1.Production and Identification of Broad Spectrum Monoclonal Antibody against a Group of Pyrethroid Insecticides
Mengtang WEN ; Yuan LIU ; Shuai YAN ; Xiao ZHANG ; Heng WANG ; Xianjin LIU
Chinese Journal of Analytical Chemistry 2014;(9):1245-1251
The objective of this study is to generate broad spectrum monoclonal antibody ( mAb ) against a group of pyrethroid insecticides and to identify its immunological characteristics. The generic hapten 3-phenoxy-benzoic acid ( PBA) was conjugated to carrier protein BSA by activated ester method. Balb/c mice were immunized with PBA-BSA. The titer of polyclonal antibody ( pAb ) was detected by indirect enzyme-linked immunosorbent assay ( ELISA) after five times immunization. The mouse with high titer and sensitivity was selected for cell fusing. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by indirect non-competitive ELISA based on the coating antigen PBA-ovoalbumin ( PBA-OVA ) . High-sensitivity and high-specificity mAb was prepared after subcloning using limiting dilution method. Purified mAb was obtained after purified by saturated ammonia sulfate precipitation and protein G affinity column. The immunological characteristics of mAb such as titer, antibody subtypes, affinity constant and the sensitivity to pyrethroid insecticides were characterized by indirect ELISA; The results of UV spectroscopy and SDS-PAGE showed that PBA-BSA artificial antigen was synthesized successfully. A hybridoma cell line (4H11) secreting anti-pyrethroid mAb was established. The titre of ascites was up to 1:6. 5×106, and the mAb was IgG1 subtype. The affinity constant of the mAb to PBA was about 2. 5×107 L/mol, with a IC50 value of 208. 83 μg/L and a detection limit of 21. 23 μg/L to PBA. Simultaneously, beta-cypermethrin, flucythrinate, cypermethrin and fenvalerate were sensitively recognized by the mAb with the IC50 of 1. 01, 2. 15, 3. 16 and 3. 67μg/L, respectively.
2.Development of peptidic MERS-CoV entry inhibitors.
Shuai XIA ; Qian WANG ; Shu-wen LIU ; Lu LU ; Shi-bo JIANG
Acta Pharmaceutica Sinica 2015;50(12):1513-1519
In 2012, a new SARS-like coronavirus emerged in the Middle East, namely the Middle East respiratory syndrome coronavirus (MERS-CoV). It has caused outbreaks with high mortality. During infection of target cell, MERS-CoV S protein S1 subunit binds to the cellular receptor (DPP4), and its S2 subunit HR1 and HR2 regions intact with each other to form a stable six-helix bundle to mediate the fusion between virus and target cell membranes. Hence, blocking the process of six-helix bundle formation can effectively inhibit MERS-CoV entry into the target cells. This review focuses on the recent advance in the development of peptidic entry inhibitors targeting the MERS-CoV S2 subunit.
Antiviral Agents
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pharmacology
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Coronavirus Infections
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drug therapy
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Dipeptidyl Peptidase 4
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metabolism
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Drug Design
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Humans
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Middle East Respiratory Syndrome Coronavirus
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drug effects
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physiology
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Peptides
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pharmacology
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Spike Glycoprotein, Coronavirus
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metabolism
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Virus Internalization
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drug effects
3.Preoperative management of cardiac surgery with glucose-6-phosphate dehydrogenase deficiency
Hai-yong, WANG ; Yi-yao, JIANG ; Wen-bin, ZHANG ; Jian-fei, SONG ; Shuai-zhou, LIU
Chinese Journal of Endemiology 2011;30(6):691-693
Objective To observe the perioperative management of cardiac surgery and extracorporeal circulation method in patients with glucose-6-phosphate dehydrogenase deficiency(G6PD).Methods Ten patients with G6PD deficiency underwent uneventful cardiac surgery procedures between January 2005 and December 2010.Twenty patients who had non-G6PD deficiency were as a control group,the selected conditions were the same gender,age,body mass,the risk of heart disease surgery.The preoperative management in patients with G6PD deficiency mainly focused on avoiding the drugs implicated in haemolysis,reducing the surgical stress,using moderate hypothermia extracorporeal circulation and enhancing blood conservation.Observed indicators included the assisted ventilation time,urine volume,the drainage volume of chest tube,the amount transfusion of red blood cells and plasma,the level of hemoglobin and serum total bilirubin in the 2nd day after surgery,ICU stay.Results Compared with the control group,patients with G6PD deficiency had no significant difference in duration of ventilation after the operation,drainage,urine,Hgb,bilirubin levels,and blood transfusion[(9.3 ± 4.5)h vs (8.6 ± 5.7)h,(2100 ±670)ml vs (1950 ± 490) ml,(253 ± 146)ml vs (260 ± 120)ml,(1.3 ± 1.0)U vs (1.8 ± 1.2)U,(96 ± 25)g/L vs (99 ± 12)g/L,and (24 ± 8)μmol/L vs (27 ± 1 l)μmol/L,t =0.978,2.032,1.257,0.891,2.182,2.271,and 1.329,all P > 0.05].The duration of ICU discharge was significantly longer in the glucose-6-phosphate dehydrogenase deficient group[ (2.6 ± 0.6)d vs (1.8 ± 1.5)d,t =2.704,P < 0.05].Conclusions Cardiac surgery can be performed safely in patients with G6PD deficiency with enhanced perioperative management.
4.Effect of emodin on the levels of hypoxia inducible factor-1 α and glycogen synthase kinase-3β in rats with severe acute pancreatitis
The Chinese Journal of Clinical Pharmacology 2017;33(14):1332-1335
Alstract:Objective To study the effect of emodin on the levels of hypoxia inducible factor-1 α (HIF-1 α) and glycogen synthase kinase-3β (GSK-3β) in rats with severe acute pancreatitis (SAP).Methods A total of 60 rats were randomly divided into blank group,sham-operation group,model group,control group and test group,each group had 12 rats.Model and test groups were injected with sodium taurocholate 0.1 mL / 100 g to establish rat model of severe pancreatitis with renal injury.Sham-operation and control groups were gently flipped the duodenum and pancreas after opening the abdomen.Blank group was not given any treatment.Blank,sham-operation and model groups were given 5 μL · g-1 0.9% NaCl q6 h,intraperitoneal injection.Control and test groups were given 5 g · L-1 emodin with the dose was 25 μg · g-1 q6 h,intraperitoneal injection.The serum creatinine (SCr),urea nitrogen (BUN),HIF-lαt and GSK-3β were compared between the five groups after operation 3,6 and 12 h.Results The pancreatic histopathology score in model group was (9.84 ± 1.05) points,which in blank,sham-operation,control and test groups were (1.30 ± 0.14),(1.39 ± 0.14),(1.40 ± 0.15),(2.73 ± 0.30) points,the differences were significant (P < 0.05),and there were significant difference between test group and blank,sham-operation,control groups (P < 0.05).After operation 3 h,the main indexes in blank,sham-operation,control,model,test groups were compared,SCr were (60.46 ± 6.78),(61.52 ± 6.78),(60.38 ± 6.74),(120.57 ± 13.43),(95.35 ± 9.85) μmol· L-1;BUN were(7.47 ±0.79),(8.02 ±0.92),(7.93 ±0.83),(12.49 ± 1.53),(8.56 ±0.89) mmol· L-1;HIF-1α were (225.46 ±23.57),(210.57 ±23.58),(229.67 ±6.74),(160.46 ±17.47),(144.57±14.85)μg· L-1;GSK-3β were (6.59±0.69),(6.57 ±0.69),(6.74±0.68),(19.95 ±2.13),(10.56 ± 1.39) μg · L-1.After operation 6 h,the main indexes in blank,sham-operation,control,model,test groups were compared,SCr were (60.57 ±6.79),(60.55 ±6.76),(59.50 ±6.76),(143.57 ± 15.47),(110.57 ±12.55)μmol· L-1;BUN were (7.65 ±0.84),(8.11 ±0.93),(8.29±0.92),(15.46±1.64),(12.35 ± 1.37)mmol · L-1;HIF-1α were (226.46 ±24.04),(222.46 ±23.57),(230.57 ±24.05),(155.36 ± 15.74) (127.57 ± 12.84) μg · L-1GSK-3β were (6.25 ±0.67),(6.71 ±0.69),(6.82 ±0.71),(25.34 ±2.64),(11.56 ± 1.27)μg · L-1.After operation 12 h,the main indexes in blank,sham-operation,control,model,test groups were compared,SCr were (60.61 ± 6.79),(61.68 ± 6.79),(61.54 ± 6.78),(166.45 ±17.05),(131.45 ±13.46) μmol · L-1;BUN were (8.03 ±0.85),(7.98 ±0.82),(8.79 ±0.93),(18.66±19.46) (15.35 ± 1.63)mmol · L-1-,HIF-1α were (219.57 ±23.63),(226.35 ±24.04),(220.25 ±24.04) (148.46 ± 18.94) (117.46 ± 12.04) μg · L-1,,GSK-3β were (6.77 ± 0.69),(6.70 ±0.70),(6.86 ±0.70),(35.02 ± 3.76),(20.35 ± 2.13) μg · L-1.After operation 3,6 and 12 h,the SCr,BUN,HIF-1 α and GSK-3β in blank,sham-operation and control groups were significantly different with those of model group (P < 0.05),and those indexes were significantly different between test and model groups after operation 3,6 and 12 h (P < 0.05).Conclusion Emodin may enhance the ability of renal cells to hypoxia by increasing the level of serum HIF-1α and GSK-3β,thus playing a protective role in rats of severe acute pancreatitis with renal injury.
5.The synergistic effect of SARS-CoV-2 spike protein, angiotensin-converting enzyme 2 and protease on corona virus disease 2019
Yue ZHAO ; Wen-bin LIU ; Zi-shuai LI ; Guang-wen CAO
Shanghai Journal of Preventive Medicine 2021;33(3):237-
The severe acute syndrome coronavirus 2(SARS-CoV-2)is responsible for the global pandemic of 2019 novel coronavirus disease(COVID-19). Understanding the pathogenic mechanisms of SARS-CoV-2 is of great significance to the prevention and control of the COVID-19 epidemic. The spike(S)protein of SARS-CoV-2 is a major target of protective immunity. The mutations of S protein impact the transmission and pathogenic capacity of SARS-CoV-2. In addition, angiotensin-converting enzyme 2(ACE2)is the receptor of SARS-CoV-2 S protein and mediates viral entry into target cells. The genetic polymorphisms of the ACE2 gene may lead to the difference of incidence of COVID-19 among different populations. Protease can promote the infection of SARS-Cov-2 by activating the S protein. This review summarized the current understanding of the function and genetic variants of S protein, ACE2, and the related proteases. The synergistic effect of these three molecules on the susceptibility of SARS-CoV-2 infection was further discussed.
6.Correlation between measurement error of oxygen concentration and service time of ventilator's oxygen cell
Can DONG ; Shuai LIU ; jun Wan SHUAI ; zhe Wen LI ; Yong CHAO
Chinese Medical Equipment Journal 2017;38(11):83-85
Objective To analyze the relationship between the measurement error of oxygen concentration and service time of oxygen cell of the ventilator. Methods The present situation of the ventilator was analyzed. Some cases of alarms for oxygen concentration failures were taken in statistical analysis on 3-a quality control detection results of the ventilators in some hospital. Results There was an approximately linear correlation between the measurement error of oxygen concentration and service time of oxygen cell, and the measurement error of oxygen concentration was increased significantly 430 days after the oxygen cell enabled. Conclusion The measurement error of oxygen concentration quality control could be used as a reference for performance evaluation and replacement of oxygen cell.
7.Simultaneous determination of alizarin and rubimaillin in Rubia cordifolia by ultrasound-assisted ionic liquid-reversed phase liquid chromatography.
Hong-shuai YANG ; Ju WANG ; Cui GUO ; Wei LIU ; Yuan-yuan CHEN ; Jin-feng WEI ; Wen-yi KANG
China Journal of Chinese Materia Medica 2015;40(13):2617-2623
Four kinds of ionic liquids were adopted to analyze the content of rubimaillin and alizarin in Rubia cordifolia roots with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 microm), was used. Methanol and 0.4% acetic acid-water as mobile phase with flow rate at 0.85 mL min(-1), gradient elution, detection wavelength at 250 nm, chromatographic column temperature was controlled at room temperature. The result showed that rubimaillin and alizarin had the highest extraction yield when the [ HMIM] PF6methanol solution concentration of 0.6 mol x L(-1) as extraction solvent and the conditions were solid-liquid ratio of 1:80 (g x mL(-1)). Under the optimal extraction conditions, the content of alizarin from 0.01 to 0.04 microg showed a good linearity (r = 0.9999), the average recovery was 97.12%, the content of rubimaillin from 0.41 to 1.35 microg showed a good linearity (r = 0.9999), the average recovery was 98.10%. This experiment adopted environmentally friendly reagent as extraction solvent, the extraction efficiency was improved, and the environmental pollution caused by organic solvent was avoided, the harm of human body aslo was reduced. This method was simple and reliable, its repeatability was also very good, which had an important significance in the study of traditional Chinese medicine active ingredient extraction methods.
Anthraquinones
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analysis
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Chromatography, Reverse-Phase
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methods
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Ionic Liquids
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chemistry
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Pyrans
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analysis
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Rubia
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chemistry
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Ultrasonics
8.Determination of oleanic acid and paeoniflorin in Paeonia lactiflora by ultrasound-assisted ionic liquid-reversed phase liquid chromatography.
Wei LIU ; Dong-dong LI ; Hong-shuai YANG ; Yuan-yuan CHEN ; Jin-feng WEI ; Wen-yi KANG ; Xiu-chun GUO
China Journal of Chinese Materia Medica 2015;40(3):443-449
Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.
Chromatography, Reverse-Phase
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methods
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Glucosides
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analysis
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Ionic Liquids
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chemistry
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Monoterpenes
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analysis
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Oleanolic Acid
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analysis
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Paeonia
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chemistry
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Ultrasonics
9.Stress distribution on the maxilla when wearing the Twin-block appliance for Class Ⅱ malocclusion
Shuai LI ; Hua LIU ; Yonghui SHANG ; Yicong LIU ; Qihang ZHAO ; Wen LIU
Chinese Journal of Tissue Engineering Research 2025;29(5):881-887
BACKGROUND:The Twin-block orthodontic appliance is commonly used for the correction of Class Ⅱ malocclusion.Its mechanism of action in stimulating mandibular growth has been confirmed in many studies,but its impact on maxillary growth is not very clear. OBJECTIVE:By establishing a finite element model to analyze the stress distribution of the maxillary complex,surrounding bone sutures,and maxillary dentition in patients with Class Ⅱ malocclusion wearing Twin-block orthodontic appliances. METHODS:One patient with Class Ⅱ malocclusion who underwent orthodontic treatment at Qingdao Hospital/Qingdao Municipal Hospital of Shandong Rehabilitation University was selected.The bite force data of the patient when wearing the Twin-block orthodontic appliance was measured,and CBCT data were collected.A finite element model was established,including the maxillary complex,peripheral sutures,Twin-block orthodontic appliance,and maxillary dentition.ABAQUS software was used to simulate the stress distribution in the maxilla and maxillary dentition when the patient was wearing the Twin-block appliance. RESULTS AND CONCLUSION:The equivalent stress on the maxillary anterior teeth was significantly smaller than that on the posterior teeth,and the maximum equivalent stress on both sides of the teeth were 4.797 5 Mpa and 8.716 1 Mpa,respectively,which were located at the first premolar.The maximum displacements were presented at the maxillary incisors on both sides of the teeth,which were 0.080 5 mm and 0.081 0 mm,respectively.The maximum equivalent stress on the bone suture was 1.284 Mpa,which was mainly concentrated in the pterygopalatine suture and the frontal-maxillary suture on both sides,and there was almost no difference in the force of the rest of bone sutures;the maximum displacement of the bone suture was 0.07 mm,with the pterygopalatine suture having the largest displacement,followed by the frontal-maxillary suture.The maximal equivalent stress on the maxillary complex was 27.18 Mpa,which was mainly concentrated on both sides of the anterior pyriform foramen of the maxilla,around the nasofrontal suture and around the pterygopalatine suture at the posterior part of the jaws.The maximal displacement of the maxilla was 0.07 mm,which was mainly concentrated on the maxillary alveolar bone.All these findings show that the occlusal force acts on the maxillary complex through the Twin-block appliance,resulting in clockwise rotation of the maxilla and steepening of the dentition plane.Measures should be taken to compensate for this tendency,for example,by considering maxillary molar elongation and intrusion in the process of occlusion,which are not only able to flatten the occlusal plane,but facilitate the mandibular protraction,thereby further improving Class Ⅱ malocclusion orthodontic treatment.
10.Three-dimensional finite element study on the effect of posterior tooth forward movement on temporomandibular joint stress in orthodontic reduction patients
Yonghui SHANG ; Shuai LI ; Yicong LIU ; Qihang ZHAO ; Wen LIU
Chinese Journal of Tissue Engineering Research 2024;28(34):5516-5520
BACKGROUND:Temporomandibular joint disorders are closely related to high stress in temporomandibular joint.With the change of molar position after tooth reduction extraction,the establishment of new occlusal relationship often leads to the change of internal stress environment of the temporomandibular joint. OBJECTIVE:To analyze the stress distribution of temporomandibular joint in patients undergoing orthodontic reduction tooth extraction with different degrees of molar forward movement using the three-dimensional finite element model of the maxillary complex and temporomandibular joint. METHODS:A case of individual normal occlusal patient was selected from the Orthodontics Department of Qingdao Municipal Hospital,Shandong Province,and the finite element models of 1/3 anterior molar space(extraction of four second premolar teeth)before and after reduction and 2/3 anterior molar space(extraction of 4 second premolar teeth)after reduction were established based on the cone-beam CT and MRI data.ABAQUS software was used to analyze the stress distribution of various parts of the temporomandibular joint during the interposition of tooth tips. RESULTS AND CONCLUSION:The stress distribution of the condyle,articular disc,and osteoarticular fossa in the model before and after the reduction was basically the same.The stress of the condyle was mainly distributed in the anterior and apical part of the condyle,the stress of the articular disc was mainly distributed in the middle band and lateral part of the articular disc,and the stress of the articular fossa was mainly concentrated in the anterior and apical part of the articular fossa.However,the equivalent stress value of the condyle,articular disc and articular fossa decreased after reduction.After orthodontic reduction extraction,the equivalent stress values of condyle and articular disc in the 1/3 anterior molar space model were smaller than those in the 2/3 anterior molar space model.From the perspective of biomechanics,orthodontic reduction extraction can reduce the stress of the temporomandibular joint and provide a good biomechanical environment.