Expression, purification, and functional verification of recombinant human glycoprotein hormone beta 5/alpha 2 fusion protein in CHO-S cells / 中国药理学通报

Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice.

Effect of suspension combined with acupuncture on chronic non-specific neck pain / 中国康复理论与实践

ObjectiveTo explore the effect of suspension combined with acupuncture on chronic non-specific neck pain (CNNP). MethodsA total of 81 patients with unilateral CNNP from the rehabilitation outpatient clinic of Wangjing Hospital from March, 2022 to March, 2023 were divided into suspension group (n = 27), acupuncture group ((n = 27) and combined group (n = 27) randomely. The suspension group underwent conventional suspension, the acupuncture group received conventional acupuncture treatment, and the combined group underwent suspension and acupuncture, for six weeks. The Visual Analogue Scale (VAS), Neck Disability Index (NDI), and cross-sectional area (CSA) of longus colli and multifidus in the neck using ultrasound imaging were compared before and after treatment. ResultsAfter treatment, the VAS and NDI score, and CSA of longus colli and multifidus improved in three groups (|t| > 4.473, P < 0.001), and the combined group was better than the other two groups (P < 0.05). ConclusionSuspension and acupuncture can relief the pain, improve function, and increase CSA of longus colli and multifidus in patients with unilateral CNNP, while the combination of the two methods is better.

Clinical application of 8-0 polypropylene scleral-sutured fixed intraocular lens suspension implantation with the double knots technique in aphakic eyes / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the clinical efficacy of 8-0 polypropylene scleral-sutured fixed intraocular lens(IOL)suspension implantation with the double knots technique in aphakic eyes.METHODS: Retrospective case series study. The data of 30 aphakic cases(31 eyes, 22 males)that underwent IOL suspension in our hospital from January 2021 to November 2022 were collected. The suspension of IOL(AcrySof IQ or Tecnis ZCB00)was performed by 8-0 polypropylene scleral-sutured with the double knots technique. The visual acuity, intraocular pressure(IOP), IOL position and complications with at least 6 mo of follow-up were observed.RESULTS: The mean preoperative uncorrected visual acuity(UCVA, LogMAR)and best-corrected visual acuity(BCVA, LogMAR)were 2.53±0.78 and 0.35±0.26, respectively, which were 0.58±0.26 and 0.36±0.27 at 6 mo postoperatively, respectively. And the differences in UCVA were statistically significant(t=15.408, P&#x003C;0.01), whereas the difference in BCVA was not(t=-1.677, P=0.104). There were no intraoperative complications, with IOL position all centered, but 3 eyes had IOL tilt, 2 eyes had intraocular hypertension, 5 eyes had corneal edema, and 1 eye had suture exposure postoperatively. There were no complications such as hyphema, vitreous hemorrhage, macular edema, corneal endothelial decompensation, hypotony, choroidal detachment, retinal detachment, fulminant superior choroidal hemorrhage, endophthalmitis, or others.CONCLUSION: The 8-0 polypropylene scleral-sutured fixed intraocular lens suspension implantation with the double knots technique can improve the postoperative visual acuity of aphakic patients, and fewer complications, which is an option for the treatment of aphakia, dislocation of the lens and ligament abnormalities.

Surgical and Functional Outcome of Laparoscopic Vaginal Suspension Combined with Suture Rectopexy for Management of Pelvic Organ Prolapse

Abstract Purpose Laparoscopic techniques to treat pelvic organ prolapse are gaining popularity around the globe due to their low recurrence rates and better functional results compared to perineal techniques. However, the optimum surgical procedures are not yet determined. In the current research, we suggest a novel surgical approach, laparoscopic vaginal suspension with suture rectopexy, to treat multiorgan pelvic prolapse. Methods This prospective cohort trial was conducted from March 2018 to March 2022 and comprised 35 females with multiorgan pelvic organ prolapse with obstructed defecation symptoms. A residual rectal prolapse was still present despite the manual reduction of uterine prolapse. Patients' conditions before and after the operation were monitored regarding the obstructed defecation score, sexual function, need for laxatives, anorectal manometry pressures, anorectal sensations, and recurrence. The mean follow-up duration was one year. Results Modified Longo score for obstructed defecation significantly decreased at six and twelve months after surgery. Additionally, a significant reduction was reported in the number of patients who needed laxatives at six and twelve months after surgery. Anorectal manometry pre- and post-surgery showed a significant elevation in the mean squeeze pressure and a decline in all rectal sensations. All parameters of the female sexual function scoring system increased postoperatively. No recurrence was reported during follow-up. Conclusion For multiorgan pelvic prolapse, laparoscopic vaginal suspension combined with suture rectopexy has excellent functional outcomes, minimal morbidity, and low cost.

Clinical study of modified suspension reduction method combined with percutaneous vertebroplasty in the treatment of thoracolumbar osteoporotic compression fracture / 中国骨伤

OBJECTIVE@#To investigate the clinical effect of modified suspension reduction method combined with percutaneous vertebroplasty in the treatment of osteoporotic thoracolumbar compression fractures.@*METHODS@#From February 2020 to October 2021, 92 patients with thoracolumbar osteoporotic compression fracture were treated by percutaneous vertebroplasty. According to different treatment methods, they were divided into the observation group and the control group. The observation group was treated with modified suspension reduction and then percutaneous vertebroplasty, while the control group was treated with percutaneous vertebroplasty alone. The observation group (47 cases), including 20 males and 27 females, the age ranged from 59 to 76 years old with an average of (69.74±4.50) years old, fractured vertebral bodies:T10(2 cases), T11(7 cases), T12(19 cases), L1(14 cases), L2(5 cases);the control group(45 cases), including 21 males and 24 females, the age ranged from 61 to 78 years old with an average of (71.02±3.58) years old, fractured vertebral bodies:T10(3 cases), T11(8 cases), T12(17 cases), L1(12 cases), L2(5 cases);The leakage of bone cement were observed, the visual analogue scale (VAS), Oswestry lumbar dysfunction index (ODI), anterior vertebrae height (AVH), Cobb angle of kyphosis and the amount of bone cement injected before and after operation were recorded and compared between the two groups.@*RESULTS@#All patients were followed up, ranged from 6 to10 with an average of (8.45±1.73) months. Two patients ocurred bone cement leakage in observation group and 3 patients in control group. AVH of observation group increased (P<0.05) and Cobb angle of injured vertebrae decreased (P<0.05). Cobb angle of injured vertebrae and AVH of the control group were not significantly changed (P>0.05). Cobb angle of injured vertebrae of the observation group was lower than that of control group (P<0.05) and AVH was higher than that of the control group (P<0.05). In the observation group, VAS before operation and 1 week, 3 and 6 months after operation respectively were(7.32±1.05) scores, (3.56±1.18) scores, (1.83±0.67) scores, (1.27±0.34) scores, and ODI were(40.12±14.69) scores, (23.76±10.19) scores, (20.15±6.39) scores, (13.45±3.46) scores. In the control group, VAS before operation and 1 week, 3 and 6 months after operation respectively were(7.11±5.26) scores, (3.82±0.68) scores, (1.94±0.88) scores, (1.36±0.52) scores, and ODI were(41.38±10.23) scores, (25.13±14.22) scores , (20.61±5.82) scores, (14.55±5.27) scores . The scores of VAS and ODI after operation were lower than those before operation (P<0.05), but there was no significant difference between the two groups (P<0.05).@*CONCLUSION@#Modified suspension reduction method combined with PVP surgery for osteoporotic thoracolumbar compression fractures has achieved good clinical results, which can effectively relieve lumbar back pain, restore vertebral height, correct kyphosis, improve lumbar function and patients' quality of life.

Fractura de mandíbula en pacientes parcialmente edéntulos. Alternativa de tratamiento cerrado. Serie de casos / Mandibular fractures in partially edentulous patients. Closed treatment alternatives. A Cases series

Para el tratamiento de fracturas mandibulares existen protocolos cuyo propósito primordial es la función temprana. Existen diversos motivos por los cuales no se puede ejecutar ciertas técnicas en Venezuela, principalmente por problemas económicos para la adquisición de materiales de osteosíntesis especializados. Por esta razón, se ha recurrido a técnicas quirúrgicas antiguas. Se presenta un estudio con diseño longitudinal de tipo descriptivo, para el reporte de serie de 5 casos clínicos de fracturas de mandíbula, con los siguientes criterios de inclusión: presentar fractura mandibular en pacientes dentados parcialmente con imposibilidad al acceso al material de osteosíntesis del sistema de cargas soportadas. Tratados bajo procedimiento quirúrgico cerrado, la condición de edentulismo parcial confiere a la fractura de mandíbula inestabilidad, dificulta la reducción anatómica, pérdida de dimensión vertical y transversal, las que se recuperan a través de la elaboración de férulas tipo Gunning modificadas. Estas férulas se mantuvieron en posición con alambrados de suspensión ósea tipo circummandibulares y circumzigomáticos, como medios para establecer estabilidad en el tiempo. Se obtuvieron resultados satisfactorios, por lo que, aún hoy en día se pueden plantear como opciones de tratamiento.

For the treatment of mandibular fractures, there are protocols whose primary purpose is early function. However, several reasons prevent the execution of these techniques in Venezuela, mainly due to economic problems regarding the acquisition of specialized osteosynthesis materials. As a result, there is a necessity to resort to old surgical techniques. A study with a descriptive longitudinal design is presented, reporting 5 clinical cases of jaw fractures with the following inclusion criteria: presenting a mandibular fracture in partially dentate patients with impossibility of accessing the osteosynthesis material of the load-bearing system. The treatment conducted under closed surgical procedures. The condition of partial edentulism confers instability to the mandible fracture, hindering anatomical reduction and causing a loss of vertical and transverse dimension. These issues are addressed through the utilization of modified Gunning-type splints, which help in recovery. The splints were maintained in position with circum-mandibular and circum-zygomatic bone suspension wiring as a means to establish stability over time. Satisfactory results were obtained, indicating that these techniques can still be considered as treatment options today.

Comparative study of the efficacy and safety of two grafting procedures (an automated epidermal harvesting system and non-cultured epidermal cell suspension) in the treatment of stable vitiligo

Background: The non-cultured epidermal cell suspension method is a well-established but tedious grafting modality in the management of stable vitiligo. Recently a more user-friendly automated epidermal harvesting system has been introduced. Aim: This was a pilot study to compare the efficacy and safety outcomes of the above two grafting procedures. Study design: The study was a single-blinded split-body randomised controlled trial. After scientific and ethical clearance, the trial was registered with CTRI (CTRI/2018/05/014225). Thirty consenting patients of stable vitiligo with 60 near-symmetrical patches were recruited. Block randomisation was done using computer-generated randomisation software and each patch was allocated either of the two grafting modalities. Efficacy was assessed by the Physician Global Assessment Scale on serial images and pain by the Numerical Rating Pain Scale. Results and conclusion: The non-cultured epidermal cell suspension was found to be an overall statistically superior technique to the automated epidermal harvesting system in terms of efficacy (re-pigmentation). Both donor and recipient site complications were significantly less with the automated epidermal harvesting system grafting and this method had the distinct advantage of being a painless and easy technique with minimal recovery time. A novel observation was that a good colour match and near-complete re-pigmentation occurred in patients with a darker skin colour with both techniques. Limitations: The main limitation of our study was the small sample size. Also, the size of the treated patches was limited such that they could be covered by the 5 × 5 cm size of the automated epidermal harvesting system blade. However, a larger area can be covered with multiple sessions.

Pachyoliva columellaris G.B. Sowerby I, 1825 (Olividae, Caenogastropoda) in Peru: tidal migrations and population density

The gastropod Pachyoliva columellaris (formerly Olivella columellaris) is particularly common on many sandy beaches in North Peru. In the studied region north of Paita, Piura province, the species reaches densities of over 80,000 individuals per beach meter. The snails perform tidal migrations throughout the lunar cycle although tidal amplitudes differ four-fold between neap and spring tide. Not all animals complete their tidal migrations, and significant numbers remain above the water line at low tide. Throughout the tidal cycle, the proportion of small individuals is increased near the water line whereas large animals dominate in lower beach zones. I conclude that the sandy beaches north of Paita are well-suited for further studies of P. columellaris and the mechanisms underlying tidal migrations in invertebrates.

El gasterópodo Pachyoliva columellaris (antes Olivella columellaris) es particularmente común en las playas arenosas del norte de Perú. En la región estudiada al norte de Paita, provincia de Piura, la especie alcanza densidades de más de 80.000 individuos por metro de playa. Los caracoles realizan migraciones mareales durante todo el ciclo lunar, aunque las amplitudes de marea difieren cuatro veces entre la marea muerta y la marea viva. No todos los animales completan sus migraciones mareales y un número significativo permanece por encima del nivel del agua en marea baja. Durante todo el ciclo de marea, la proporción de individuos pequeños aumenta cerca del nivel del agua, mientras que los animales grandes dominan en las zonas bajas de la playa. Concluyo que las playas arenosas al norte de Paita son apropiadas para estudios adicionales de P. columellaris y los mecanismos de las migraciones de marea en invertebrados.

Suspensión frontal con aponeurosis temporal en paciente con ptosis palpebral traumática recidivante crónica / Frontal suspension with temporal aponeurosis in a patient with chronic recurrent traumatic eyelid ptosis

La ptosis palpebral es una de las patologías más frecuentes en la consulta de oftalmología, tanto en urgencias como en el ámbito ambulatorio. El trauma del párpado superior puede provocar ptosis o retracción o una combinación de ambos. En este tipo de ptosis palpebral, su resolución mediante tratamiento quirúrgico, existiendo múltiples alternativas de procedimientos que se decidirá de acuerdo con la causas que las originan, así como la severidad de la ptosis. En este trabajo es a propósito de un caso clínico en el cual nos encontramos una paciente con ptosis palpebral traumática recidivante, utilizándose suspensión frontal con aponeurosis de músculo temporal, realizándose una variación de la técnica de Crawford disminuyendo la probabilidad de nueva recidiva

Palpebral ptosis is one of the most frequent pathologies in the ophthalmology consultation, both in the emergency room and in the outpatient setting Trauma to the upper eyelid can cause ptosis or retraction or a combination of both. In this type of eyelid ptosis its resolution by surgical treatment, there are multiple alternatives of procedures that will be decided according to the causes that originate them, as well as the severity of the ptosis, in this work is about a clinical case in which we find a patient with Recurrent Traumatic Palpebral Ptosis, using frontal suspension with aponeurosis of temporal muscle, performing a variation of the Crawford Technique decreasing the probability of recurrence.

Stability of sulfadiazine sugar-free oral suspensions for the treatment of congenital toxoplasmosis

Abstract This study aimed to develop and evaluate the stability of sulfadiazine sugar-free extemporaneous oral suspensions, focusing on treating congenital toxoplasmosis. The excipients were carefully chosen to obtain safe products for the pediatric population. Sulfadiazine suspensions (100 mg/ mL) were prepared from the raw material or tablets, stored in amber glass bottles at 5±3ºC, and evaluated at 0, 14, and 30 days. An ultra-performance liquid chromatographic method was developed and validated to assay the drug. The particle size ranged from 29.3 to 50.6 µm, with some variation over the study; pH values around 7.0 and non-Newtonian behavior were observed without modification in the period. Formulations showed a fast dissolution rate (>80% in 15 minutes) without variation over the study. The drug assay was about 100% of the label claimed throughout the study, demonstrating the chemical stability and the preparations' dose homogeneity. The microbiological investigation indicated that both preparations met the requirements for the microbial count and absence of pathogens. In conclusion, the developed formulations can be used for 30 days when stored under refrigeration. The oral suspensions produced are easy to prepare and contain safe components, providing an alternative for congenital toxoplasmosis treatment in children.

Suspensión de antirresortivos en osteonecrosis asociada a medicamentos: revisión de la literatura / Suspension of antiresorptive drugs in medication-related osteonecrosis of the jaws: a review of the literature

La suspensión de antirresortivos en el contexto de cuadros de osteonecrosis asociada a medicamentos (ONAM) es controversial. Aunque la evidencia de ca-lidad al respecto es insuficiente, múltiples socieda-des profesionales y grupos de trabajo han sugerido emplear este recurso. Recientemente la Sociedad Americana de Cirujanos Orales y Maxilofaciales ha puesto en duda sus beneficios. En el presente estudio abordamos esta temática en dos situaciones clínicas diferentes. Por un lado, analizamos la suspensión de los antirresortivos en pacientes asintomáticos an-tes de llevar a cabo procedimientos invasivos en los maxilares para disminuir el riesgo de desarrollo de ONAM. Por otro lado, evaluamos la suspensión de los antirresortivos en pacientes con ONAM establecida para mejorar el pronóstico de la enfermedad (AU)

The suspension of antiresorptive drugs in the context of medication-related osteonecrosis of the jaws (MRONJ) is controversial. Despite the lack of quality evidence, several professional associations and working groups have made suggestions in using this resource. Recently the American Association of Oral and Maxillofacial Surgeons has questioned its benefits. In the present study we address this issue in two different clinical situations. On the one hand, we analyzed the suspension of antiresorptive agents in asymptomatic patients before carrying out invasive procedures in the jaws to reduce the risk of developing MRONJ. On the other hand, we evaluated the suspension of antiresorptive drugs in patients actually presenting MRONJ to improve the prognosis of the disease (AU)

Comparison and improvement of palatability for multisourced oral azithromycin suspension / 药学学报

Azithromycin dry suspension is one of the most commonly used drugs in pediatric clinic, but its taste masking has been difficult to achieve. 5 representative products of azithromycin dry suspension were chose to compare their tastes both using electronic tongue and human sensory evaluation methods, and there existed the differences of bitterness, later bitterness, graininess, and adhesion among these products. Raman micro-imaging was used to determine the difference in taste mainly due to different prescription ingredients and manufacturing techniques. Through mixing the dry suspensions with alkaline mixing solvent, the bad taste of each product was masked after evenly dispersing in the solvent, and their tastes were all close to the taste of the solvent. In the future, it is planned to investigate the stability and bioavailability of the solvent preparations, and then to give the medication suggestion of solvent preparation after ensuring their efficacy.

Development of a nature posture treatment suspension therapy based on ICF-CY / 中国康复理论与实践

ObjectiveTo develop a rehabilitation program of nature posture treatment (NPT) suspension therapy based on the International Classification of Functioning, Disability and Health-Children and Youth version (ICF-CY) framework, and apply it to neurodevelopmental disorders. MethodsThe ICF-CY theoretical group (group A) and NPT suspension therapy group (group B) were established. Group A searched literature from common databases, to extract high-frequency words related to suspension therapy and match with categories of ICF-CY, to develop ICF-CY theoretical framework of the NPT suspension therapy. Group B developed specific rehabilitation procedures and training items based on the framework to compose the training pool. A total of 110 children aged less than six years with neurodevelopmental disorders and associated motor impairments were selected from outpatient or inpatient of the First Affiliated Hospital of Xinxiang Medical University, between October, 2019 and October, 2022. They were randomly divided into control group (n = 55) and clinical group (n = 55), who received routine neurodevelopmental therapy and NPT suspension therapy program based on ICF-CY, respectively, for a week. The incidence of satisfaction, acceptance and adverse events were observed. ResultsTwo cases in the control group and four cases in the clinical group dropped down. For the clinical group, the incidence of satisfaction was 98% (50/51), with acceptance of 96% (49/51), and one adverse event occurred. For the control group, the incidence of both the satisfaction and acceptance was 100%, and no adverse event occurred. There was no significant difference in the incidence of satisfaction, acceptance and the adverse event (P > 0.05). ConclusionThe NPT suspension therapy program based on the ICF-CY framework is safe and acceptable for children with neurodevelopmental disorders.

LC-MS analysis of 2-(2-phenylethyl) chromones in sodium chloride-treated suspension cells of Aquilaria sinensis / 中国中药杂志

Qualitative and quantitative analysis of 2-(2-phenylethyl) chromones in sodium chloride(NaCl)-treated suspension cells of Aquilaria sinensis was conducted by UPLC-Q-Exactive-MS and UPLC-QQQ-MS/MS. Both analyses were performed on a Waters T3 column(2.1 mm×50 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as mobile phases at gradient elution. MS data were collected by electrospray ionization in positive ion mode. Forty-seven phenylethylchromones was identified from NaCl-treated suspension cell samples of A. sinensis using UPLC-Q-Exactive-MS, including 22 flindersia-type 2-(2-phenylethyl) chromones and their glycosides, 10 5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones and 15 mono-epoxy or diepoxy-5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones. Additionally, 25 phenylethylchromones were quantitated by UPLC-QQQ-MS/MS. Overall, the rapid and efficient qualitative and quantitative analysis of phenylethylchromones in NaCl-treated suspension cells of A. sinensis by two LC-MS techniques, provides an important reference for the yield of phenylethylchromones in Aquilariae Lignum Resinatum using in vitro culture and other biotechnologies.

Clinical efficacy of small incision conjoint fascial sheath suspension in treatment of severe congenital blepharoptosis / 中华医学美学美容杂志

Objective:To analyze the clinical effect of small incision approach conjoint fascial sheath (CFS) suspension in the treatment of congenital severe blepharoptosis, and to discuss its advantages and disadvantages compared with conventional CSF suspension.Methods:From February 2020 to August 2022, 42 cases of severe blepharoptosis in the Department of Burn, Plastic and Cosmetic Surgery, Shaanxi Provincial People's Hospital were divided into the observation group (23 cases, 39 eyes) and the control group (19 cases, 37 eyes). The observation group was treated with small incision CFS suspension surgery, while the control group was treated with conventional CFS suspension surgery. The correction effect, complications, recovery time and other conditions between the two groups at different times after surgery were compared.Results:During postoperative follow-up at 1 week, 1 month, 3 months, and 6 months, there was no significant difference in the corrective effect between the two surgical methods at each time point (all P>0.05). The incidence of complications in the observation group at each time point that was 26.3%, 15.7%, 10.5%, and 5.2%, respectively, while the incidence of complications in the control group was 60.0%, 20.0%, 14.2%, and 8.6%, with statistical differences in the first week after surgery (χ 2=8.74, P=0.011). The average postoperative swelling time in the observation group was 4.2 days, which was less than 5.8 days in the control group. During a 6-month follow-up, it was found that there was a decrease in scar hyperplasia in the observation group of 9.1% (2/22) compared to the control group of 16.7% (3/18) (χ 2=0.023, P=0.878). The difference was of no statistical significance. Conclusions:CFS suspension with small incision in the treatment of moderate and severe blepharoptosis has the advantages of ideal correction effect, small damage range, and few postoperative complications, but the operation area is small, the operation is difficult, and the surgeon has higher requirements.

Expression, purification and functional verification of recombinant human α-galactosidase A in suspension CHO-S / 中国药理学通报

Aim To express and purify rhα-Gal A with a 6 X His tag via using a serum-free expression system in high-density suspension culture of Chinese hamster ovary cells ( CHO-S) , and to verify the scavenging effect of rhα-Gal A on globular trisaccharide ceramide (Gb3 or GL3) . Methods The construction of recombinant protein expression vector, pcDNA4-GLA, was achieved by fusing the human α-galactosidase cDNA, gla, with 6 X His tag and artificial DNA synthesis. The expression plasmid was transfected into the suspended CHO-S to express rhα-Gal A and then purified. Following this procedure, we determined rhα-Gal A's expression, the enzymatic activity, and the glycosylation of the recombinant enzyme. Co-incubation with cultured cells was performed to examine whether rhα-Gal A could be taken up into the cells and effectively remove Gb3 substrates. Results rhα-Gal A was successfully expressed and purified after transiently transfecting pcDNA4-GLA into the suspended CHO-S, and the yield was up to (100 ±20. 6) mg • L

Cerebrospinal fluid inflammatory cytokine profiles of patients with neurotropic parasitic infections

@#The pathogenesis of chronic parasitic central nervous system (CNS) infections, including granulomatous amoebic meningoencephalitis (GAE), cerebral toxoplasmosis (CT), and neurocysticercosis (NCC), is primarily due to an inflammatory host reaction to the parasite. Inflammatory cytokines produced by invading T cells, monocytes, and CNS resident cells lead to neuroinflammation which underlie the immunopathology of these infections. Immune molecules, especially cytokines, can therefore emerge as potential biomarker(s) of CNS parasitic infections. In this study, cerebral spinal fluid (CSF) samples from suspected patients with parasitic infections were screened for pathogenic free-living amoebae by culture (n=2506) and PCR (n=275). Six proinflammatory cytokines in smear and culture-negative CSF samples from patients with GAE (n = 2), NCC (n = 7), and CT (n = 23) as well as control (n = 7) patients were measured using the Multiplex Suspension assay. None of the CSF samples tested was positive for neurotropic free-living amoebae by culture and only two samples showed Acanthamoeba 18S rRNA by PCR. Of the six cytokines measured, only IL-6 and IL-8 were significantly increased in all three infection groups compared to the control group. In addition, TNFa levels were higher in the GAE and NCC groups and IL-17 in the GAE group compared to controls. The levels of IL-1b and IFNg were very low in all the infection groups and the control group. There was a correlation between CSF cellularity and increased levels of IL-6, IL-8, and TNFa in 11 patients. Thus, quantifying inflammatory cytokine levels in CSF might help with understanding the level of neuroinflammation in patients with neurotropic parasitic diseases. Further studies with clinico-microbiological correlation in the form of reduction of cytokine levels with treatment and the correlation with neurological deficits are needed.

Bovine viral diarrhea virus Erns protein expressed in Chinese hamster ovary cells and its immunogenicity analysis / 生物工程学报

The aim of this study was to produce Erns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified Erns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-Erns was constructed based on the gene sequence of BVDV-1 NADL strain. The Erns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-Erns into CHO cells. The expression and purification of the Erns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the Erns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified Erns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the Erns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the Erns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log10 was induced in rabbits. In this study, purified BVDV Erns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.

Effect of different suspension moxibustion methods on syndrome characteristics of rats with rheumatoid arthritis of heat bi syndrome based on "moxibustion can be used for heat syndrome" / 中国针灸

OBJECTIVE@#To observe the effects of different suspension moxibustion methods on the syndrome characteristics and inflammatory factors of rats with rheumatoid arthritis (RA) of heat bi syndrome and to prove the concept of "moxibustion can be used for heat syndrome".@*METHODS@#Among seventy Wistar rats, 12 rats were randomly selected as a normal group, and the remaining rats were induced by collagen combined with wind, dampness, and heat environmental stimulation to establish the RA model of heat bi syndrome. Forty-eight rats with successful model establishment were further randomly divided into a model group and three moxibustion groups (mild moxibustion group, rotating moxibustion group and sparrow-pecking moxibustion group), with 12 rats in each group. The acupoints "Quchi" (LI 11), "Dazhui" (GV 14) and ashi point were used in all moxibustion groups, with mild moxibustion, rotating moxibustion, and sparrow-pecking moxibustion intervention given respectively, each acupoint was treated with moxibustion for 10 min a day, and 6 days were considered one course of treatment, with a total of three courses. After the intervention, the arthritis index (AI), the Evans blue (EB) extravasated volume in the soft tissue of the right hind paw, and the levels of tumor necrosis factor (TNF)-α and interleukin (IL)-10 in the serum were measured by ELISA in each group. The volume of the bilateral hind paw was measured; the infrared thermal imaging was collected to analyze the temperature of the plantar area of the bilateral foot pads, and the reaction time of plantar heat pain was calculated before and after modeling, as well as after the 1st, 2nd and 3rd courses of interrention. The ankle dorsiflexion angle of the right hind foot was also measured before and after modeling, as well as after the intervention.@*RESULTS@#After modeling, compared with the normal group, the rats in the model group had more high-temperature areas in the bilateral hind limbs, abnormal AI score, abnormal bilateral hind paw volume, abnormal temperature of the plantar area of the bilateral foot pads, abnormal foot pain response time, abnormal right hind ankle dorsiflexion angle, abnormal right hind paw soft tissue EB extravasation, and abnormal serum TNF-α and IL-10 levels (P<0.01, P<0.05). After the intervention, compared with the model group, the rats in each moxibustion group had decreased or disappeared high-temperature areas in the bilateral hind limbs, EB extravasated volume in the soft tissue of the right hind paw was reduced (P<0.05), and the right ankle dorsiflexion angle was increased (P<0.05), serum level of TNF-α was reduced, and level of IL-10 increased (P<0.05); the AI scores in the mild moxibustion group and the sparrow-pecking moxibustion group was decreased (P<0.01, P<0.05). After the 1st, 2nd and 3rd courses of intervention, compared with the model group, the bilateral hind paw volume of rats in each moxibustion group was decreased (P<0.05, P<0.01), and plantar heat pain reaction time was increased (P<0.05). After the 2nd course and the 3rd course of intervention, the temperature of the right hind paw pad area was decreased in each moribustion group (P<0.05); after the 3rd courses of intervention, the temperature of the left hind paw pad area was decreased in the mild moxibustion group (P<0.05).@*CONCLUSION@#Suspension moxibustion could adjust the serum levels of TNF-α and IL-10 to improve the syndrome characteristics of RA rats of heat bi syndrome, such as joint redness, swelling, heat, pain and activity restriction. The effect of mild moxibustion is the most prominent. The findings could provide scientific basis for "moxibustion can be used for heat syndrome".

Survey on the Status of Descriptions regarding Reference Information for Dispensing and Medication Support in the Interview Form / 医薬品情報学

Objective: We examined the descriptions of reference information regarding the dispensing process in the interview form (IF). If there was no description, we determined the test data’s availability from the pharmaceutical company.Methods: The survey targeted 78 drugs that complied with the new description IF guidelines, for regular tablets and capsules launched from April 2018 to December 2021. The survey was conducted between March 13th and June 14th, 2022. We investigated the reference information regarding the stability of the medications during the crushing and simple suspension method during dispensing.Results: Regarding the data related to the stability of medications during the crushing process, ten drugs were described as having “data available,” 18 drugs were to be “inquired individually,” 31 drugs had “no applicable data,” and 14 drugs had “no items.” Regarding the status of descriptions on the simple suspension method, ten drugs were described as having “data available,” 18 drugs had data that was to be “inquired individually,” 32 drugs had “no applicable data,” and 14 drugs had “no items.” Regarding the 35 drugs for which both the stability during crushing and the simple suspension method were listed as either “no applicable data” or “no items,” we contacted the pharmaceutical companies to inquire about the test data for the stability during the crushing and simple suspension method. We found that four drugs had “data available” on the stability of medications during the crushing process, while six drugs had “data available” on the simple suspension method.Conclusion: Many IFs that complied with the new description guidelines had items for reference information regarding dispensing. However, the test data’s description was not sufficient. Furthermore, even if there is no description of reference information regarding dispensing in the IF, we confirmed that the pharmaceutical company owns the test data.