Understanding the complexity of Tityus serrulatus venom: A focus on high molecular weight components

Tityus serrulatus scorpion is responsible for a significant number of envenomings in Brazil, ranging from mild to severe, and in some cases, leading to fatalities. While supportive care is the primary treatment modality, moderate and severe cases require antivenom administration despite potential limitations and adverse effects. The remarkable proliferation of T. serrulatus scorpions, attributed to their biology and asexual reproduction, contributes to a high incidence of envenomation. T. serrulatus scorpion venom predominantly consists of short proteins acting as neurotoxins (α and ß), that primarily target ion channels. Nevertheless, high molecular weight compounds, including metalloproteases, serine proteases, phospholipases, and hyaluronidases, are also present in the venom. These compounds play a crucial role in envenomation, influencing the severity of symptoms and the spread of venom. This review endeavors to comprehensively understand the T. serrulatus scorpion venom by elucidating the primary high molecular weight compounds and exploring their potential contributions to envenomation. Understanding these compounds' mechanisms of action can aid in developing more effective treatments and prevention strategies, ultimately mitigating the impact of scorpion envenomation on public health in Brazil.

Effect of cell migration-inducing hyaluronidase 1 on biological function of gallbladder cancer GBC-SD cells and its mechanism / 中华肝胆外科杂志

Objective:To investigate the effect of cell migration-inducing hyaluronidase 1 (CEMIP) on biological function of gallbladder cancer GBC-SD cells and its possible mechanism.Methods:The expression of CEMIP in biliary epithelial cell line HIBEC and gallbladder cancer cell line NOZ and GBC-SD was detected by Western blot. GBC-SD cells in logarithmic growth phase were divided into blank control group, negative control group (transfection with nonsense siRNA), siCEMIP-1 group (transfection with siCEMIP-1) and siCEMIP-2 group (transfection with siCEMIP-2). The expression of CEMIP and binding immunoglobulin protein (Bip) and calreticulin (CRT) in GBC-SD cells was detected by Western blot after culturing for 48h in blank control group, negative control group, siCEMIP-1 and siCEMIP-2 group. The relative survival rate was determined by CCK-8 assay. The wound healing rate and apoptotic rate was detected by wound healing assay and flow cytometry. The migration and invasion abilities were evaluated by Transwell chamber assay. Twelve 5-week-old BALB/c-nude mice were selected and divided into control group and experimental group (6 mice in each group). GBC-SD cells and GBC-SD cells with silenced CEMIP were subcutaneously injected into the right armpit (forelimb) of the two groups of mice, respectively. The volume and weight of transplanted tumor were compared 33 days later.Results:Compared with HIBEC cells, the relative protein level of CEMIP in gallbladder cancer GBC-SD cells [(0.750±0.034) vs. (0.120±0.002)] and NOZ cells [(0.690±0.013) vs. (0.120±0.002)] was significantly higher ( P<0.05). Compared with blank control group and negative control group, the relative protein level of CEMIP, Bip and CRT in siCEMIP-1 group and siCEMIP-2 group was significantly lower ( P<0.05). Compared with blank control group and negative control group, the relative survival rate and wound healing rate and number of migration cells and invading cells in siCEMIP-1 group and siCEMIP-2 group were also significantly lower ( P<0.05). While the apoptotic rate in siCEMIP-1 group and siCEMIP-2 group were higher than that in blank control group and negative control group ( P<0.05). Compared with control group, the average tumor volume [(543.6±114.7) vs. (801.5±256.3) mm 3] and tumor weight [(0.453±0.093) vs. (0.728±0.278) g ] of the experimental group was significantly decreased ( P<0.05). Conclusions:CEMIP was up-regulated in gallbladder cancer cell line GBC-SD and NOZ. Silencing CEMIP inhibited cell proliferation, wound healing rate, migration and invasion, and promoted apoptosis in gallbladder cancer GBC-SD cells, which may be related to the inhibition of endoplasmic reticulum chaperone Bip and CRT expression.

Effectiveness of FibroTouch combined with four hepatic fibrosis biomarkers for evaluation of the liver fibrosis degree among patients with chronic schistosomiasis-induced liver disorders / 中国血吸虫病防治杂志

Objective To evaluate the performance of FibroTouch in combination with four hepatic fibrosis biomarkers for assessment of the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders. Methods A total of 63 patients with chronic schistosomiasis-induced liver diseases admitted to The Third People’s Hospital of Kunshan City from January to March 2021 were enrolled as the observation group, while 50 healthy volunteers receiving health examinations in the hospital during the study period were randomly selected as the control group. The liver stiffness measurement (LSM) was determined using the FibroTouch technique, and the serum levels of four hepatic fibrosis biomarkers were detected using chemilumi-nescence immunoassay, including type IV collagen (IV-C), type III procollagen (PC-III), hyaluronidase (HA) and laminin (LN). The receiver operating characteristic (ROC) curves of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders were plotted and the area under the ROC curve (AUC) was estimated to examine the value of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis. Results There were 63 subjects in the observation group, including 28 men and 35 women, and the participants had a mean age of (65.34 ± 12.56) years and a mean body mass index (BMI) of (24.47 ± 11.05) kg/m2. There were 50 subjects in the control group, including 22 men and 28 women, and the participants had a mean age of (64.28 ± 13.10) years and a mean BMI of (25.12 ± 11.64) kg/m2. There were no significant differences between the observation and control groups in terms of gender ratio (χ2 = 0.002, P > 0.05), age (t = 0.437, P > 0.05) or BMI (t = 0.303, P > 0.05). The LSM [(8.65 ± 5.22) vs. (3.24 ± 1.10) kPa; t = 8.013, P < 0.05], IV-C [(51.80 ± 9.45) vs. (30.10 ± 10.34) ng/L; t = 11.506, P < 0.05], PC-III [(77.28 ± 17.22) vs. (48.62 ± 9.54) ng/L; t = 11.224, P < 0.05], HA [(39.55 ± 5.32) vs. (84.89 ± 10.34) ng/L; t = 30.158, P < 0.05] and LN [(99.47 ± 7.37) vs. (61.93 ± 9.80) ng/L; t = 22.496, P < 0.05] were significantly greater in the observation group than in the control group, and Spearman correlation analysis showed that the degree of liver fibrosis positively correlated with LSM (rs = 0.675, P < 0.01), IV-C (rs = 0.421, P < 0.01), PC-III (rs = 0.517, P < 0.01), HA (rs = 0.550, P < 0.01) and LN (rs = 0.539, P < 0.01) among patients with chronic schistosomiasis-induced liver diseases. ROC curve analysis revealed that the AUC of LSM for assessment of the hepatic fibrosis degree was 0.884 (P < 0.001), and the LSM cutoff, sensitivity and specificity were 11.75 kPa, 71.43% and 84.00% at the highest Youden index, respectively. In addition, the AUC of four hepatic fibrosis biomarkers for assessment of the hepatic fibrosis degree was 0.577 to 0.670, with 70.174 to 115.237 ng/L cutoff values, 17.46% to 68.25% sensitivity and 71.01% to 96.00% specificity. In addition, the sensitivity and specificity of LSM combined with four hepatic fibrosis biomarkers were 92.06% and 95.07% for assessment of the hepatic fibrosis degree among patients with chronic schistosomiasis-induced liver diseases. Conclusion FibroTouch in combination with detection of four hepatic fibrosis biomarkers has a high sensitivity and specificity for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver diseases, which deserves widespread clinical uses.

Sodium nitroprusside combined with hyaluronidase and urokinase in treatment of superficial epigastric arterial embolization in Sprague-Dawley rats / 中华医学美学美容杂志

Objective:To explore the effect of intravascular sodium nitroprusside (SNP) combined with hyaluronidase (HAase) plus urokinase (UK) in treating rat abdominal wall skin flap ischemia caused by HA induced artery embolism, and to compare the difference between thrombolysis with and without addition of sodium nitroprusside.Methods:Forty male Sprague-Dawley rats were injected with 10 μl of hyaluronic acid (HA) into the left inferior epigastric arteries, constructing the superficial epigastricl artery skin flap ischemic model in rats. The rats were randomly divided into four groups: a control group, and experimental groups A, B and C. Control and experimental groups A, B and C were treated with the following solutions 45 minutes after hyaluronic acid injection: physiological saline plus glucose was injected into the rats (control group); hyaluronidase (HAase) plus glucose injection was injected into the rats (experimental group A), hyaluronidase (HAase) plus urokinase (UK) was injected into the rats (experimental group B), hyaluronidase (HAase), urokinase (UK) plus sodium nitroprusside were injected into the rats (experimental group C). The changes of flaps were observed at 0 min, 3 days, 5 days and 7 days after operation. The difference of the area percentage of unperfused flap in the four groups was compared 7 days after operation. This study was carried out from July 2020 to March 2021 in the Medical Laboratory Animal Center of Weifang Medical University.Results:The unperfused area of flaps for the control group, experimental groups A, B and C were (100.00±0.00) %, (44.68±7.90)%, (34.01±8.77)% and (24.12±4.58)%, respectively. In the experimental group C, the scabby necrosis area was smaller than that of the experimental group A ( P<0.05); in the experimental group C, the scabby necrosis area was smaller than that of the experimental group B ( P<0.05); in the experimental group B, the scabby necrosis area was smaller than that of the experimental group A ( P<0.05). HE staining revealed that size and density of the embolus was significantly decreased after the addition of sodium nitroprusside. Conclusions:Sodium nitroprusside combined with hyaluronidase and urokinase can effectively improve the ischemia of the flap caused by HA induced artery embolism, increase the tissue perfusion, and reduce the necrotic area of the flap.

Errores innatos de la inmunidad en adultos / Inborn errors of immunity in adults

Introducción: La inmunodeficiencia común variable es un error innato de la inmunidad que tiene su pico de incidencia en la edad adulta. Se caracteriza por una susceptibilidad aumentada a padecer infecciones respiratorias, autoinmunidad y malignidad, secundario a un estado de hipogammaglobulinemia e inmunodisregulación, causado por mutaciones e interacciones genéticas parcialmente comprendidas. El diagnóstico es de exclusión, tiene una gran heterogeneidad clínica y comúnmente es diagnosticado de forma errónea. Objetivo: Describir un caso clínico de un paciente afectado por un error innato de la inmunidad. Caso clínico: Hombre de 35 años que se presenta a la consulta de Medicina Interna - Inmunología refiriendo un cuadro clínico de 3 años de evolución consistente en múltiples episodios de infecciones sino-pulmonares en los últimos meses, presentaba tos productiva, dificultad respiratoria y pérdida de peso no intencional de aproximadamente 8 kg. Conclusiones: La inmunodeficiencia común variable debe considerarse dentro de los diagnósticos diferenciales en todo paciente que presente alguna de sus manifestaciones clínicas, principalmente aquellas relacionadas con infecciones respiratorias a repetición, antecedente que el paciente puede presentar como relevante en sus consultas de primer nivel con medicina general o con especialistas. Su aproximación diagnóstica consiste en la solicitud de niveles séricos de inmunoglobulinas, prueba de laboratorio de fácil acceso para cualquier clínico independiente de su nivel de atención y su tratamiento se fundamenta en la administración periódica de inmunoglobulina humana exógena de forma endovenosa o subcutánea(AU)

Introduction: Common variable immunodeficiency is an inborn error of immunity that has its peak incidence in adulthood. It is characterized by an increased susceptibility to respiratory infections, autoimmunity and malignancy, secondary to a state of hypogammaglobulinemia and immunodysregulation, caused by mutations and partially understood genetic interactions. The diagnosis is one of exclusion, has great clinical heterogeneity and is commonly misinterpreted. Objective: To describe a clinical case of a patient affected by an inborn error of immunity. Methods: Retrospective description of a case report. Conclusions: Common variable immunodeficiency disorder should be considered within the differential diagnoses in every patient who presents any of its clinical manifestations, mainly those related to recurrent respiratory infections, an antecedent that the patient may present as relevant during the first-level consultations with general medicine physicians or with specialists. Its diagnostic approach consists in measuring serum immunoglobulin levels, an easily accessible laboratory test for any clinic physician regardless of their healthcare level, while its treatment is based on the periodic administration of exogenous human immunoglobulin intravenously or subcutaneously(AU)

Loxoscelismo cutáneo-visceral en una zona rural con desenlace favorable / Viscerocutaneous loxoscelism with a favorable outcome in a rural area

En el Perú los accidentes por mordedura de araña representan un problema de salud colectiva; de estos, el producido por la araña Loxosceles ha sido causa de numerosas muertes sobre todo en la costa. Esta tiende a ocasionar, ya sea una lesión cutánea o un cuadro sistémico, que puede llevar a la muerte del individuo si no es tratado a tiempo. Hasta la fecha no se cuenta con un protocolo de diagnóstico, predicción ni manejo a nivel internacional, por lo que se utilizan opciones terapéuticas sin respaldo de evidencia. Sin embargo, el manejo de soporte oportuno y adecuado es crucial para los cuadros severos. Se presenta el caso inusual de un loxoscelismo cutáneo-visceral o también llamado sistémico ocurrido en una zona rural. Hubo un compromiso renal severo que requirió hemodiálisis con un desenlace favorable a pesar del no uso de suero antiloxoscélico, lo que evidencia la importancia del manejo oportuno con las medidas de soporte adecuadas(AU)

Accidents caused by spider bites are a public health problem in Peru. Of these, those related to Loxosceles spider bites have been the cause of numerous deaths, mainly on the coast. These bites generally result in a cutaneous lesion or systemic involvement, which may threaten the person's life if not treated timely. An international protocol is not yet available for the diagnosis, prediction or management of Loxosceles spider bites. Therefore, therapeutic options are applied which are not supported by evidence. Still, timely and appropriate support management is crucial in severe cases. An unusual case of viscerocutaneous loxoscelism is presented, also known as systemic loxoscelism, which occurred in a rural area. The case was characterized by severe renal involvement requiring hemodialysis, but its outcome was favorable, despite not using antiloxoscelic serum, which shows the importance of timely management based on appropriate support measures(AU)

Curative effect of intra-tendon sheath hyaluronidase injection guided by ultrasound in treating biceps brachii longitendosynovitis / 中国医师杂志

Objective:To observe the effect of hyaluronidase injection into tendon sheath under muscle bone ultrasound guidance in the treatment of tenosynovitis of long head of biceps brachii.Methods:134 patients with tenosynovitis of the long head of biceps brachii treated in SSL Central Hospital of Dongguan from April 2019 to April 2020 were selected as the research objects. All patients were randomly divided into ordinary injection group and ultrasound-assisted injection group, 67 cases in each group. The general injection group was treated with intra-articular injection of hyaluronidase, and the ultrasound-assisted injection group was treated with intra-tendon sheath injection of hyaluronidase under the guidance of muscle and bone ultrasound. Visual Analog Scales (VAS) score, active flexion joint motion (AFROM), shoulder range of motion (ROM), functional score, flexion strength score, nuclear magnetic resonance imaging (MRI) and local tenosynovitis of the long head of biceps brachii (TLHBB)were measured to evaluate the clinical effect and postoperative complications after treatment.Results:There was no significant difference in gender, age, course of disease, periarthritis of shoulder and disuse atrophy of muscles around shoulder between ordinary injection group and ultrasound-assisted injection group ( P>0.05). After treatment, the AFROM, ROM, function score, forward flexion strength score, and middle wedge angle (MWA) of the two groups were significantly higher than those before treatment ( P<0.05), while the VAS score, humeral head diameter (HHD), biceps long head tendon diameter (BTD), and TLBBB were significantly lower than those before treatment ( P<0.05). The AFROM, ROM, function score, and forward flexion strength score, MWA of the ultrasound-assisted injection group were significantly higher than those of the ordinary injection group ( P<0.05), and the VAS score, HHD, BTD, and TLHBB were significantly lower than those of the ordinary injection group ( P<0.05). The total effective rate of the ultrasound-assisted injection group was higher than that of the ordinary injection group (97.01% vs 85.07%, P<0.05). Conclusions:Intra-tendon sheath hyaluronidase injection guided by ultrasound can effectively treat tenosynouitis of the long head of biceps brachii, relieve shoulder pain and improve shoulder motion.

Uso terapéutico de la hialuronidasa y la triamcinolona en el pseudotumor orbitario / Therapeutic use of hyaluronidase and triamcinolone in orbital pseudotumor

RESUMEN El origen del pseudotumor orbitario no es del todo conocido. Se admite su naturaleza inflamatoria granulomatosa e inespecífica en diferentes localizaciones. El pseudotumor orbitario se define como una respuesta inflamatoria celular pleomórfica, que está usualmente confinado a estructuras de la órbita y tiene una evolución limitada. En este trabajo se presenta una paciente femenina de 16 años, con diagnóstico de pseudotumor orbitario corroborado por biopsia y tomografía axial computarizada, refractaria al tratamiento con esteroides sistémicos, por lo que se decide iniciar con la aplicación de hialuronidasa y triamcinolona en el espacio peribulbar. Los casos agudos casi siempre responden rápidamente al tratamiento con cortocoesteroides, como prednisona, pero debemos tener en cuenta que existen pacientes que son refractarios al tratamiento, por lo que es necesario buscar procedimientos alternativos. Una opción es el uso de hialuronidasa para destruir las uniones extracelulares, y difundir un esteroide de manera local, como la triamcinolona, más efectiva dentro del tejido inflamatorio para provocar un efecto localizado de este. A los tres meses del tratamiento hubo una regresión total del cuadro en esta paciente(AU)

ABSTRACT The exact etiology of orbital pseudotumor is unknown, but its granulomatous unspecific inflammatory nature at various locations has been recognized. Orbital pseudotumor is defined as a cellular pleomorphic inflammatory response of limited evolution often confined to orbital structures. A case is presented of a female 16-year-old patient diagnosed with orbital pseudotumor confirmed by biopsy and computerized axial tomography, refractory to treatment with systemic steroids, due to which it is decided to start treatment with hyaluronidase and triamcinolone in the peribulbar space. Acute cases often respond fast to treatment with corticosteroids such as prednisone. It should be borne in mind that there are patients who are refractory to treatment for whom alternative treatments should be sought. An option is the use of hyaluronidase to destroy extracellular junctions and locally spread a steroid such as triamcinolone, most effectively within the inflammatory tissue to ensure its localized effect. Total regression of the patient's status was observed at three months of treatment(AU)

Randomized, Double-Blind, Placebo-Controlled Trial on the Efficacy of Hyaluronidase in Preventing Perineal Trauma in Nulliparous Women

Dalbergia ecastaphyllum leaf extracts: in vitro inhibitory potential against enzymes related to metabolic syndrome, inflammation and neurodegenerative diseases

For the first time, the anti-hemolytic activity and the enzyme inhibitory activities of Dalbergia ecastaphyllum leaves extracts were tested against α-amylase, α-glucosidase, lipase, acetylcholinesterase, butyrylcholinesterase, tyrosinase and hyaluronidase. The phenolic profile of the obtained extracts was also investigated by high-performance liquid chromatography with photodiode array detection (HPLC-PAD). The extracts showed inhibitory activity against all enzymes evaluated, with the highest inhibitory activity reported for the enzyme hyaluronidase (28.28 ± 2.43 to 72.19 ± 1.40 μg mL-1). The obtained extracts also demonstrate anti-hemolytic activity (52.22 ± 1.62 to 71.17 ± 1.82%). Among the phenolic compounds identified, protocatechuic, vanillic and β-resorcylic acids were the most abundant (1.13 ± 0.06 to 2.53 ± 0.06, 0.90 ± 0.06 to 2.19 ± 0.06 and 1.03 ± 1.62 to 22.11 ± 1.62 mg L-1, respectively). In the statistical analysis, a significant correlation was found between the flavonoids content and all enzymes inhibitory activities. The present study showed that D. ecastaphyllum leaves extracts may have the potential to be used in the therapeutic treatment of several diseases such as Alzheimer, Parkinson, type 2 diabetes mellitus, hyperglycemia, and pigmentation, as well as those associated with oxidative stress.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2)

Background:Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors.Methods:The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed.Results:V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/β)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4.Conclusion:The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.

Effect of hyaluronidase on expression of neuron-glial antigen 2 and myelin basic protein after germinal matrix-intraventricular hemorrhage in premature rabbits / 中华实用儿科临床杂志

Objective@#To investigate the neuroprotective effect of hyaluronidase(HAase) on rabbit brain tissue in germinal matrix-intraventricular hemorrhage(GM-IVH) premature rabbits.@*Methods@#Eighty premature rabbits of gestation age 29 days were randomly divided into normal group, GM-IVH control group and HAase treatment group.The rabbits in GM-IVH control group and HAase treatment group were given intraperitoneal injection of 50 g/L glycerol to establish GM-IVH animal model, while the premature rabbits in normal group were given the same dose of 9 g/L saline.The GM-IVH was screened by using cranial ultrasound.The premature rabbits in HAase treatment group were given HAase into the lateral forebrain while the rabbits in the GM-IVH control group were given the same dose of 9 g/L saline.In 3 days, 7 days and 14 days after birth, the premature rabbits were killed and brain tissue were separated.The expression of neuron-glial antigen 2(NG2) were detected by adopting immunohistochemical method.The expression of myelin basic protein(MBP) were detected by Western blot method.@*Results@#The expression of NG2 in 3 groups of premature rabbits decreased gradually with the increase of age[the values of NG2 in the normal group were(62.65±33.58)×104, (15.61±4.22)×104, and (13.54±4.51)×104 on the 3rd, 7th, and 14th day; those in the GM-IVH control group were (54.58±25.48)×104, (48.91±22.49)×104, (7.18±2.28)×104 on the 3rd, 7th, and 14th day; those in the HAase treatment group were (148.13±27.30)×104, (88.38±14.55)×104, (77.98±18.96)×104 on the 3rd, 7th, and 14th day]. At the same time, the expression of NG2 in the HAase treatment group was higher than that of the GM-IVH control group, and the differences were statistically significant among the 3 groups at any time(all P<0.05). The expression of MBP protein in the 3 groups increased with the increase of day age[MBP protein in the normal group were (0.30±0.22)×103, (1.91±0.43)×103, and (5.67±2.14) ×103 on the 3rd, 7th, and 14th day; those in the GM-IVH control group were (0.87±0.12)×103, (1.15±0.22)×103 and (2.54±0.69) ×103 on the 3rd, 7th, and 14th day; those in the HAase treatment group were (0.91±1.01)×103, (2.25±0.66)×103, (3.40±1.10)×103 on the 3rd, 7th, and 14th day]. The difference was statistically significant between the normal group and the HAase treatment group(P<0.05).@*Conclusions@#After HAase treatment, the expression of NG2 and MBP can be improved to a certain extent, promoting the development of oligodendrocytes and regeneration of myelin sheath.

Construction of engineered Streptococcus zooepidemicus for the production of hyaluronic acid ligosaccharide / 生物工程学报

Hyaluronic acid (HA) is widely used in many fields, such as medicine, cosmetics and food. The bioactivity of HA depends on its molecular weight (Mw). Owing to the important physiological activities and special physiological functions, HA oligosaccharides have important application prospects in medicine fields. Streptococcus zooepidemicus has wide applications in commercial production of HA, due to its short fermentation cycle and strong production intensity. In order to efficiently synthesize HA oligosaccharides and solve the dissolved oxygen in the fermentation process, in this study, we overexpressed HA synthase (HasA) and introduced and optimized the leech hyaluronidase LHAase in Streptococcus zooepidemicus WSH-24. As a result, HA oligosaccharides were efficiently produced with improved dissolved oxygen. After 24 h, HA oligosaccharides production intensity reached to 294.2 mg/(L·h), and the concentration accumulated to 0.97 g/L in flask cultures, which was 1.82 times of the wild strain. Impressively, HA oligosaccharides were increased to 7.06 g/L in 3 L fermentor. The constructed Streptococcus zooepidemicus strain for producing HA oligosaccharides would have broad application prospects.

Effect of hyaluronidase on expression of neuron＿glial antigen 2 and myelin basic protein after germinal ma-trix＿intraventricular hemorrhage in premature rabbits / 中华实用儿科临床杂志

Objective To investigate the neuroprotective effect of hyaluronidase( HAase)on rabbit brain tissue in germinal matrix-intraventricular hemorrhage( GM -IVH)premature rabbits. Methods Eighty premature rabbits of gestation age 29 days were randomly divided into normal group,GM-IVH control group and HAase treatment group. The rabbits in GM -IVH control group and HAase treatment group were given intraperitoneal injection of 50 g/L glycerol to establish GM-IVH animal model,while the premature rabbits in normal group were given the same dose of 9 g/L saline. The GM-IVH was screened by using cranial ultrasound. The premature rabbits in HAase treat-ment group were given HAase into the lateral forebrain while the rabbits in the GM-IVH control group were given the same dose of 9 g/L saline. In 3 days,7 days and 14 days after birth,the premature rabbits were killed and brain tissue were separated. The expression of neuron - glial antigen 2( NG2 )were detected by adopting immunohistochemical method. The expression of myelin basic protei(n MBP)were detected by Western blot method. Results The expression of NG2 in 3 groups of premature rabbits decreased gradually with the increase of ag[e the values of NG2 in the normal group were(62. 65 ± 33. 58)×104(,15. 61 ± 4. 22)×104 ,and(13. 54 ± 4. 51)×104 on the 3rd ,7th ,and 14th day;those in the GM-IVH control group were(54. 58 ± 25. 48)×104 ,(48. 91 ± 22. 49)×104 ,(7. 18 ± 2. 28)×104 on the 3rd ,7th , and 14th day;those in the HAase treatment group were(148. 13 ± 27. 30)×104 ,(88. 38 ± 14. 55)×104 ,(77. 98 ± 18. 96)×104 on the 3rd,7th,and 14th day]. At the same time,the expression of NG2 in the HAase treatment group was higher than that of the GM-IVH control group,and the differences were statistically significant among the 3 groups at any time(all P<0. 05). The expression of MBP protein in the 3 groups increased with the increase of day age[MBP protein in the normal group were(0. 30 ± 0. 22)×103 ,(1. 91 ± 0. 43)×103 ,and(5. 67 ± 2. 14)×103 on the 3rd , 7th,and 14th day;those in the GM-IVH control group were(0. 87 ± 0. 12)×103,(1. 15 ± 0. 22)×103 and(2. 54 ± 0. 69)×103 on the 3rd,7th,and 14th day;those in the HAase treatment group were(0. 91 ± 1. 01)×103 ,(2. 25 ± 0. 66)×103 ,(3. 40 ± 1. 10)×103 on the 3rd,7th ,and 14th day]. The difference was statistically significant between the normal group and the HAase treatment grou(p P<0. 05). Conclusions After HAase treatment,the expression of NG2 and MBP can be improved to a certain extent,promoting the development of oligodendrocytes and regeneration of myelin sheath.

Osmf: Long Term Study Of Treated Patients

Background and Objective: Oral Submucous fibrosis (OSMF) is a chronic debilitating disease and a well recognized premalignant condition of oral cavity. The exact pathogenesis is not well established but the cause is believed to be multifactorial. The most commonly used treatment regimen is combination of Hydrocortisone acetate or Triamcinolone Acetonide with hyaluronidase and recently oral Pentoxifylline, but studies of its long term effect are lacking. The present study was therefore conducted to evaluate the long term effectiveness of afore said drugs. Material & methods: In the present study 60 previously treated patients of OSMF were registered. Out of which 34 patients had received Inj. Hydrocortisone acetate (1.5ml) mixed with Hyaluronidase (1500 I.U.) at weekly interval for 22 weeks, 15 patients had received Inj. Triamcinolone Acetonide (10mg/ml) mixed with Hyaluronidase (1500 I.U.) at biweekly interval for 22 weeks and 11 patients were given Tab Pentoxifylline (Trental) 400mg three times daily for a period of 7 months. Long term outcome was evaluated on the basis of symptom score and sign score. Results: There was statistically significant improvement between long term follow up score and immediate Post-treatment symptom score ( p value < 0.05 ) Interpretation and Conclusion: It was concluded that the improvement in symptoms and signs following treatment increased further or maintained in long term follow up

Biomechanical effect of hyaluronidase on rabbit auricular cartilage / 中华医学美学美容杂志

Objective To evaluate the biomechanical effectiveness of hyaluronidase (HAase) on rabbit auricular cartilage in the early stage.Methods HAase in 3 different concentrations (75 U/ml,150 U/ml,300 U/ml) were injected subcutaneously to rabbit auricle in 3 groups,while normal saline were injected as control.For the comparison of biomechanism properties among different groups and different time,cartilages were harvested at 3rd and 7th day after injection,followed by stress-relaxation assay.Results Auricular cartilage displayed different levels in control group compared with other groups in elastic modulus (P＜0.05) and maxmum stress (P＜0.05) in 3th day as well as 7th day.While both in the same concentration group,there were also differences between the 3th and 7th day (P＜0.05).Conclusions HAase injection can cause changes in biomechanical properties of auricular cartilage.And 7 days would not be enough for the tissue recovery biomechanically.

Comparison of Efficacy of Pentoxifyllin with Standard Regimens Dexamethasone and Hyaluronidase in Patients with Oral Submucous Fibrosis

Aims and objectives The purpose of the study was to clinically evaluate the efficacy of oral pentoxifylline 400mg tablets in comparison to intra-lesional injections of dexamethasone (4 mg/ml), hyaluronidase 1, 500 IU and 0.5 ml of lignocaine 2% and a combination of the two in the management of oral sub-mucous fibrosis (OSF) patients. Materials and methods The study population consisted of randomly selected 75 patients with OSF. Patients were divided into three groups. A total of 25 patients were allocated each in dexamethasone group, dexamethasone and pentoxifylline group and pentoxifylline group. Dexamethasone group received weekly intra-lesional injections of dexamethasone (4 mg/ml), hyaluronidase 1, 500 IU and 0.5 ml of lignocaine 2% for a period of 12 weeks. Pentoxifylline group received oral pentoxifylline 400-mg tablets thrice daily for 12 weeks. Dexamethasone and pentoxifylline group received both the drugs. Parameters taken in the study were burning sensation and mouth opening. Results In the present study, improvement in the patients symptomatic parameters were statistically significant (p < 0.001) within groups (intra-group comparison), but not significant (p > 0.001) when compared between the groups (inter-group comparison). Conclusion Study found that the parameters like burning sensation and mouth opening showed statistically significant improvement for all the drug groups. The drug pentoxifylline showed similar improvement in the clinical parameters as that of dexona. For this reason, pentoxifylline can be indicated as a good alternative treatment for OSF in patients in whom dexamethasone is contraindicated, or those who cannot make frequent visits for intra-lesional injections.

Highlights in the knowledge of brown spider toxins

Abstract Brown spiders are venomous arthropods that use their venom for predation and defense. In humans, bites of these animals provoke injuries including dermonecrosis with gravitational spread of lesions, hematological abnormalities and impaired renal function. The signs and symptoms observed following a brown spider bite are called loxoscelism. Brown spider venom is a complex mixture of toxins enriched in low molecular mass proteins (440 kDa). Characterization of the venom confirmed the presence of three highly expressed protein classes: phospholipases D, metalloproteases (astacins) and insecticidal peptides (knottins). Recently, toxins with low levels of expression have also been found in Loxosceles venom, such as serine proteases, protease inhibitors (serpins), hyaluronidases, allergen-like toxins and histamine-releasing factors. The toxin belonging to the phospholipase-D family (also known as the dermonecrotic toxin) is the most studied class of brown spider toxins. This class of toxins single-handedly can induce inflammatory response, dermonecrosis, hemolysis, thrombocytopenia and renal failure. The functional role of the hyaluronidase toxin as a spreading factor in loxoscelism has also been demonstrated. However, the biological characterization of other toxins remains unclear and the mechanism by which Loxosceles toxins exert their noxious effects is yet to be fully elucidated. The aim of this review is to provide an insight into brown spider venom toxins and toxicology, including a description of historical data already available in the literature. In this review article, the identification processes of novel Loxosceles toxins by molecular biology and proteomic approaches, their biological characterization and structural description based on x-ray crystallography and putative biotechnological uses are described along with the future perspectives in this field.

Primary cultures of mouse small intestinal epithelial cells using the dissociating enzyme type I collagenase and hyaluronidase

The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.

Highlights in the knowledge of brown spider toxins

Abstract Brown spiders are venomous arthropods that use their venom for predation and defense. In humans, bites of these animals provoke injuries including dermonecrosis with gravitational spread of lesions, hematological abnormalities and impaired renal function. The signs and symptoms observed following a brown spider bite are called loxoscelism. Brown spider venom is a complex mixture of toxins enriched in low molecular mass proteins (4-40 kDa). Characterization of the venom confirmed the presence of three highly expressed protein classes: phospholipases D, metalloproteases (astacins) and insecticidal peptides (knottins). Recently, toxins with low levels of expression have also been found in Loxosceles venom, such as serine proteases, protease inhibitors (serpins), hyaluronidases, allergen-like toxins and histamine-releasing factors. The toxin belonging to the phospholipase-D family (also known as the dermonecrotic toxin) is the most studied class of brown spider toxins. This class of toxins single-handedly can induce inflammatory response, dermonecrosis, hemolysis, thrombocytopenia and renal failure. The functional role of the hyaluronidase toxin as a spreading factor in loxoscelism has also been demonstrated. However, the biological characterization of other toxins remains unclear and the mechanism by which Loxosceles toxins exert their noxious effects is yet to be fully elucidated. The aim of this review is to provide an insight into brown spider venom toxins and toxicology, including a description of historical data already available in the literature. In this review article, the identification processes of novel Loxosceles toxins by molecular biology and proteomic approaches, their biological characterization and structural description based on x-ray crystallography and putative biotechnological uses are described along with the future perspectives in this field.(AU)