Objective Evaluate the effects of different solvent extracts from Jujing Pills on improving retinal damage in kidney Yang deficiency aging mice and explore the underlying mechanism.Methods A composite modeling method of subcutaneous injection of D-galactose and hind limb injection of hydrocortisone was used to establish a kidney Yang deficiency type aging mouse model,and various solvent extract of Jujing Pills were given for intervention treatment.The improvement effect of different solvent extracts of Jujing Pills on aging mice with kidney Yang deficiency was evaluated by observing physical signs,measuring serum oxidative stress marker levels,and cyclic nucleotide levels.Optical coherence tomography(OCT)was used to detect the condition of the mouse retina,HE staining was used to detect the degree of retinal cell damage,TUNEL staining was used to detect retinal cell apoptosis,assay kit was used to detect oxidative stress factor expression,enzyme-linked immunosorbent assay was used to detect the expression of ciliary neurotrophic factor(CNTF),brain-derived neurotrophic factor(BDNF),and nerve growth factor(NGF)in the mouse eyeball,and Western blotting(WB)was used to quantitatively analyze the expression levels of apoptosis and neurotrophic pathway related proteins in the mouse retina tissue.Results Compared with the control group,the model group mice showed slow weight gain,reduced activity,decreased expression of antioxidant factors such as superoxide dismutase(SOD)and glutathione(GSH)in serum(P<0.01),and decreased cAMP/cGMP ratio(P<0.01).Different solvent extracts of Jujing Pills significantly increased the expression of antioxidant factors such as SOD and GSH(P<0.05),reduced the production of oxidative product malondialdehyde(MDA)(P<0.01),and increased the cAMP/cGMP ratio(P<0.01);Significantly increased the retinal thickness of model mice(P<0.01),improved retinal damage and inhibited retinal cell apoptosis(P<0.01),and significantly downregulated the ratio expression levels of Cleaved Caspase-3 and Caspase-3(P<0.01);The expression of CNTF,BDNF,and NGF was upregulated by water extracted medicinal residue alcohol extract,water extracted alcohol sediment,and total extract(P<0.01).The phosphorylation levels of CREB and ERK in the eyeball were significantly upregulated by water extracted medicinal residue alcohol extract and total extract(P<0.05).Conclusion Different solvent extracts of Jujing Pills can exert different pharmacological effects on improving retinal damage in aging mice with kidney yang deficiency model.Its mechanism of action is related to improving retinal apoptosis caused by oxidative stress and increasing the expression of nutritional factors.

Objective Evaluate the effects of different solvent extracts from Jujing Pills on improving retinal damage in kidney Yang deficiency aging mice and explore the underlying mechanism.Methods A composite modeling method of subcutaneous injection of D-galactose and hind limb injection of hydrocortisone was used to establish a kidney Yang deficiency type aging mouse model,and various solvent extract of Jujing Pills were given for intervention treatment.The improvement effect of different solvent extracts of Jujing Pills on aging mice with kidney Yang deficiency was evaluated by observing physical signs,measuring serum oxidative stress marker levels,and cyclic nucleotide levels.Optical coherence tomography(OCT)was used to detect the condition of the mouse retina,HE staining was used to detect the degree of retinal cell damage,TUNEL staining was used to detect retinal cell apoptosis,assay kit was used to detect oxidative stress factor expression,enzyme-linked immunosorbent assay was used to detect the expression of ciliary neurotrophic factor(CNTF),brain-derived neurotrophic factor(BDNF),and nerve growth factor(NGF)in the mouse eyeball,and Western blotting(WB)was used to quantitatively analyze the expression levels of apoptosis and neurotrophic pathway related proteins in the mouse retina tissue.Results Compared with the control group,the model group mice showed slow weight gain,reduced activity,decreased expression of antioxidant factors such as superoxide dismutase(SOD)and glutathione(GSH)in serum(P<0.01),and decreased cAMP/cGMP ratio(P<0.01).Different solvent extracts of Jujing Pills significantly increased the expression of antioxidant factors such as SOD and GSH(P<0.05),reduced the production of oxidative product malondialdehyde(MDA)(P<0.01),and increased the cAMP/cGMP ratio(P<0.01);Significantly increased the retinal thickness of model mice(P<0.01),improved retinal damage and inhibited retinal cell apoptosis(P<0.01),and significantly downregulated the ratio expression levels of Cleaved Caspase-3 and Caspase-3(P<0.01);The expression of CNTF,BDNF,and NGF was upregulated by water extracted medicinal residue alcohol extract,water extracted alcohol sediment,and total extract(P<0.01).The phosphorylation levels of CREB and ERK in the eyeball were significantly upregulated by water extracted medicinal residue alcohol extract and total extract(P<0.05).Conclusion Different solvent extracts of Jujing Pills can exert different pharmacological effects on improving retinal damage in aging mice with kidney yang deficiency model.Its mechanism of action is related to improving retinal apoptosis caused by oxidative stress and increasing the expression of nutritional factors.

Age-related macular degeneration(AMD)is a serious threat to the visual health of the elderly,and the dysfunction of retinal pigment epithelial cells(RPE)is a significant etiology risk.Aging process leads to RPE repli-cation senescence,and some environment factors like light exposure and cigarette exposure may lead to RPE stress premature aging,and the decreased lysosomal digestion ability of senescent RPE cells may lead to the accumulation of lipofuscin,triggering the occurrence of early AMD.A series of homeostatic imbalances in aging retina,such as cell senescence-renewal imbalance,oxidative stress-antioxidant imbalance,chronic inflammatory-anti-inflammatory imbalance,intestinal barrier and intestinal microbiota imbalance and pro-angiogenesis-antiangiogenic imbalance all contribute to the development of AMD.

Objective To study the effective fraction and mechanism of Lycium barbarum leaves on improving learning and memory ability of subacute aging mice induced by D-galactose injection.Methods The model of subacute aging mice was developed by injection of D-galactose subcutaneously,and different extracts of Lycium barbarum leaves were prepared.The effects of the extracts of Lycium barbarum leaves on the learning and memory ability of model mice were evaluated by Y maze experiment and new object recognition experiment.The pathomorphological changes of hippocampus in mice were observed by hematoxylin-eosin and Nissl staining.The levels of brain-derived neurotrophic factor(BDNF),nerve growth factor(NGF),glial cell line-derived neurotrophic factor(GDNF),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interferon-γ(IFN-γ)and interleukin-10(IL-10)in hippocampus of mice were detected by enzyme-linked immunosorbent assay.The activities of superoxide dismutase(SOD)and the contents of glutathione(GSH)and malondialdehyde(MDA)in hippocampus of mice were detected by related assay kits.Detection of apoptosis in the hippocampal region of mouse brain tissue using the TUNEL method.Western blotting analysis was used to detect the expressions of antioxidant proteins Nrf2,HO-1 and apoptotic proteins Caspase-3,Caspase-9 in hippocampus of mice.Results The water extraction part and 80%alcohol precipitation supernatant part of Lycium barbarum leaves significantly improved the learning and memory ability of model mice,improved the pathological damage of hippocampus in mice,increased the number of Nissl bodies in hippocampus of mice,and promoted the expression of neurotrophic factors BDNF,NGF and GDNF,and promoted the expression of neurotrophic factors BDNF,NGF and GDNF.Pro-inflammatory factors TNF-α,IL-1β and IFN-γ expression declines while anti-inflammatory factor IL-10 expression rises.The activity of SOD and the expression of GSH were increased,and the expression of MDA was decreased.Increase the expression of Nrf2 and HO-1 antioxidant proteins;reduce the expression of Caspase-3 and Caspase-9 apoptosis pathway proteins.Inhibition of apoptosis in the hippocampal region of mouse brain tissue using a model.Conclusion The water extracts and 80%alcohol precipitation supernatant extracts of Lycium barbarum leaves are the effective fractions of Lycium barbarum leaves to improve the learning and memory ability of D-galactose-induced subacute aging mice,and its mechanism might be related to the inhibition of neuronal apoptosis caused by oxidative stress and inflammation.

OBJECTIVE To optimize the deproteinization process of Isatidis Radix polysaccharides and further explore its immu-nomodulatory activity,and to provide a scientific basis for the development and utilization of it.METHODS The optimum conditions of enzymatic deproteinization were optimized by a single factor combined with the Box-Behnken response surface method.The chemical composition and structural characteristics of deproteinized Isatidis Radix polysaccharides were analyzed by UV-visible spectrum,Fourier transform-infrared spectroscopy,high-performance gel permeation chromatography,high-performance liquid chromatography and scan-ning electron microscopy.The effects of deproteinized Isatidis Radix Polysaccharide on neutrophils,macrophages,IL-1β and IL-6 in zebrafish were investigated by using a zebrafish immunocompromised model.RESULTS The optimal enzymatic deproteinization process was as follows:trypsin 500 U·mL-1,pH 8.0,enzymatic hydrolysis time 5 h,enzymatic hydrolysis temperature 37℃.The deproteinization rate was(86.39±0.07)%,and the comprehensive score was(91.15±0.37)%.Ultraviolet,infrared spectroscopy scanning and scanning electron microscopy showed that the protein contained in the crude polysaccharide could be removed by enzymat-ic method.The relative molecular weight of the polysaccharides were between 5.82 and 60.26 kDa.The monosaccharide mole compo-sition was mannose ∶ rhamnose ∶ galacturonic acid ∶ glucose ∶ galactose ∶ arabinose=2.17 ∶ 0.96 ∶ 2.90 ∶ 83.25 ∶ 4.88 ∶ 5.84.The results of immune activity evaluation showed that when the concentration of deproteinized Radix Isatidis polysaccharides was 50～300 μg·mL-1,it could significantly increase the density of zebrafish immune cells,increase the number of macrophages,and reduce the content of IL-1β and IL-6 in immunocompromised zebrafish,thus exerting immunomodulatory effects.CONCLUAION The enzy-matic method can effectively remove the proteins contained in the crude polysaccharides of Isatidis Radix,and the deproteinized Isatidis Radix polysaccharides have certain immunomodulatory effects.

OBJECTIVE To investigate the effect of Lycii Fructus-Chrysanthemi Flos herbal pair on the occurrence and progres-sion of light source-induced myopia in guinea pigs.METHODS The three-band fluorescent lamp was used to irradiate guinea pigs to establish a model of light source-induced myopia.Lycii Fructus-Chrysanthemi Flos herbal pair extract was added in the feed to raise guinea pigs.Ophthalmic A-type ultrasonic measuring instrument was used to detect the eye axis.HE staining was used to observe pathological damage to the retina of the guinea pigs.Enzyme-linked immunosorbent assay(ELISA)was used to detect dopamine(DA)and melatonin(MT)levels in guinea pig retinas,and transcriptomic analysis and other methods were used to explore the effects of Lycii Fructus-Chrysanthemi Flos on light-induced myopia in guinea pigs.RESULTS After 6 weeks,compared with the normal group,the eye axis of model group guinea pigs increased significantly(P<0.01),thus,the myopia model was successfully established.Lycii Fruc-tus-Chrysanthemi Flos herbal pair extract could effectively slow down the thickening of the lens and the deepening of the vitreous cavi-ty,thereby delaying the overgrowth of the eye axis in model guinea pigs.The retinal tissue of guinea pigs in the model group became thinner,the thickness and number of cells in the inner and outer nuclear layer of the retina were significantly reduced,the ganglion nu-clei were sparse,the vacuolization of the ganglion cell layer was obvious.Lycii Fructus-Chrysanthemi Flos extract could protect retinal cells.The thickness and cell number of the retinal inner nuclear layer and outer nuclear layer were increased,and ganglion cell nuclei were increased.ELISA assay showed that the concentrations of DA and MT in the retina of model group guinea pigs were significantly reduced(P<0.05),and the medium and high doses of Lycii Fructus-Chrysanthemi Flos herbal pair extract could effectively increase the concentration of dopamine in the retina and serum of guinea pigs in the model group(P<0.01,P<0.001).The differential genes screened by eye transcriptomics analysis were mainly enriched in PPAR signals pathway,collagen-containing extracellular matrix and other related pathways.ELISA results showed that the high dose of Lycii Fructus-Chrysanthemi Flos extract could significantly increase the content of PPARγ and COL1A1(P<0.01)and reduce the content of SMA(P<0.05).CONCLUSION The Lycii Fructus-Chry-santhemi Flos herbal pair extract has effects on delaying the occurrence and progression of light source-induced myopia in guinea pigs,protecting retinal cells,improving DA secretion disorders caused by artificial light environment,and regulating the level of PPAR,which has positive significance for the prevention and control of light source-induced myopia and provides a scientific basis for the ra-tional use of drugs in clinical practice and the creation of eye health products.

The concept,connotation and extension,goals and tasks of the discipline of Chinese medicinal materials resource chem-istry have been proposed and developed for 20 years.Looking back at the 20-year construction and development process,continuous exploration and innovative practice have been carried out around the scientific production and effective utilization of traditional Chinese medicinal materials.The theoretical connotation has been further enriched,the research mode has been further improved,and the tech-nical system has been further expanded.A series of research results have been formed and promoted for application,serving the high-quality development of the traditional Chinese medicinal materials industry,and contributing to the improvement of quality,efficiency,and green development of the entire industry chain of Chinese medicinal resources.However,with the rapid growth of Chinese medici-nal materials industry and the continuous expansion and extension of the industry chain,the waste and by-products generated in the production process of Chinese medicinal agriculture and industry are increasing day by day,causing resource waste and environmental pollution,which has become a new major problem facing the development of the industry.This article focuses on the establishment and case analysis of a model for the full industry chain recycling and low-carbon green development of Chinese medicinal materials,as well as the creation of an ecological industry demonstration park for the recycling of Chinese medicinal materials.It showcases the phased a-chievements made in recent years,aiming to provide demonstration and reference for the low-carbon and green transformation of the Chinese medicinal materials industry from a linear economy model to a circular economy model.It provides reference for improving the efficiency of Chinese medicinal materials utilization and creating new quality productivity,and helps promote low-carbon and green de-velopment in the field of Chinese medicinal materials industry.

OBJECTIVE: To establish a cell lines for quality control of prenatal genetic diagnosis. METHODS: The recombined SV40LTag-pcDNA3.1(-) vector was constructed and transfected by lipidosome into human amniotic fluid cells with common aneuploidy. Positive clones were screened by G418, and the immortality of transfected cell line was identified. RESULTS: Cell line with karyotype of 46, XY, t(8;19)(q24.3;q13.1) from primary amniotic fluid cells was established. Karyotype analytical results indicated that the cell line at its 15th generation maintained the same karyotype of its primary cell. CONCLUSIONS Gene can lead to immortality of amniotic fluid cells, which contributes to preparing cell lines for internal and external quality control in prenatal genetic diagnosis.
Antigens, Polyomavirus Transforming ; genetics ; Cell Line ; Female ; Genetic Vectors ; Humans ; Karyotype ; Pregnancy ; Prenatal Diagnosis ; methods ; Quality Control ; Recombinant Proteins ; genetics ; Transfection

To investigate the protective effect of the seed oil of Abelmoschus esculentus on gastric ulcer,two acute gastric ulcer mice models were established by intragastric administration of aspirin or absolute ethanol,respectively.Clinical index of ulcer area,ulcer index,gastric volume,gastric pH value,free acidity,total acidity,and histopathological assessment were measured to evaluate the injuries of gastric ulcer and the protective effect of okra seed oil In order to comprehensively uncover the possible underlying mechanism,a series of biochemical assays were also performed,including serum TNF-α,IL-6,IL-10 and Tbil,NO,MPO and SOD in the stomach included.Moreover,the ALT,AST and ALP in the liver of mice were also tested to evaluate the possible hepatic toxicity of the seed oil.The results indicated that the seed oil of A.esculentus exerted protective effect in ethanol-induced gastric ulcer mice by reducing the ulcer area and ulcer index,declining the free and total acidity,and increasing the pH value of gastric content.Histopathological observation showed the gastric mucosa of the acute gastric ulcer mice induced by alcohol was incomplete and severely damaged,with submucosal edema and nuclear pyknosis,as well as glandular structure disappearing,compared with that of normal mice.What's more,a number of inflammatory cell infiltration occured in the gastric mucosa of alcohol-model mice,with messes of neutrophils,lymphocytes,eosinophils and plasma cells.Okra seed oil could improve the damaged structure of the gastric mucosa and gland caused by ethanol,but could not ameliorate the condensation of nucleus and infiltration of inflammatory cells.Biochemical analysis revealed that the seed oil of A.esculentus could counteract the damage induced by ethanol via decreasing Tbil and TNF-α in serum,decreasing NO and myeloperoxidase,and increasing SOD in stomach.Meanwhile,okra seed oil exhibited protective effect in aspirin-induced gastric ulcer mice by increasing the gastric content pH,and reducing free and total acidity.Compared with the control group,the gastric mucosa of aspirin-model group showed multifocal coagulation necrosis,sheet edema and infiltration of inflammatory cells by histopathological assessment.Compared with the aspirin-model group,the soybean oil group and okra seed oil group could ameliorate the inflammatory cell infiltration.Biochemical analysis revealed that okra seed oil could counteract the injury induced by aspirin via decreasing TNF-α and IL-6,and increasing IL-1O in serum,decreasing NO and MPO and increasing SOD in stomach.In a word,the okra seed oil exerted protective effect on acute gastric ulcer by anti-inflammation,anti-oxidation and hepatocyte protection.The okra seed oil deserves further development and utilization.

Objective To investigate the effect of FCGR3A polymorphisms on NK cell function. Methods Peripheral blood samples from can?cer patients were collected and FCGR3A polymorphisms were confirmed by PCR. In vitro proliferation rates,ADCC activity,and expression of NK cell activating receptors were compared under trastumab stimulation. Results This study showed that the wild?type FCGR3A exhibited a higher affinity to trastumab along with better NK cell proliferation and ADCC activity than the mutant type. Compared to the patients with wild?type FC?GR3A,the proliferation rates of NK cells in patients with the mutant type were reduced by approximately 8?fold. In addition,the expression of NK cell activating receptors in patients with wild?type FCGR3A was higher than in patients with the mutant type. Conclusion Mutations in FC?GR3Areduce NK cell function,causing a poor reaction to monoclonal antibody.