Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Objective:To investigate the clinicopathological features, and differential diagnosis of verrucous hemangioma (VH).Methods:Twenty-eight VH cases diagnosed from 2005 to 2020 in Henan Provincial People′s Hospital, Zhengzhou, China were analyzed retrospectively. Immunohistochemical studies were used to detect diagnostic markers. The mutation status of PIK3CA (exons 9 and 20) was detected using fluorescence PCR.Results:There were 13 males and 15 females in 28 cases, with the male to female ratio of 1.0∶1.2. There were 25 patients under the age of 18 years. The age range was from 10 months to 56 years (mean, 9.7 years; median, 4.5 years). There were 17 cases occurred in the lower extremities, 7 in the upper extremities and 4 in the trunk. All 28 cases were irregular red patches on the skin, which grew slowly. Some of them were thickened with uneven surface, which was light pink or red-white. Skin lesions of the 7 cases ranged from dark red and reddish brown, with a rough and hard surface. Satellite foci were present. Microscopically, 28 cases had a wide range of pathological features. Dilated, malformed vessels were observed from dermal papilla to deep soft tissue. Among them, the dermal papillary layer was mainly composed of many proliferating and expanding thin-walled capillaries and cavernous blood vessels. Thin-walled small vessels were found in the dermal reticular layer and subcutaneous fascia layer, with no obvious endothelial cell proliferation, occasional papillary hyperplasia, and lobular distribution of the malformed vessels in the fascia layer mixed with the fibroadipose tissue. There was epidermal papillary hyperplasia with hyperkeratosis and parakeratosis, lengthening and mutual fusion of epithelial horns. Immunohistochemistry showed that CD31, CD34, ERG and WT-1 were diffusely and strongly positive. The expression of GLUT-1 was present in superficial dermal vascular endothelial cells, but undetectable in the deep layer. The PIK3CA tests of 13 cases showed that no somatic mutations were found in exons 9 and 20. Twenty-five patients were followed up for 5 months to 10 years. Seven patients underwent multiple surgical resections and plastic surgeries due to the large size, and 8 patients had recurrence.Conclusions:VH is a rare congenital vascular malformation and more commonly occurs in infants and children. It tends to appear in limbs, especially lower limbs and distal limbs. Its morphology and immunophenotype are characteristic and should be distinguished from other vascular malformations and the resolution phase of infant hemangiomas. In about one third of the cases, postoperative recurrence may occur and long-term follow-up is often required.

For most stage T1 renal tumors, it is still necessary to block the renal pedicle to achieve the purpose of resection.However, it is not conducive to the recovery of postoperative renal function.Therefore, laser plays an important role in partial nephrectomy in order to achieve zero ischemia, and the aim of this review is to explain the research progress of laser in partial nephrectomy.

Single-port and reduced-port laparoscopic radical gastrectomy as the innovative surgery for gastric cancer are gradually accepted nowadays,and more attentions are also paid to single-port and reduced-port laparoscopic radical gastrectomy due to its better cosmetic effect,less pain,lower incidence rate of surgical site infection and more advantages in enhanced recovery after surgery.However,in the early stage,the development of single-port and reduced-port laparoscopic radical gastrectomy were facing challenges and obstacles on account of limited surgical skills,the lack of special laparoscopic instruments,laparoscope and multi-port Trocar.In recent years,the dilemma and difficult situations were gradually resolved following by surgical techniques innovation,laparoscopic instruments and facilities improvement.It is believed that single-port and reduced-port laparoscopic radical gastrectomy will have a good prospect and breakthrough in the field of gastric cancer treatment in the future.

The incidence of stroke increases year by year.It seriously affects the quality of life in patients.The pathogenesis of stroke is related to a variety of factors,involving genetic polymorphisms,biochemical mechanisms,and inflammatory effect.Among them,vascular endothelial growth factor (VEGF) has become one of the hotspots of research on the pathogenesis of stroke in recent years.This article reviews the correlation between VEGF gene polymorphism and stroke.

Objective To evaluate how proprioception affect ankle stability through comparing angle position awareness and peroneus reaction time between chronic ankle lateral instability patients and healthy controls.Methods A total of 51 participants were recruited into an experimental group of 21 patients with chronic ankle lateral instability (17 males,aged 31.6±2.6) and a control group of 30 healthy counterparts (24 males,aged 34.2±2.3).All the participants were asked to reoccur passive ankle position under the angular velocity at 2 degree per second when they were resting with non-weight bearing in their recruited ankles.The muscle reaction time (MRT)of peroneus longus(PL) and peroneus brevis (PB) in all the recruited ankles was measured during sudden ankle inversion both with and without ankle protective brace wearing.Results The difference between angle recurrence and the target angle (ankle inversion 20° and 30°) was significantly higher (P＜0.05) in the experimental group compared to the control group.The average MRTs of PL and PB were also significantly longer (P＜0.05) in the experimental group than the control group,whether wearing ankle protective braces or not.However,within both groups,no significant differences of PL and PB's MRT were identified between brace wearing and no brace (P＞0.05).Conclusions In patients with chronic lateral ankle instability,the position awareness decreases and the reaction time of peroneus is prolonged.Ankle braces can provide mechanical protection to the injured joints,but cannot promote MRT significantly.

Objective To investigate the effect of running exercise on cartilage in rats with an unstable knee joint.Methods Twenty 8-week-old Sprague-Dawley rats had their left anterior cruciate ligament cut to model an unstable knee.They were randomly divided into a control group and an experimental group,10 rats in each group.The control group was given no intervention,while the experimental group accepted running exercise training on na animal treadmill at a velocity of 15 m/min for an hour every day.After 3 and 6 weeks of training,5 rats were sacrificed and cartilage from the medial condyle of the femur was sampled,decalcificated,embedded and sliced on the sagittal plane.After hematoxylin-eosin staining,toluidine blue staining and immunohistochemical staining,the cartilage thickness,Mankin' score,the content of matrix collagen and the proteoglycan content of the cartilage matrix were assessed,and the shape and structure of the unstable knee joints were observed under a transmission electron microscope.Results The cartilage thicknesses and Mankin's scores at 6 weeks were significantly different from those at 3 weeks in both groups.In the experimental group the average thickness of cartilage was 154 ± 13 μm at 3 weeks and 131 ± 15 μm at 6 weeks.The corresponding Mankin's scores were 9.93 ± 1.36 and 11.23 ± 1.57,respectively.Both were significantly different from the control group averages at the same time points.There was also a significant difference in the positive rate of toluidine blue and collagen type Ⅱ staining between the experimental group and the control group at both time points,and in the experimental group between 3 and 6 weeks of training.After 3 weeks of training,fewer chondrocytes were observed under the transmission electron microscope in the experimental group,and fissures were seen on the surface of the cartilages.However,3 weeks later,quite a few ruptures and a lot of necrotic cells could be seen.Conclusions Running exercise can damage the cartilage of unstable knee joints and speed up the development of osteoarthritis.Even moderate exercise could aggravate damage to unstable joints and the cartilage matrix,and accelerate chondrocyte degeneration.

OBJECTIVE:To establish a method for the determination of 4 flavonoids constituents in Yinqiao capsules. METH-ODS:HPLC method was adopted. The Hypersil ODS C18 column was used with the mobile phase A of methanol-water-acetic acid (10∶88∶2,V/V/V)and B of methanol-water-acetic acid(88∶10∶2,V/V/V)in gradient elution at the flow rate of 1.0 ml/min;the de-tection wavelength was 327 nm,the column temperature was 25 ℃,and the volume was 10 μl. RESULTS:There was a good linear relationship between the amount of quercetin and peak area in the range of 0.050 9-1.018 0 μg(r=0.999 8),kaempferide in the range of 0.050 2-1.004 0 μg(r=0.999 5),isorhamnetin in the range of 0.051 0-1.020 0 μg(r=0.999 4)and rutin in the range of 0.050 4-1.007 0 μg(r=0.999 8). RSDs of precision,stability and repeatability tests were <2%. The average recoveries were 100.09%(RSD=0.93%,n=9),99.83%(RSD=0.75%,n=9),100.51%(RSD=1.17%,n=9) and 101.19%(RSD=1.08%,n=9). CONCLUSIONS:The method is amount specific,stable and reproducible and can be used for the quality control of Yinqiao capsules.

BACKGROUNDThe mutation frequencies of three common deafness genes (MT-RNR1 m.1555A>G, GJB2, and SLC26A4) among patients with nonsyndromic sensorineural hearing loss (NSHL) were different in previous studies. Inconsistent selection criteria for recruiting patients could have led to differences in estimating the frequencies of genetic mutations thus resulting in different mutation frequencies among these studies. The aim of this study was to reveal the differences in the mutation spectrums of the three common genes between familial and sporadic Chinese Han patients.

METHODSTotally, 301 familial probands and 703 sporadic patients with NSHL were enrolled in this study. Three genes, MT-RNR1 m.1555A>G, GJB2, and SLC26A4, were screened for mutation in our study cohort. A χ(2) test was performed to compare the mutation frequencies between the two groups.

RESULTSThe study showed that the disease-causing mutation frequencies of MT-RNR1 m.1555A>G, GJB2, and SLC26A4 were 12.29%, 14.62%, and 18.27% in familial probands and 3.56%, 18.63%, and 18.92% in sporadic patients, respectively. The mutation frequency of MT-RNR1 m.1555A>G in familial probands was significantly higher than in sporadic patients (χ(2) test, P = 0.000), while there were no significant differences in the mutation frequencies of GJB2 and SLC26A4 between the familial and sporadic groups (χ(2) test, P > 0.05).

CONCLUSIONSIt is necessary to reveal the differences in gene mutation frequencies between patients of different sources or characteristics by comparative studies in order to avoid selection bias. The mutations of GJB2, SLC26A4, and MT-RNR1 m.1555A>G are the most important etiological factors in Chinese Han patients, among which SLC26A4 might be the most frequent.

Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Connexin 26 ; Connexins ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genetic Testing ; Hearing Loss, Sensorineural ; genetics ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Middle Aged ; Mutation ; genetics ; Young Adult

Objective To discuss clinical,electroencephalogram(EEG) and PRRT2 gene mutation by reporting a febrile seizure (FS) with paroxysmal kinesigenic dyskinesia (PKD) family.Methods Detailed clinical data of the family were collected.The proband (Ⅳ1) and another 4 patients (Ⅲ1,Ⅲ4,Ⅳ2,Ⅳ3)were studied through clinical examinations.Clinical symptoms of Ⅳ2 were not typical,who was diagnosed as a suspected case.Mutation analysis of PRRT2 gene was screened by polymerase chain reaction (PCR) and DNA direct sequencing in 5 patients (Ⅳ1,Ⅲ1,Ⅲ4,Ⅳ2,Ⅳ3) and 4 unaffected family members (Ⅱ2,Ⅲ2,Ⅲ5,Ⅳ4).Results PKD patients had brief involuntary movements in the limbs or trunk induced by sudden voluntary movement when patients were in the stationary state since the teenagers.Two cases (Ⅲ,Ⅲ4) were accompanied by FS.Three cases(Ⅳ1,Ⅲ1 and Ⅲ4)had abnormal EEG records.The PRRT2 gene mutation (c.649dupC mutation) was identified in a healthy member (Ⅳ4) and 4 patients (Ⅳ1,Ⅲ1,Ⅲ4,Ⅳ3).Conclusions FS with PKD family has a PRRT2 gene mutation.The diagnosis is mainly based on family history,typical clinical manifestations and genetic test.This kind of disease may have pre-symptomatic patients.