ObjectiveTo investigate the mechanism of Zuogui Wan （ZGW） in improving ovarian inflammatory microenvironment and stemness of ovarian germline stem cells （OSCs） for treating ovarian aging via the cyclic guanosine monophosphate/adenosine monophosphate synthase （cGAS）/stimulator of interferon genes （STING） signaling pathway. MethodsForty C57BL/6 female mice were randomly divided into a blank group， a model group， a low-dose ZGW group （2.7 g·kg^-1）， a high-dose ZGW group （5.4 g·kg^-1）， and an estradiol valerate group （0.15 mg·kg^-1）， with 8 mice in each group. Except the blank group， all other groups received a single intraperitoneal injection of cyclophosphamide at 120 mg·kg^-1 to establish an ovarian aging mouse model. After successful modeling， each group was continuously administered for 4 weeks， once daily. The physiological status of the mice was observed， and the ovarian index was calculated. The estrus cycle of the mice was monitored. Hematoxylin-eosin （HE） staining was used to observe pathological changes in ovarian tissue. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum sex hormone levels. Serum inflammatory factors interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and mouse interleukin-6 （IL-6） levels were detected using kits. Western blot was used to detect the protein expression of ovarian cGAS， STING， p-STING， TANK-binding kinase 1 （TBK1）， p-TBK1， interferon-induced transmembrane protein 3 （Fragilis）， and Vasa homolog protein （MVH）. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of inflammatory factors in ovarian tissue. Immunofluorescence double labeling was performed to locate OSCs in ovarian tissues， and fluorescence intensities of OSCs markers MVH and octamer binding transcription factor 4 （Oct4） were calculated. ResultsCompared with the blank group， the model group showed reduced body weight， ovarian wet weight， and ovarian index （P<0.01） and a disordered estrus cycle （P<0.01）. In addition， the levels of serum follicle-stimulating hormone （FSH）， TNF-α， IL-6， and IL-1β were increased （P<0.01）， while anti-Müllerian hormone （AMH） and estradiol （E₂） levels were decreased （P<0.01）. The protein expression of cGAS， p-STING/STING， and p-TBK1/TBK1 in ovarian tissue was increased （P<0.05， P<0.01）， while that of OSCs stemness factors MVH and Fragilis was reduced （P<0.01）. Immunofluorescence indicated a reduction in MVH and Oct4 expression in OSCs （P<0.01）. The mRNA expression of inflammatory factors TNF-α， IL-6， and IL-1β in ovarian tissue was increased （P<0.05， P<0.01）. Compared with the model group， the treatment groups exhibited improved body weight， ovarian wet weight， and ovarian index （P<0.05） and a reduced rate of estrus cycle disorder （P<0.05， P<0.01）. The levels of serum FSH， TNF-α， IL-6， and IL-1β were decreased （P<0.05， P<0.01）， while AMH and E₂ levels were increased （P<0.01）. The protein expression levels of cGAS， p-STING/STING， and p-TBK1/TBK1 in ovarian tissue were decreased （P<0.05）， while the protein expression of MVH and Fragilis was increased （P<0.05）， and the fluorescence intensities of MVH and Oct4 were increased （P<0.05， P<0.01）. The mRNA expression of inflammatory factors in ovarian tissue was decreased （P<0.05）. ConclusionZGW alleviate ovarian inflammatory response， regulate ovarian microenvironment homeostasis， and maintain stemness of OSCs in ovarian aging mice probably by modulating the cGAS-STING signaling pathway， thereby improving ovarian function and delaying ovarian aging.

OBJECTIVE:To develop a method for simultaneous determination of 6 alkaloids in Coptis teeta.METHODS:The determination was performed on XtimateTM C18 with mobile phase consisted of 30 mmol/L ammonium bicarbonate [containing 0.1％ triethylamine and 0.7％ ammonia-acetonitrile (gradient elution)] at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,the column temperature was 30 ℃,and the sample size was 10 μL.RESULTS:The linear ranges of jateorrhizine,columbamine,epiberberine,coptisine,palmatine hydrochloride and berberine hydrochloride were 0.85-16.96 mg/L (r=0.999 9),1.25-24.96 mg/L(r=0.999 8),2.05-40.96 mg/L(r=0.999 9),3.65-72.96 mg/L(r=0.999 9),2.88-57.60 mg/L(r=0.999 9) and 13.25-264.96 mg/L(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.Recoveries were 97.14％-102.14％ (RSD=1.93％,n=6),97.00％-102.00％ (RSD=2.06％,n=6),98.18％-101.82 ％ (RSD=1.79％,n=6),96.15％-101.28％ (RSD=2.06％,n=6),96.88％-101.88％ (RSD=1.87％,n=6),99.31％-103.76％ (RSD=1.89％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stability and reproducible,and can be used for simultaneous determination of 6 alkaloids in Coptis teeta.

Objective To develop a novel colorimetric method for detecting the tumor biomarker vascular endothelial growth factor (VEGF) based on aptamer and magnetic beads.Methods The capture aptamer was hybridized to urease functionalized single-stranded DNA (ssDNA) and immobilize on the surface of magnetic beads by specific biotin-avidin binding.In the presence of VEGF,aptamers bound to VEGF to form a specific stem-loop structure to release the urease functionalized ssDNA.After separation,the supernatant was transferred to a tube and urea and phenol red were added.Urease hydrolyzed urea to produce ammonia to cause an increase of the pH value and a color change of phenol red.The results were inspected with either the naked eyes or by a UV spectrophotometer.Results Under optimized conditions,the detection system showed a good linear relationship for VEGF detection in the range of 0.1 to 10 pmol/L with a detection limit as low as 0.06 pmol/L.The results of VEGF detection in the serum of patients with lung cancer were consistent with those using an ELISA Kit.The results of examination of 10 serum samples with this aptamer-based method and ELISA kit showed that the accuracy of this method was 90％.Conclusion This aptamer-based system provides an simple and convenient method for VEGF detection with a high sensitivity and selectivity.

Objective To develop a novel colorimetric method for detecting the tumor biomarker vascular endothelial growth factor (VEGF) based on aptamer and magnetic beads.Methods The capture aptamer was hybridized to urease functionalized single-stranded DNA (ssDNA) and immobilize on the surface of magnetic beads by specific biotin-avidin binding.In the presence of VEGF,aptamers bound to VEGF to form a specific stem-loop structure to release the urease functionalized ssDNA.After separation,the supernatant was transferred to a tube and urea and phenol red were added.Urease hydrolyzed urea to produce ammonia to cause an increase of the pH value and a color change of phenol red.The results were inspected with either the naked eyes or by a UV spectrophotometer.Results Under optimized conditions,the detection system showed a good linear relationship for VEGF detection in the range of 0.1 to 10 pmol/L with a detection limit as low as 0.06 pmol/L.The results of VEGF detection in the serum of patients with lung cancer were consistent with those using an ELISA Kit.The results of examination of 10 serum samples with this aptamer-based method and ELISA kit showed that the accuracy of this method was 90％.Conclusion This aptamer-based system provides an simple and convenient method for VEGF detection with a high sensitivity and selectivity.

AIM : To set up a method of determinting the contents of four iridoid glycosides in different preparations of Lamiophlomis rotata ( Benth.) kudo.METHODS : The quantitative analysis was carded out on a column of Waters Symmetry by HPLC using a mobile phase of CH_3OH-H_2O under a flow rate of 1.0 mL/min.235 nm was selected as the detective wavelength.RESULTS: The linear ranges were 0.236-1.18 μg for sesamoside,0.440-2.20 μg for shanzhiside methyl ester,0.094-0.470 μg for 7,8-dehydropenste moside、0.750-3.75 μg for 8-O-ace-tylshanzhiside methyl ester.The recoveries of them were all between 96% and 104% ( RSD ＜ 5% ).CONCLUSION: The method is accurate,stable,reliable,and can be used as a quantitative standard for determining the contents of iridoid glycosides in different Duyiwei preparations.

Aim To explore the mechanisms of the apoptosis induction and the effects of adhesion suppression of Zhiling capsule (ZLJN) in small cell lung cancer cell line NCI-H446.Methods According to the different components of ZLJN,NCI-H446 cells were treated with traditional Chinese medicine,western medicine and ZLJN composite groups.Apoptotic cells were tested by light microscopy,Hochest33258 staining method.The mRNA and protein expressions of bcl-2,bax and hTERT were analyzed by RT-PCR and Western blot respectively.The expressions of CD44 were detected by flow cytometry.Results After NCI-H446 cells were treated with different drug groups,The morphological changes of apoptotic cells were found by light microscopy and Hochest33258 staining method.The mRNA and protein expressions of bcl-2 were down-regulated while the expressions of bax were up-regulated compared to the control groups(P

Humans ; Severe Acute Respiratory Syndrome ; diagnosis ; therapy

Aim To investigate the effects of Zhiling capsule (ZLJN) on the proliferation inhibition and apoptosis induction in K562 cell line.Methods According to the different components of ZLJN,K562 cells were treated respectively with tradtional Chinese medicine,Western medicine and ZLJN compound groups.The cell viability and colony formation were observed by MTT assay and colony formation assay respectively.Apoptotic cells were detected by Annexin V-FITC/PI staining and DNA fragmentation assay.Caspase-3 activity was detected by flow cytometry,and pro-caspase-3 was detected by Western blot.Results Treated with drug,K562 cell growth and cell colony formation were significantly inhibited.Apoptosis occurring in the early stage was identified by Annexin V-FITC/PI staining.Typical DNA ladder was seen from gel electrophoresis and apparent apoptotic peaks were observed by flow cytometer.The level of caspase-3 activity increased after the treatment,while the level of pro-caspase-3 decreased.Conclusion ZLJN can efficiently inhibit proliferation and induce apoptosis in K562 cells,which may be related with the up-regulation of caspase-3 activity.

AIM:To set up a method of determinting the contents of four iridoid glycosides in different preparations of Lamiophlomis rotata(Benth.)kudo.METHODS:The quantitative analysis was carried out on a column of Waters Symmetry by HPLC using a mobile phase of CH_3OH-H_2O under a flow rate of 1.0 mL/min.235 nm was selected as the detective wavelength.RESULTS:The linear ranges were 0.236-1.18 ?g for sesamoside,0.440-2.20 ?g for shanzhiside methyl ester、0.094-0.470 ?g for 7,8-dehydropenste moside、0.750-3.75 ?g for 8-O-acetylshanzhiside methyl ester.The recoveries of them were all between 96% and 104%(RSD