In this study, we constructed a GLP-2R-HEK293 cell line and established a method for the determination of the in vitro biological activity of teduglutide based on HTRF, after optimizing experimental conditions and methodological verification. We also carried out relative potency detection of teduglutide pharmaceutical products using this method. The result showed that there was a quantitative-efficient relationship between the teduglutide activity and cAMP contents in GLP-2R-HEK293 cells, which conformed to four-parameter model. Method verification results of five concentrations of teduglutide (64%, 80%, 100%, 125% and 156%) met the requirements of the General Rules of Chinese Pharmacopoeia, 2020 edition, Volume IV (9401). We then analyzed the relative potency of three batches of teduglutide drug substances and three batches of drug products. The linearity, regression and parallelism of the obtained curves all fit the system suitability requirements. The relative potency of six batches of teduglutide was from 83% to 105%. In summary, the biological activity detection method established in this study was accurate, precise, simple and time-saving, which can be used for quality control of teduglutide pharmaceutical products.

Objective To investigate the advantages and efficacy of traction with titanium clips in endoscopic submucosal dissection(ESD)for large laterally spreading tumor(LST)in rectum and sigmoid colon.Methods 67 patients with large sigmoid or rectal LST underwent ESD from January 2018 to June 2022 were analyzed retrospectively,including 32 patients in Group A and 35 patients in Group B.Group A was treated with clip-line traction and group B was treated with traditional ESD.The size of lesion,the total operation time,the submucosal dissection time,submucosal dissection rate,submucosal injection number,en bloc resection rate,R0 resection rate,curative resection rate and complications of the two groups were compared.Results LST-G-M was the most common type and villous adenoma was the main pathology in both groups.There were no differences in en bloc resection rate,R0 resection rate and incidence of complications between the two groups.The average size of group A was(13.6±8.4)cm2,significantly larger than that in group B(9.3±4.7)cm2,the total operation time was(42.3±10.3)min in group A,significantly shorter than that in group B(47.9±10.1)min,submucosal dissection time was(30.7±8.2)min in group A,significantly shorter than that in group B(36.1±7.6)min,submucosal injection number was(2.7±1.1)times in group A,significantly less than that in group B(3.5±1.2)times,submucosal dissection rate was(0.4±0.2)cm2/min in group A,significantly faster than that in group B(0.2±0.1)cm2/min,the differences were statistically significant(P<0.05).Conclusion Compared with traditional ESD,clip-line traction can provide a better surgical field and more effective dissection for large LST in rectum and sigmoid colon.

Objective To investigate the clinical efficacy of endoscopic pterygoid nerve combined with anterior ethmoidal nerve transection in treatment of allergic rhinitis with nasal polyps.Method A clinical practice study was conducted on 108 patients with allergic rhinitis complicated with nasal polyps.After enrollment,they were randomly divided into two groups,54 cases were included in the control group and treated with conventional endoscopic sinus surgery,while 54 cases were included in the observation group and treated with conventional endoscopic sinus surgery combined with pterygoid nerve and ethmoidal nerve transection.The nasal pain visual analogue scale(VAS),rhinoconjunctivitis quality of life questionnaire(RQLQ),asthma quality of life questionnaire(AQLQ),total effective rate,and complication rate of the two groups were compared before treatment and 1 week,2 weeks,3 weeks,3 months,6 months,and 1 year after treatment.Result Compared with the control group,the total effective rate of treatment was 98.15%in the observation group,which was higher than 85.19%in the control group,the difference was statistically significant(P<0.05).Before treatment,there were no significant differences in nasal resistance(NR),mucociliary transport rate(MTR),mucociliary transport time(MTT),and Lund-Kennedy scores between the two groups,the differences were no statistically significant(P>0.05).After treatment,MTR in two groups increased compared with before treatment,and the observation group was higher than the control group,the difference was statistically significant(P<0.05);NR,MTT,and Lund-Kennedy scores in two groups decreased,and the observation group was lower than the control group,the differences were statistically significant(P<0.05).Before treatment,there were no significant differences in nasal pain VAS,RQLQ,and AQLQ between the two groups,the differences were no statistically significant(P>0.05).After treatment,at 1 week,2 weeks,3 weeks,and 3 months,6 months,and 1 year,the observation group had lower nasal pain VAS compared to the control group,while the observation group had lower RQLQ and higher AQLQ compared to the control group,the differences were statistically significant(P<0.05).The incidence of complications in the observation group was lower than the control group,the difference was statistically significant(P<0.05).Conclusion The clinical efficacy of endoscopic pterygoid nerve combined with ethmoidal nerve transection for the treatment of allergic rhinitis complicated with nasal polyps is significant.It can not only effectively alleviate nasal pain in patients,but also improve their quality of life and reduce postoperative complications,which has clinical application value.

AIM To investigate the effects of Compound Dihuang Granules on biological function of dopaminergic neurons in rats with Parkinson's disease(PD)of Pattern of Yin-Deficiency with Stirring Wind.METHODS The PD animal model of Pattern of Yin-Deficiency with Stirring Wind established by intracerebral injection of 6-hydroxydopamine(6-OHDA)were randomly divided into the model group,the Midoba group(150 mg/kg)and the low,medium and high dose Compound Dihuang Granules groups(1.75,3.5 and 7 g/kg)for corresponding drug intervention,in contrast to the normal group,the sham operation group and the model group underwent 28-day normal saline administration.The rats had their general condition and neuroethology observed;their pathological changes of substantia nigra observed by HE staining;their mitochondrial structure of dopaminergic neurons in the damaged substantia nigra observed by transmission electron microscopy;and their expressions of DJ-1,IP3R,GRP75 and VDAC1 detected by immunohistochemistry,Western blot and RT-qPCR.RESULTS Compared with the normal group and sham operation group,the model group displayed increased rotational behavirors(P＜0.01),decreased swimming time score(P＜0.01),decreased hanging time(P＜0.01),decreased number of neurons in substantia nigra but more neurons with morphological damage,mitochondrial swelling and degeneration,mitochondrial crista disappearance,and decreased expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured side(P＜0.01).Compared with the model group,the Midoba group and the Compound Dihuang Granules groups demonstrated less rotational behavirors(P＜0.01),higher swimming time score(P＜0.01),longer hanging time(P＜0.01),less damage to mitochondrial morphology and structure,and higher expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured substantia nigra(P＜0.05,P＜0.01).The high-dose Compound Dihuang Granules presented equivalent efficacy to that of Midoba.CONCLUSION Compound Dihuang Granules may promote endoplasmic reticulum-mitochondria homeostasis,reduce mitochondrial damage and maintain the biological function of dopaminergic neurons by regulating the expressions of DJ-1,IP3R,GRP75 and VDAC1.

Objective To investigate the effects of arginine-glycine-aspartic acid(RGD)peptide hydrogels at differ-ent concentrations on the activation of Tenon capsule fibroblasts(HTF)and the expression of Yes-associated protein(YAP).Methods Three concentrations(0.5％,1.0％,2.0％)of RGD peptide hydrogels were prepared.Their internal microstructures were observed under a transmission electron microscope,and their elastic moduli(E)were measured using the rheological test.SD male rat models with conjunctival injury were established and divided into a blank group(no treatment),a low-concentration RGD group(subconjunctival injection of 0.5％RGD peptide hydrogel),a medium-con-centration RGD group(subconjunctival injection of 1.0％RGD peptide hydrogel),and a high-concentration RGD group(subconjunctival injection of 2.0％RGD peptide hydrogel).After one week,tissues were collected,and collagen fiber pro-liferation in the rat conjunctiva was observed after Masson staining.Immunohistochemistry was performed to detect the ex-pression of YAP and α-smooth muscle actin(α-SMA)in the rat conjunctiva.Scar formation cell models were constructed by stimulating HTF activation with transforming growth factor-β2(TGF-β2).According to experimental requirements,the cells were divided into a control group,a 0.5％RGD hydrogel group,a 1.0％RGD hydrogel group,a 2.0％RGD hydrogel group,and a normal group,and co-cultured for 24 hours.The relative expression levels of YAP,connective tissue growth factor(CTGF),α-SMA,fibronectin(FN),and type Ⅰ collagen(Col Ⅰ)proteins in HTF were detected by Western blot.The localization of cytoskeletal protein(F-actin)and YAP protein expression was observed under a confocal laser scanning mi-croscope.Results Electron microscopy showed that the crosslinking density of the nanofibers inside the RGD peptide hydrogel increased with the rise of the peptide concentration.Rheological measurements indicated that the E values of the 0.5％,1.0％,and 2.0％RGD peptide hydrogels were approximately 0.067 kPa,0.150 kPa,and 2.170 kPa,respectively.Masson staining results revealed that the area of collagen fiber proliferation in the conjunctiva of rats in the medium-concen-tration RGD group was significantly lower than that in the blank group(P＜0.05),while it showed no significant difference in the low-concentration and high-concentration RGD groups compared with the blank group(both P＞0.05).Immunohis-tochemistry results showed that the relative expression levels of YAP and α-SMA proteins in the conjunctiva of rats in the low-,medium-,and high-concentration RGD groups decreased compared to the blank group,and the decrease was the most significant in the medium-concentration RGD group(all P＜0.05).Cell immunofluorescence staining results indicated that the green fluorescence of F-actin was attenuated,and the red fluorescence of YAP was translocated from the nucleus to the cytoplasm in the 0.5％,1.0％,and 2.0％RGD hydrogel groups compared to the control group.Western blot results showed that the relative expression levels of YAP,CTGF,α-SMA,FN,and Col Ⅰ proteins in HTF were lower in the 0.5％,1.0％,and 2.0％RGD hydrogel groups compared to the control group,with the 1.0％RGD hydrogel group showing the most significant reduction(all P＜0.05).Conclusion With the optimal concentration of 1.0％,the RGD peptide hydro-gel can inhibit YAP protein expression and nuclear translocation and reduce HTF activation and the expression of fibrotic proteins,thus alleviating scar formation.

This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL^-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

The BTB (broad-complex, tramtrack, and bric-à-brac) domain is a highly conserved protein interaction motif in eukaryotes. They are widely involved in transcriptional regulation, protein degradation and other processes. Recently, an increasing number of studies have shown that these genes play important roles in plant growth and development, biotic and abiotic stress processes. Here, we summarize the advances of these proteins ubiquitination-mediated development and abiotic stress responses in plants based on the protein structure, which may facilitate the study of this type of gene in plants.
Eukaryota ; Plant Development/genetics* ; Proteolysis ; Ubiquitination

Objective To compare the therapeutic efficacy of whey protein powder in peritoneal dialysis hypoproteinemia patients.Methods From August 2019 to August 2020,100 patients with peritoneal dialysis hypoproteinemia from the Peritoneal Dialysis Center of the First Affiliated Hospital of Xi'an Jiaotong University were investigated.The patients were separated into 2 groups randomly;the study group were administered whey protein powder and the control group with high protein food.After 32 weeks of treatment,biochemical and biometric indices including hemoglobin(Hb),albumin(ALB),prealbumin(PA),triglyceride(TG),total cholesterol(TC),phosphorus(P),low density lipoprotein(LDL),high density lipoprotein(HDL),serum creatinine(Scr),blood urea nitrogen(BUN),estimated glo-merular filtration rate(eGFR),total spKt/Vurea(TKt/V),total creatinine clearance rate(TCcr),hand grip strength(HG),triceps skinfold(TSF),arm circumference(AMC),mid-arm muscle circumference(MAMC)were compared between groups.Results Compared with 0 week,at 16 and 32 weeks,ALB,PA,and HG were significantly increased in the study group(P＜0.05).Compared with the control group,ALB,PA,and HG increased significantly at 16 and 32 weeks in the study group(P＜0.05).There were no significant differences in TG,TC,HDL,LDL,eGFR,TKt/V,and TCcr at 0,8,16,and 32 weeks between the control and study groups(P＞0.05).Conclusion For patients with peritoneal dialysis hypoproteinemia caused by insufficient protein intake or excessive protein loss,the addition of whey protein during peritoneal dialysis can significantly improve the nutritional status of patients,with greater efficacy than a high protein diet alone.

Objective To explore the anti-atherosclerosis mechanism of Hirudo and its effect on autophagy in mice.Methods Forty healthy male ApoE-/-mice were randomly divided into model group,control group(3 × 10-3 g·kg-1·d-1 simvastatin)and experimental-L,experimental-M,experimental-H groups(0.45,0.9,1.8 g·kg-1· d-1,Maixuekang capsule).Eight healthy male C57BL/6J mice were divided into blank group.The mice were fed with common diet for 1 week.Then,except blank group,other groups were fed with high-fat diet.After 8 weeks of modeling,the atherosclerosis(AS)mice were given drugs orally once a day for 12 weeks,and fed with high-fat diet in the meantime.The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)in serum were determined by enzyme-linked immunosorbent assay(ELISA).The levels of Beclin-1,LC3 autophagy protein were detected by Western blot method.Results The IL-6 contents in the experimental-H,experimental-M,experimental-L,control,model and blank groups were(107.59±3.03),(99.31±5.12),(103.52±2.28),(98.68±4.68),(112.66±6.08),(93.98±3.43)pg·mL-1;the TNF-α contents were(538.41±30.26),(504.49±21.51),(538.51±19.05),(494.05±25.08),(578.53±26.32),(467.35±21.53)pg·mL-1.For the above indexes,the differences between model group and experimental-H group,experimental-M group,experimental-L group,control group,blank group were all statistically significant(all P＜0.05).The Beclin-1 protein expression levels in the experimental-H,experimental-M,experimental-L,control and model groups were 1.48±0.05,1.72±0.05,1.19±0.02,1.51±0.04,0.66±0.03;the LC3 Ⅱ protein expression levels were 1.53±0.01,1.83±0.02,1.16±0.01,1.90±0.01,0.49±0.01,and the differences between model group and experimental-H group,experimental-M group,experimental-L group,control group were all statistically significant(all P＜0.05).Conclusion Hirudo can significantly reduce the area of atherosclerotic plaque by regulating the level of autophagy.

Objective To investigate the protective effect of silymarin extract(SME)and its complex preparation on ethanol liver injury.Methods An ethanol liver injury model was established by gavage of 12 mL·kg-1 50％ethanol.Male mice were divided into blank group(distilled water),model group(ethanol liver injury model),SME-L,-H groups(6,20 mg·mL-1 SME),SME+Ganoderma lucidum extract(GLE)-L,-H groups(10,30 mg·mL-1 SME+GLE,SME∶GLE=1∶1),Jian An Shi Silymarin Pueraria Mirifica and Tansy tablets(JAS)-L,-H groups(68,204 mg·mL-1 JAS),there were 12 mice in each group.The serum levels of glutamic-oxaloacetic transaminase(GOT)in mice were measured by fully automated biochemical analyzer assay;the serum levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in mice were measured by enzyme-linked immunosorbent assay(ELASA);the hepatic tissue of oxidative stress indexes[catalase(CAT)and total superoxide dismutase(T-SOD)]were measured by ultraviolet spectrophotometer.Results The T-SOD activity in the blank group,model group,SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L and JAS-H groups were(192.54±49.00),(141.65±34.72),(205.83±32.77),(191.68±25.83),(192.31±28.79),(177.82±32.61),(218.58±74.80)and(210.24±31.65)U·mg·prot-1;CAT activity were(37.78±5.73),(28.92±8.44),(44.12±11.52),(41.41±9.15),(47.01±10.48),(41.63±8.95),(47.14±8.91)and(48.29±10.06)U·mg-1;GPT levels were(47.61±13.00),(97.84±26.00),(62.33±18.92),(51.84±17.91),(70.77±28.00),(58.00±21.27),(52.28±18.78)and(45.55±9.27)U·L-1;IL-6 levels were(21.03±1.52),(28.43±5.75),(21.90±3.24),(21.23±1.55),(22.26±2.58),(21.24±2.91),(22.17±4.14)and(21.14±3.02)pg·mL-1.Comparing the above indexes in the model group with the blank group,and comparing the above indexes in the SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L,JAS-H groups with the model group,the differences were statistically significant(all P＜0.01).The TNF-α levels in blank,model,SME-L,SME-H,SME+GLE-L,SME+GLE-H,JAS-L and JAS-H groups were were(28.07±7.72),(69.02±16.34),(40.29±8.94),(48.84±10.17),(41.91±14.96),(40.07±12.75),(50.72±11.44)and(45.05±11.34)pg·mL-1.Comparing the model group with the blank group,the SME,SME+GLE-L,-M and JAS,-M groups with the model group,the differences were statistically significant(all P＜0.01).Conclusion Silybum marianum extract and its compound preparation can increase the antioxidant level and reduce the inflammation of mouse liver,and have a certain improvement effect on liver injury caused by acute ethanol poisoning.