1.Study on Quality Standard of Chailing Hugan Granules
Guodong MO ; Jin YANG ; Li LIN ; Fangting TAN
China Pharmacy 2018;29(20):2796-2800
OBJECTIVE:To establish the quality standard of Chailing hugan granules. METHODS:TLC was used for qualitative identification of Radix bupleuri,Atractylodes macrocephala and Glycyrrhiza uralensis. The content of total flavonoids (by rutin) in preparation was determined by UV-visible spectrophotometry. The moisture, dissolubility and granularity of preparation were determined. RESULTS:TLC spots of R. bupleuri ,A. macrocephala and G. uralensis were clear and well-separated without interference from negative control. The linear range of rutin was 0.050-0.300 mg/mL(r=0.999 8). RSDs of precision, stability and reproducibility tests were all lower than 2%. The recoveries were 97.89%-100.01%(RSD=0.68%,n=9). The content of samples were 1.920-2.018 mg/g. The contents of moisture in 3 batches of samples were 1.54%,1.62% and 1.57%;all samples dissolved within 5 min. The sum of granules not passing through No.1 sieve and passing through No.5 sieve were 2.13%, 2.51%,2.38%,which were all in line with the requirements of Chinese Pharmacopeia (2015 edition,Vol Ⅳ). CONCLUSIONS:The content of total flavonoicle in Chailing hugan granules should be no less than 1.57 mg/g (by rutin). Established quality standard is simple,rapid,accurate and reproducible,and can be used for quality control of Chailing hugan granules.
2.The effect of salpingectomy with preservation of the mesosalpinx on ovarian reserve function
Bo LI ; Chuancui HU ; Da ZHAO ; Fangting LIN ; Yan GAO ; Caiyun LIN ; Xiaoli WANG
Journal of Clinical Medicine in Practice 2024;28(4):34-38
Objective To investigate the impact of salpingectomy with preservation of fallopian tube mesentery on ovarian reserve function. Methods A total of 281 patients with tubal disease who were admitted from January 2020 to March 2021 were collected as research objects. According to the different treatment methods, they were divided into traditional bilateral resection group (
3.Karyotype analysis and fluorescence in situ hybridization detection of 122 patients with myelodysplastic syndrome.
Yuan JING ; Shuang LIN ; Feng JIANG ; Fangting WANG ; Meiyun FANG
Chinese Journal of Medical Genetics 2016;33(2):221-226
OBJECTIVETo assess the value of karyotype analysis and fluorescence in situ hybridization(FISH) assay for the diagnosis of myelodysplastic syndrome (MDS).
METHODSThe karyotypes of 122 initially treated MDS patients were analyzed with conventional R-banding and FISH using probes including GLP CSF1R/D5S23, D5S721, GLP EGR1/D5S23, D5S721, GLP D7S486/CSP7, GLP D7S522/CSP7, GLP D20S108, CSP8 and CSP X/Y.
RESULTSThe detection rate of chromosomal abnormalities was 54.9% for the 122 patients. Among these, those involving 3 or more chromosomes are most common (16.4%), followed by +8(14.8%), -7/7q-(7.4%), -5/5q-(5.7%), 20q-(2.5%), and -Y in male patients (5.0%). Two MDS-RAEB II patients detected with t(8;21) should be diagnosed with acute myelocytic leukemia. FISH analysis showed that 54 patients were positive (44.3%). Among these, 30.3% had CSP8 amplification, followed by GLP D7S486/CSP7 and GLP D7S522/CSP7 deletion (12.3%), GLP CSF1R/D5S23, D5S721 and GLP EGR1/D5S23, D5S721 deletion (9.8%), GLP D20S108 deletion (7.4%), and CSPX/Y deletion (5%).
CONCLUSIONWith a detection rate of 54.9%, R-banding still constitutes the basic examination for MDS. As detection of interstitial chromosomal abnormalities in MDS can be greatly enhanced by FISH, combined karyotype analysis and FISH can improve the diagnosis of MDS and facilitate assessment of its prognosis.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromosome Aberrations ; Chromosome Banding ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Sequence Deletion ; Young Adult
4.Inhibition of NLRP3 inflammasome activation improves PCOS granulosa cell apoptosis by regulating autophagy
Shanhua FU ; Lili BAO ; Da ZHAO ; Jun LI ; Fangting LIN ; Rong HU
Chinese Journal of Immunology 2024;40(8):1646-1652
Objective:To investigate the expression of NOD-like receptor protein 3(NLRP3)inflammasome and its relation-ship with granulosa cell apoptosis in patients with polycystic ovary syndrome(PCOS).Methods:Follicular fluid and ovarian granulosa cells were collected from 17 PCOS patients(PCOS group)and 20 non-PCOS patients(control group),and the expression levels of pro-inflammatory factor,TNF-α,IL-1β and IL-18 in follicular fluid were detected by ELISA.RT-PCR and Western blot were used to de-tect NLRP3 mRNA and NLRP3 inflammasome associated protein NLRP3,ASC and cleaved caspase-1 and autophagy related protein LC3-Ⅱ/LC3-Ⅰ and p62 expression level in granulosa cells.TUNEL assay was used to detect the apoptotic level of granulosa cells in both groups.NLRP3 silenced siRNA(si-NLRP3)and negative control sequence(si-NC)were transfected into human ovarian cancer granulosa cell line KGN cells by siRNA interference technique in vitro.TNF-α was used to simulate PCOS-related cell damage.KGN cells were divided into 4 groups according to different treatment methods:Ctrl group,TNF-α group,TNF-α+si-NLRP3 group and TNF-α+si-NC group.The levels of DHEA,testosterone,IL-1β and IL-18 in supernatant were detected by ELISA.TUNEL assay was used to detect the apoptosis level of KGN cells.The LC3-Ⅱ/LC3-Ⅰ and p62,NLRP3,ASC and cleaved caspase-1 protein expression levels,and NF-κB p-p65 level(NF-κB p-p65/NF-κB p65)in KGN cells were detected by Western blot.Results:Compared with con-trol group,the concentration of TNF-α,IL-1β,and IL-18 in follicular fluid of PCOS patients,and LC3-Ⅱ/LC3-Ⅰ,NLRP3 mRNA and the protein expression of NLRP3,ASC,and cleaved caspase-1 in granulosa cells of and apoptosis were significantly increased(P<0.05 or P<0.01),while the protein expression of p62 was significantly decreased(P<0.01).Compared with Ctrl group,the levels of DHEA,testosterone,IL-1β and IL-18 in supernatant of TNF-α group,TNF-α+si-NC group,and TNF-α+si-NLRP3 group,and LC3-Ⅱ/LC3-Ⅰ,ASC and cleaved caspase-1 protein expression,and the level of NF-κB p-p65 and apoptosis were significantly increased(P<0.05 or P<0.01),while p62 protein was significantly decreased(P<0.01).NLRP3 was significantly decreased except in TNF-α+si-NLRP3 group(P<0.01),while in the TNF-α group and TNF-α+si-NC group was significantly increased(P<0.01).However,com-pared with TNF-α group,the above indexes in TNF-α+si-NLRP3 group were significantly decreased(P<0.05),and TNF-α+si-NC group had no significant change(P>0.05).Conclusion:The over-activated NLRP3 inflammasome in granulosa cells may promote cell inflammatory injury and autophagy apoptosis in PCOS patients through NF-κB pathway.
6.Analgesic Effect and Mechanism of Osteoking on Dorsal Root Ganglion Compression Model Rats
Wuqiong HOU ; Ying ZONG ; Fangting ZHOU ; Sensen LI ; Liting XU ; Xueying TAO ; Chunfang LIU ; Chao WANG ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):30-41
ObjectiveTo investigate the analgesic effect and mechanism of Osteoking (OK) on nerve compression in lumbar disc herniation. MethodThe rat model of chronic compression of dorsal root ganglion (CCD) was established to simulate clinical lumbar disc herniation. The CCD rats were randomly divided into model group, low, medium, and high dose OK groups (1.31, 2.63, 5.25 mL·kg-1·d-1), and pregabalin group (5 mg·kg-1), with eight rats in each group. Another eight SD rats were taken as the blank group, and the same volume of normal saline was given by gavage. Behavioral tests, side effect evaluation, network analysis, Western blot, immunofluorescence, and antagonist application were used to explore the effect. ResultCompared with the blank group, the mechanical hyperalgesia threshold, thermal hyperalgesia threshold, and the expression of inflammatory factors in the spinal dorsal horn of the model group are significantly increased (P<0.01), and the related indicators of the affected foot footprints are significantly down-regulated (P<0.01). The expression of signal transducer and activator of transcription 3 (STAT3), vascular endothelial growth factor A (VEGFA), and phosphorylated extracellular regulated protein kinase (p-ERK) in microglia in the spinal dorsal horn is significantly increased in the model group (P<0.01). Compared with the model group, low, medium, and high dose OK groups can increase the mechanical hyperalgesia and thermal hyperalgesia thresholds of CCD rats (P<0.05, P<0.01) in a dose-dependent manner, improve the gait of CCD rats (P<0.05, P<0.01), and reduce the expression of inflammatory factors in the spinal dorsal horn (P<0.05, P<0.01). The expression of STAT3, VEGFA, and p-ERK in the spinal dorsal horn microglia of CCD rats is significantly decreased (P<0.05, P<0.01), and the acetic acid-induced nociceptive response in rats is effectively reduced (P<0.05, P<0.01). In addition, there is no tolerance. The results of the body mass test, organ index, forced swimming, and rotation show that OK has no obvious toxic or side effects. Further antagonist experiments show that MRS1523 and RS127445 can reverse the transient analgesic effect of OK compared with the high dose OK group (P<0.01). ConclusionOK has a good analgesic effect on the CCD model without obvious toxic side effects, and its mechanism may be related to the activation of ADORA3 and HTR2B and the inhibition of STAT3, VEGFA, p-ERK, and other elements in microglia.