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.

This study was aimed to optimize the hydrolysis conditions of Coix prolamin, which occupied the highest content in total protein of it, and then evaluate the angiotensin-converting enzyme (ACE) inhibitory activity of the corresponding hydrolysates. Pepsin was used to hydrolyze prolamin and OPA method was applied to determine the degree of hydrolysis. The optimum conditions of hydrolyzing prolamin were obtained by orthogonal design subse-quently. The ACE inhibitory activity of small molecular weight (less than 3 kD) peptides gained by ultrafiltration of the hydrolysates was assayed by RP-HPLC method. The result showed that the optimized hydrolysis conditions were substrate concentration of 1%, enzyme-to-substrate ratio of 1:5 and hydrolysis duration for 48 h. The hydrolysates exhibited an ACE inhibitory ratio of (83.40 ± 0.93)% at the concentration of 0.1 mg·mL-1. It was concluded that prolamin hydrolysates displayed a high ACE inhibitory activity in vitro, which provided guidance for further research of antihypertensive component and development of functional food from Coix.

Objective To investigate the relationship and mechanism of the serum vitamin D levels with insulin sensitivity and adipokines APN,RBP4,visfatin and TNF-α in patients with gestational diabetes mellitus(GDM). Methods A total of 101 patients with gestational diabetes( GDM group) and 50 pregnant women with normal blood sugar( control group) were recruited to detect the serum levels of correlative biochemical indexes,APN,RBP4,visfatin,TNF-α and 25OHD3 using an HPLC method. Twenty-four patients in the GDM group diagnosed with vitamin D deficiency(<25 nmol·L-1 ) were randomized to receive either 1,25(OH) 2D3(0. 25 μg·d-1) orally or no treatment. Serum levels of correlative biochemical indexes were detected after 1 month. Results GDM patients had 25OHD3 levels significantly lower than the control group(P<0. 05),APN levels lower than the control group,and RBP4,visfatin,TNF-α levels significantly higher than the control group(all P<0. 05). After 1 month of 1,25(OH) 2D3 treatment,HOMA-IR increased,while HOMA-β decreased. APN levels was positively correlated(r= 0. 526) with 25OHD3;RBP4,visfatin,TNF-α decreased and were negatively correlated with 25OHD3( r values were -0. 272,-0. 153,-0. 072). Conclusion Vitamin D can reduce the adipokines RBP4,visfatin,TNF-α,increase APN,and thus play a protective role for gestational diabetes.

Objective:To investigate the use of the reference intervals for blood cell counting and the reference of industry standard in China.Methods:Information from all laboratories was collected using online questionnaire in 18 reference intervals survey in blood cell counting in 2019. The information includes the source of the reference intervals, the verification of the reference intervals, and the upper and lower limits of the reference intervals, the method used, the instrument, the reagent and the calibrator. Microsoft Excel 2007 software was used to analyze the results of all laboratories. The median and 95% confidence interval were calculated. The distribution of the reference intervals for blood cell counting and their conformance to industry standards were analyzed.Results:2, 869 labs reported the data. The main sources were industry standards and National Guide to Clinical Laboratory Procedures. The proportion was 33.30%-35.02% and 28.55%-30.90% respectively. 49.44%-55.13% of laboratories validated the reference interval when citing industry standards. The reference interval grouping of most laboratories (89.37%-91.69%) cited in RBC, Hgb and Hct were consistent with the industry standards. We compared the upper and lower limits of the reference intervals with that given by the industry standards, when the lower limit of the reference intervals of mean corpuscular hemoglobin concentration, absolute neutrophils count, absolute basophils count, absolute monocyte count, and lymphocyte percentage were compared. The upper limit of reference intervals of neutrophils percentage as well as upper and lower limits of reference intervals of mean corpuscular volume, mean corpuscular hemoglobin, absolute eosinophil count, basophils percentage, and monocyte percentage were also compared. The median and mode were equal and consistent with industry standards. For other labs, the upper and lower limits of the reference intervals were not consistent with the reference intervals given by the industry standards.Conclusion:The use of reference intervals for blood cell counting was not the same, and the implementation of industry standards was not optimistic. A considerable number of laboratories had not verified the reference intervals, so it was necessary to promote the industry standards for reference intervals.

Objective: To investigate the serotype and drug resistance of 815 Salmonella isolates from Hunan Province, so as to provide insights into management of Salmonella infections. Methods: Salmonella isolates were collected from stool samples of foodborne diarrheal patients and food samples in Hunan Province from 2020 to 2021, and serotyped. Antimicrobial susceptibility test was performed using the broth microdilution method. Results: A total of 10 groups and 39 serotypes were characterized in 815 Salmonella isolates. Among the 646 Salmonella isolates of human sources, 388 isolates were identified as serogroup B (60.06%), with S. typhimurium and its variants aspredominant serotypes (364 isolates, 56.35%), and among 169 foodborne isolates, 61 isolates were characterized as serogroup B (36.09%) with S. london as the predominant serotype (26 isolates, 15.38%). There were 597 antimicrobial resistant Salmonella isolates of human sources, with a drug resistance rate of 92.41%, and the percentage of ampicillin resistance was 81.58%. There were 140 foodborne antimicrobial resistant isolates, with a drug resistance rate of 82.84%, and the proportion of tetracycline resistance was 72.78%. However, Salmonella isolates from both humans and foods were sensitive to imipenem. In addition, there were 577 multidrug resistant Salmonella isolates, including 490 multidrug resistant isolates of human sources and 87 foodborne multidrug resistant isolates. Conclusions S. typhimurium and its variants and S. london were predominant serotypes of Salmonella isolates from 815 foodborne diarrheal patients and food samples in Hunan Province from 2020 to 2021, and a high rate of multidrug resistance was detected.

"Wise men could recognize similarities, but the fool only recognizes differences" in Su-wen, which expounded clinical thinking methods of Chinese medicine (CM). "To recognize similarities and differences simultaneously" is of important clinical significance in understanding the laws of diseases. CM pays much attention to recognize similarities, while modern medicine emphasizes the differences observed. In order to develop integrative medicine (IM), similarities recognition and differences identification must be combined together to innovate new thinking methods of IM.
Humans ; Integrative Medicine ; standards ; Medicine, Chinese Traditional

Background:TLR4 can mediate immune and inflammatory responses,TFF3,MUC2 are the intestinal mucosa protection factor and can maintain the intestinal barrier function. Aims:To investigate the effect of Anchang Yuyang decoction on colon tissue TFF3,MUC2 and TLR4 gene expressions in rats with ulcerative colitis. Methods:TNBS was used to establish ulcerative colitis model in rats. Ninety Wistar rats were randomly divided into normal control group,model group,low,moderate and high dose Anchang Yuyang decoction groups and mesalazine group,and distilled water,different concentrations of Anchang Yuyang decoction and mesalazine were given respectively. All the rats were sacrificed after 21 days. Colonic histopathological score was assessed,and RT-PCR was used to detect gene expressions of colon tissue TFF3, MUC2 and TLR4. Results:Compared with model group,histopathological score and TLR4 expression were significantly decreased in moderate,high dose Anchang Yuyang decoction groups and mesalazine group(P < 0. 05),expressions of TFF3 and MUC2 were significantly increased( P < 0. 05). Compared with moderate dose Anchang Yuyang decoction group,histopathological score in high dose Anchang Yuyang decoction group and mesalazine group was significantly decreased(P < 0. 05),and TFF3 expression was significantly increased( P < 0. 01). Compared with moderate dose Anchang Yuyang decoction group and mesalazine group,MUC2 expression in high dose Anchang Yuyang decoction group was significantly increased(P < 0. 01),and TLR4 expression was significantly decreased( P < 0. 01). Conclusions:Anchang Yuyang decoction can promote the repair of colonic mucosa in rats with ulcerative colitis,and its mechanism may be related to the increase of TFF3 and MUC2 gene expressions and down regulation of TLR4 gene expression.

Object: To study the proliferation of hCTGF on cells and its biological function on bone injury healing.Methods: The fibroblast with potential differentiation was transfected by eukaryotic gene delivery system and then transferred into the experimental animal model with bone fracture.The data were collected by molecular biological and clinical orthopedic technique detection analysis.Results: The results demonstrated an obvious proliferation of hCTGF on cells,suggesting that hCTGF have the biological activity of repairing bone injury via gene therapy.The results provide a new activity factor and treatment approach for bone injury in clinics.

Curcumin (Cur) is an important bioactive component of polyphenols in the rhizomes of Curcuma longa L., Tulipa gesneriana L. and other Curcuma plants. It has a wide range of pharmacological effects such as anti-tumor, anti-atherosclerosis, anti-inflammatory, and neuroprotection. Parkinson disease (PD) is a neurodegenerative disease that often occurs in the elderly. Its main pathological characteristics are the characteristic loss of substantia nigra dopaminer?gic neurons, the decrease of dopamine content in the striatum, and the formation of Lewy bodies. At present, the main methods of clinical treatment of PD include drug therapy and surgical operation, but due to its complicated pathogene?sis, they can only play a role in relieving, but cannot be completely cured. Modern pharmacological studies have shown that Cur has certain effects in the treatment of PD. ① Anti-oxidative stress: oxidative stress is closely related to the degeneration of dopaminergic neurons. Studies have found that Cur can increase the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), reduce malondialdehyde (MDA) content, thereby reducing oxidative stress damage and protecting dopaminergic neuron.②Reduce inflammation in brain tissue:neuroinflammation plays an impor?tant role in the development of PD. Reducing the level of inflammatory factors can have a certain therapeutic effect on PD. Studies have shown that high-dose Cur can reduce the levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis fac?tor-α (TNF-α) in brain tissue, reduce inflammation, inhibit further neuronal damage, improve learning and memory, and exert neuroprotective effects. ③ Activation of autophagy: the abnormal accumulation of α-Synuclein (α-Syn) in Lewy bodies is closely related to PD, and autophagy dysfunction leads to α-Syn clearance obstacles and an important factor of abnormal aggregation. Cur can increase the expression of microtubule-associated protein 1 light chain 3 (LC3-Ⅱ) and lysosome-associated membrane protein 2A (LAMP2A), and reduce the protein and mRNA expression of α-Syn. It can be seen that Cur promotes the elimination ofα-Syn and protects neurons from damage by activating autophagy.④Inhi?bition of mitochondrial dysfunction:mitochondria plays a central regulatory role in the process of cell apoptosis, and mito?chondrial dysfunction is related to reactive oxygen species, energy and mitochondrial membrane potential, which may cause substantia nigra striatal neuropathy. Experiments have shown that Cur can reduce the active oxygen content in PC12 cells induced by MPP+, maintain the normal membrane potential of mitochondria, thereby stabilizing mitochondrial function and inhibiting PC12 cell apoptosis. This study summarized the action mechanism of Cur in the treatment of PD, and clarified the basis of its pharmacodynamics, providing a reference for the clinical research and new drug develop?ment research of Cur in the treatment of PD.

Lyotropic liquid crystal system is formed by the amphiphilic molecules dissolving in polar solvents with a special geometric structure. Lamellar, cubic and hexagonal mesophases are some of the most common lyotropic liquid crystal systems. Recently, they have attracted much research attention because of their distinctive structures and physico-chemical properties(like strong bioadhesion, high permeability, low liquidity, and slow released drug), and have been widely used as carriers for drug delivery systems, especially in transdermal and mucosal fields. According to the research about lyotropic liquid crystal and nasal route of administration in our group, and the related references in recent years, we investigate the technical strategies about the using of lyotropic liquid crystal in transdermal and mucosal drug delivery system. Among them, we specially put the emphasis on the application prospects of lyotropic liquid crystal in the nasal mucosal administration, and then provide a theoretical basis and future research directions in the development of lyotropic liquid crystal in transdermal and mucosal administration fields.