective:To investigate the effect of radiofrequency ablation in the treatment of severe tongue′s lymphatic vascular malformation, and evaluate the value and aesthetics of the operation.Methods:The 33 cases′ data of lingual lymphatic vascular malformation from February 2018 to February 2019 in Department of Hemangioma Surgery of Third Affiliated Hospital of Zhengzhou University were analyzed retrospectively. There were 18 males and 15 females, aged from 1 month to 50 years old, with a median age of 5 years. From unilateral lesions to diffuse and extensive lesions, 33 of them received radiofrequency ablation.24 cases were unilateral and 9 cases were bilateral. 18G radiofrequency ablation needle was selectedwithworking tip diameter of 1mm and heating 5 mm tip , and the power was adjusted to 25 W. Monitoring by color Doppler ultrasound, the working tip was inserted into the bottom of the lesion part which is about 1 mm from the deepest part. The ablation was performed for 5-20 s after being electrified. Then the electrode needle was withdrawn 1 mm in turn until it was 1 mm from the lesion surface, and respectively performed multi-point ablation withinterval of 5 mm.Results:One-stage radiofrequency ablation was performed in 24 patients with unilateral lesions and 9 patients with bilateral lesions. One-stage radiofrequency ablation was performed in one side of the lesions. After three to six months later, two-stage radiofrequency ablation was performed in most of the lesions. 33 cases were followed up from 6 months to 2 years.18 cases of unilateral lesions and 6 cases of bilateral lesions had no recurrence, and the tongue was symmetrical on both sides; 9 cases of full-thickness involvement of the tongue (6 cases on one side and 3 cases on both sides) had a small amount of lesions near the lingual artery, which were asymptomatic, nearly cured, without obvious scars, and satisfactory in appearance.Conclusions:Radiofrequency ablation is an important supplement for the treatment of lingual lymphatic vascular malformation, which makes up for the shortcomings of traditional surgery. Clinically, it can primarily achieve the treatment′s goal, with unobvious complication and aesthetical outcome.

Objective:To investigate the outcomes of therapeutic radiofrequency ablation (RFA) in unilateral intra-articular venous malformations (IAVMs) of the knee.Methods:The clinical data of 15 patients with intravascular venous malformation in the Third Affiliated Hospital of Zhengzhou University from February 2018 to May 2019 were retrospectively analyzed. There were 7 males and 8 females with an average age of 16 years (range, 5—25 years). All patients had unilateral knee involvement, ranging from localized lesions to diffuse extensive lesions. The radiofrequency ablation electrode needle with a diameter of 0.5 mm was selected and the power was adjusted to 25 W. Under the guidance of color ultrasound monitoring, the whole operation was carried out. The RFA electrode needle was inserted into the deepest site of the lesion and extracted by 1-mm graded movements until a point 1 mm from the surface of the lesion. Each ablation point was ablated for 10 to 30 seconds and multi-point ablation was done until the lesions were completely eliminated. Patients were followed up for 6 months to 1 year. Treatment outcomes were evaluated by functional activity, including extension, flexion, abduction, and adduction at the knee joint, and MRI findings, including intensity, shape, boundary, and degree of aggressiveness in the intra- and extraarticular cavities.Results:Included 15 patients; 12 had VMs in the knee joint cavity, and 3 had VMs involving both the intraarticular and extraarticular cavities (1 from the joint cavity to the calf, 1 from the joint cavity to the lower femur, and 1 from the intraarticular to the extraarticular cavity). All subjects underwent RFA, and the lesions were completely ablated and eradicated in a single operation. No recurrence was reported in the 6 months to 1 year follow-up. The knee joint remained symmetrical with the contralateral knee joint, without scarring, after the RFA.Conclusions:Radiofrequency ablation for the treatment of intra-articular venous malformation of the knee joint is a valuable minimally invasive surgical method with complete ablation, no complications, good prognosis and aesthetic appearance.

Objective:To investigate the effect of radiofrequency ablation in the treatment of severe tongue′s lymphatic vascular malformation, and evaluate the value and aesthetics of the operation.Methods:The 33 cases′ data of lingual lymphatic vascular malformation from February 2018 to February 2019 in Department of Hemangioma Surgery of Third Affiliated Hospital of Zhengzhou University were analyzed retrospectively. There were 18 males and 15 females, aged from 1 month to 50 years old, with a median age of 5 years. From unilateral lesions to diffuse and extensive lesions, 33 of them received radiofrequency ablation.24 cases were unilateral and 9 cases were bilateral. 18G radiofrequency ablation needle was selectedwithworking tip diameter of 1mm and heating 5 mm tip , and the power was adjusted to 25 W. Monitoring by color Doppler ultrasound, the working tip was inserted into the bottom of the lesion part which is about 1 mm from the deepest part. The ablation was performed for 5-20 s after being electrified. Then the electrode needle was withdrawn 1 mm in turn until it was 1 mm from the lesion surface, and respectively performed multi-point ablation withinterval of 5 mm.Results:One-stage radiofrequency ablation was performed in 24 patients with unilateral lesions and 9 patients with bilateral lesions. One-stage radiofrequency ablation was performed in one side of the lesions. After three to six months later, two-stage radiofrequency ablation was performed in most of the lesions. 33 cases were followed up from 6 months to 2 years.18 cases of unilateral lesions and 6 cases of bilateral lesions had no recurrence, and the tongue was symmetrical on both sides; 9 cases of full-thickness involvement of the tongue (6 cases on one side and 3 cases on both sides) had a small amount of lesions near the lingual artery, which were asymptomatic, nearly cured, without obvious scars, and satisfactory in appearance.Conclusions:Radiofrequency ablation is an important supplement for the treatment of lingual lymphatic vascular malformation, which makes up for the shortcomings of traditional surgery. Clinically, it can primarily achieve the treatment′s goal, with unobvious complication and aesthetical outcome.