1.Effects of Modified shaoyao gancao decoction on intestinal transit function,intestinal flora and metabolite content in slow transit constipation rat
Ziqi ZHANG ; Hongyun ZHOU ; Qiong ZHAO ; Yuan DENG ; Yu ZHAN
China Pharmacy 2025;36(2):154-159
OBJECTIVE To observe the effects of Modified shaoyao gancao decoction on intestinal transit function, intestinal flora and the contents of metabolites [γ aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT)] in slow transit constipation (STC) rats. METHODS SD rats were randomly divided into blank group (10 rats) and modeling group (30 rats), with half male and half female. The STC model was established by intragastric administration of Compound diphenoxylate tablets in the modeling group. The successfully modeled rats were randomly divided into model group, Modified shaoyao gancao decoction group [56 g/(kg·d), calculated by crude drug] and positive control group [lactulose 2.09 g/(kg·d)], with 10 rats in each group. Each administration group was given relevant medicine intragastrically, the blank group and model group received an equivalent volume of normal saline, once a day, for 14 consecutive days. During the experiment, the general situation of rats was observed in each group. After the last medication, the body weight was measured, and the Bristol score was used to evaluate the fecal characteristics. The fecal moisture content, intestinal propulsion rate, and the contents of GABA and 5-HT in intestinal content were detected; the diversity of intestinal flora in intestinal contents was investigated, and the correlation between the contents of GABA, 5-HT and relative abundance of microbiota was analyzed. RESULTS Compared with the model group, general conditions such as small body shape, sparse and rough fur, and slow movement were all improved in Modified shaoyao gancao decoction body weight, Bristol score, fecal moisture content,intestinal propulsion rate, 5-HT content, Chao1 index and Shannon index were increased significantly, while GABA content and Simpson index were decreased significantly (P<0.05). The intestinal flora of rats in the Modified shaoyao gancao decoction group could be classified as the same as the blank group, but was far from the model group; the relative abundances of Desulfobacterota, Firmicutes and Bacteroidota in this group showed a tendency of pull back, but the differences were not statistically significant compared to model group (P>0.05). Desulfobacterota was an intergroup differential factor (P<0.05). The content of GABA was negatively correlated with the relative abundance of Bacteroidota, Cyanobacteria, Patescibacteria and Actinobacteriota (P<0.05). The content of 5-HT was positively correlated with the relative abundance of Campilobacterota (P<0.05). CONCLUSIONS Modified shaoyao gancao decoction can improve the fecal properties and intestinal motility of STC rats. Its mechanism may be related to improving intestinal flora and then affecting the contents of GABA and 5-HT in intestinal contents. In addition, the contents of GABA and 5-HT may be significantly correlated with the relative abundance of specific bacterial phyla such as Bacteroidota and Campilobacterota.
2.Effects of Modified shaoyao gancao decoction on intestinal transit function,intestinal flora and metabolite content in slow transit constipation rat
Ziqi ZHANG ; Hongyun ZHOU ; Qiong ZHAO ; Yuan DENG ; Yu ZHAN
China Pharmacy 2025;36(2):154-159
OBJECTIVE To observe the effects of Modified shaoyao gancao decoction on intestinal transit function, intestinal flora and the contents of metabolites [γ aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT)] in slow transit constipation (STC) rats. METHODS SD rats were randomly divided into blank group (10 rats) and modeling group (30 rats), with half male and half female. The STC model was established by intragastric administration of Compound diphenoxylate tablets in the modeling group. The successfully modeled rats were randomly divided into model group, Modified shaoyao gancao decoction group [56 g/(kg·d), calculated by crude drug] and positive control group [lactulose 2.09 g/(kg·d)], with 10 rats in each group. Each administration group was given relevant medicine intragastrically, the blank group and model group received an equivalent volume of normal saline, once a day, for 14 consecutive days. During the experiment, the general situation of rats was observed in each group. After the last medication, the body weight was measured, and the Bristol score was used to evaluate the fecal characteristics. The fecal moisture content, intestinal propulsion rate, and the contents of GABA and 5-HT in intestinal content were detected; the diversity of intestinal flora in intestinal contents was investigated, and the correlation between the contents of GABA, 5-HT and relative abundance of microbiota was analyzed. RESULTS Compared with the model group, general conditions such as small body shape, sparse and rough fur, and slow movement were all improved in Modified shaoyao gancao decoction body weight, Bristol score, fecal moisture content,intestinal propulsion rate, 5-HT content, Chao1 index and Shannon index were increased significantly, while GABA content and Simpson index were decreased significantly (P<0.05). The intestinal flora of rats in the Modified shaoyao gancao decoction group could be classified as the same as the blank group, but was far from the model group; the relative abundances of Desulfobacterota, Firmicutes and Bacteroidota in this group showed a tendency of pull back, but the differences were not statistically significant compared to model group (P>0.05). Desulfobacterota was an intergroup differential factor (P<0.05). The content of GABA was negatively correlated with the relative abundance of Bacteroidota, Cyanobacteria, Patescibacteria and Actinobacteriota (P<0.05). The content of 5-HT was positively correlated with the relative abundance of Campilobacterota (P<0.05). CONCLUSIONS Modified shaoyao gancao decoction can improve the fecal properties and intestinal motility of STC rats. Its mechanism may be related to improving intestinal flora and then affecting the contents of GABA and 5-HT in intestinal contents. In addition, the contents of GABA and 5-HT may be significantly correlated with the relative abundance of specific bacterial phyla such as Bacteroidota and Campilobacterota.
3.Analysis of Mechanism of Xingpi Capsules in Treatment of Functional Dyspepsia Based on Transcriptomics
Rongxin ZHU ; Mingyue HUANG ; Keyan WANG ; Xiangning LIU ; Yinglan LYU ; Gang WANG ; Fangfang RUI ; Qiong DENG ; Jianteng DONG ; Yong WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):164-172
ObjectiveTo investigate the ameliorative effect of Xingpi capsules on functional dyspepsia(FD) and the potential mechanism. MethodsSixty SPF-grade male SD neonatal rats(7 days old) were randomly divided into the normal group(n=12) and the modeling group(n=48), and the FD model was prepared by iodoacetamide gavage in the modeling group. After the model was successfully prepared, the rats in the modeling group were randomly divided into the model group, the low-dose and high-dose groups of Xingpi capsules(0.135, 0.54 g·kg-1) and the domperidone group(3 mg·kg-1), with 12 rats in each group. Rats in the normal and model groups were gavaged with distilled water, and rats in the rest of the groups were gavaged with the corresponding medicinal solution, once a day for 7 d. The general survival condition of the rats was observed, and the water intake and food intake of the rats were measured, the gastric emptying rate and the small intestinal propulsion rate were measured at the end of the treatment, the pathological damage of the rat duodenum was examined by hematoxylin-eosin(HE) staining, and the expressions of colonic tight junction protein(Occludin) and zonula occludens protein-1(ZO-1) were detected by immunofluorescence. The differentially expressed genes in the duodenal tissues of the model group and the normal group, and the high-dose group of Xingpi capsules and the model group were detected by transcriptome sequencing after the final administration, and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out. The transcriptomic results were validated by Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the active ingredients of Xingpi capsules were screened for molecular docking with the key targets. ResultsCompared with the normal group, the general survival condition of rats in the model group was poorer, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly reduced(P<0.05), inflammatory infiltration was seen in duodenal pathology, and the fluorescence intensities of Occludin and ZO-1 in the colon were significantly reduced(P<0.01). Compared with the model group, the general survival condition of rats in the high-dose group of Xingpi capsules improved significantly, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly increased(P<0.05), the duodenal pathology showed a decrease in inflammatory infiltration, and the fluorescence intensities of colonic Occludin and ZO-1 were significantly increased(P<0.01). Transcriptomic results showed that Xingpi capsules might exert therapeutic effects by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) through the key genes such as Slc5a1, Abhd6. The validation results showed that compared with the normal group, the phosphorylation levels of PI3K and Akt proteins, the protein expression level of interleukin(IL)-1β, and the fluorescence intensities of IL-6 and IL-1β were significantly increased in the model group(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3, Slc5a9 and other key genes were significantly increased(P<0.01). Compared with the model group, the phosphorylation levels of PI3K and Akt, the protein expression level of IL-1β and the fluorescence intensities of IL-6 and IL-1β in the high-dose group of Xingpi capsules were significantly reduced(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3 and Slc5a9 were significantly reduced(P<0.05). Weighted gene co-expression network analysis and molecular docking results showed that E-nerolidol and Z-nerolidol in Xingpi capsules were well bound to ABDH6 protein, and linarionoside A, valerosidatum and senkirkine were well bound to Slc5a1 protein. ConclusionXingpi capsules can effectively improve the general survival and gastrointestinal motility of FD rats, its specific mechanism may be related to the inhibition of PI3K/Akt signaling pathway to alleviate the low-grade inflammation of duodenum, and E-nerolidol, Z-nerolidol, linarionoside A, valerosidatum and senkirkine may be its key active ingredients.
4.Analysis of Mechanism of Xingpi Capsules in Treatment of Functional Dyspepsia Based on Transcriptomics
Rongxin ZHU ; Mingyue HUANG ; Keyan WANG ; Xiangning LIU ; Yinglan LYU ; Gang WANG ; Fangfang RUI ; Qiong DENG ; Jianteng DONG ; Yong WANG ; Chun LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):164-172
ObjectiveTo investigate the ameliorative effect of Xingpi capsules on functional dyspepsia(FD) and the potential mechanism. MethodsSixty SPF-grade male SD neonatal rats(7 days old) were randomly divided into the normal group(n=12) and the modeling group(n=48), and the FD model was prepared by iodoacetamide gavage in the modeling group. After the model was successfully prepared, the rats in the modeling group were randomly divided into the model group, the low-dose and high-dose groups of Xingpi capsules(0.135, 0.54 g·kg-1) and the domperidone group(3 mg·kg-1), with 12 rats in each group. Rats in the normal and model groups were gavaged with distilled water, and rats in the rest of the groups were gavaged with the corresponding medicinal solution, once a day for 7 d. The general survival condition of the rats was observed, and the water intake and food intake of the rats were measured, the gastric emptying rate and the small intestinal propulsion rate were measured at the end of the treatment, the pathological damage of the rat duodenum was examined by hematoxylin-eosin(HE) staining, and the expressions of colonic tight junction protein(Occludin) and zonula occludens protein-1(ZO-1) were detected by immunofluorescence. The differentially expressed genes in the duodenal tissues of the model group and the normal group, and the high-dose group of Xingpi capsules and the model group were detected by transcriptome sequencing after the final administration, and Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analyses were carried out. The transcriptomic results were validated by Western blot, immunofluorescence, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR), and the active ingredients of Xingpi capsules were screened for molecular docking with the key targets. ResultsCompared with the normal group, the general survival condition of rats in the model group was poorer, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly reduced(P<0.05), inflammatory infiltration was seen in duodenal pathology, and the fluorescence intensities of Occludin and ZO-1 in the colon were significantly reduced(P<0.01). Compared with the model group, the general survival condition of rats in the high-dose group of Xingpi capsules improved significantly, and the water intake, food intake, gastric emptying rate and small intestinal propulsion rate were all significantly increased(P<0.05), the duodenal pathology showed a decrease in inflammatory infiltration, and the fluorescence intensities of colonic Occludin and ZO-1 were significantly increased(P<0.01). Transcriptomic results showed that Xingpi capsules might exert therapeutic effects by regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) through the key genes such as Slc5a1, Abhd6. The validation results showed that compared with the normal group, the phosphorylation levels of PI3K and Akt proteins, the protein expression level of interleukin(IL)-1β, and the fluorescence intensities of IL-6 and IL-1β were significantly increased in the model group(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3, Slc5a9 and other key genes were significantly increased(P<0.01). Compared with the model group, the phosphorylation levels of PI3K and Akt, the protein expression level of IL-1β and the fluorescence intensities of IL-6 and IL-1β in the high-dose group of Xingpi capsules were significantly reduced(P<0.05, P<0.01), and the mRNA levels of Slc5a1, Abhd6, Mgam, Atp1a1, Slc7a8, Cdr2, Chrm3 and Slc5a9 were significantly reduced(P<0.05). Weighted gene co-expression network analysis and molecular docking results showed that E-nerolidol and Z-nerolidol in Xingpi capsules were well bound to ABDH6 protein, and linarionoside A, valerosidatum and senkirkine were well bound to Slc5a1 protein. ConclusionXingpi capsules can effectively improve the general survival and gastrointestinal motility of FD rats, its specific mechanism may be related to the inhibition of PI3K/Akt signaling pathway to alleviate the low-grade inflammation of duodenum, and E-nerolidol, Z-nerolidol, linarionoside A, valerosidatum and senkirkine may be its key active ingredients.
5.Effects of Combined Tongue Three-Needle Acupuncture and Acupoint Application on Lianquan (CV 23) on Swallowing Function and Surface Electromyography Signals in Patients with Dysphagia after Ischemic Stroke
Xiaoyu DENG ; Dongmei XU ; Qiong FAN ; Lei YUAN ; Wei WU ; Haimei LIU
Journal of Traditional Chinese Medicine 2025;66(15):1559-1565
ObjectiveTo observe the clinical effectiveness and potential mechanism of combined tongue three-needle acupuncture and acupoint application on Lianquan (CV 23) for patients with dysphagia after ischemic stroke. MethodsA prospective study was conducted on 160 patients with post-stroke dysphagia, who were randomly divided into a treatment group and a control group, with 80 cases in each group. The control group received conventional rehabilitation training, while the treatment group received tongue three-needle acupuncture combined with acupoint application on Lianquan (CV 23) on the basis of conventional rehabilitation training, for 4 weeks in both groups. We compared the clinical effectivenss of both groups after treatment, and assessed the swallowing function including videofluoroscopic swallowing study (VFSS), standardized swallowing assessment (SSA) and functional oral intake scale (FIOS), swallowing contrast test including hyoid maximum displacement (HmaxD), pharyngeal transit time (PTT), and upper esophageal sphincter (UES) opening, surface electromyography (sEMG) test including maximum amplitude and swallowing duration as well as swallowing quality of life questionnaire (SWAL-QOL) score of the patients in both groups before treatment, after 2 weeks and 4 weeks of treatment, respectively. ResultsThe total effective rate in treatment group was 82.50% (66/80), significantly higher than 66.25% (53/80) in control group (P<0.05). The VFSS, and FOIS scores, UES opening rate and HmaxD, sEMG maximal amplitude values, and SWAL-QOL scores were increased in both groups after 2 weeks and 4 weeks of treatment compared with the values before treatment (P<0.05), while SSA scores, PTT, and swallowing duration were decreased compared within group before treatment (P<0.05). VFSS and FOIS scores, UES opening rate and HmaxD, sEMG maximal amplitude values, and SWAL-QOL scores after 2 and 4 weeks of treatment in the treatment group were higher (P<0.05), while SSA scores, PTT, and swallowing duration were lower (P<0.05) than those in the control group at the same time. ConclusionCombined tongue three-needle acupuncture and acupoint application on Lianquan (CV 23) for patients with dysphagia after ischemic stroke can significantly improve swallowing activities, and its mechanism of action may be related to the improvement of the contraction ability and coordination of swallowing-related muscle groups.
6.Study on the mechanism of Shaoyao gancao decoction in improving intestinal motility in rats with slow transit constipation by regulating the ASIC3/ERK signaling pathway
Ziqi ZHANG ; Hongyun ZHOU ; Qiong ZHAO ; Yuan DENG ; Mengjie ZHAO ; Chen ZHAO ; Jingyi CHEN
China Pharmacy 2025;36(15):1852-1858
OBJECTIVE To explore the mechanism of Shaoyao gancao decoction in improving intestinal motility in rats with slow transit constipation (STC) by regulating acid-sensitive ion channel 3 (ASIC3)/extracellular signal-regulated kinase (ERK) signaling pathway. METHODS SD rats were used to construct an STC model by gavage with compound diphenoxylate. The successfully modeled rats were randomly divided into model group, Shaoyao gancao decoction group (1.5 g/mL), lactulose group (208.4 mg/mL, positive control), and combined inhibition group (Shaoyao gancao decoction 1.5 g/mL+amiloride hydrochloride 20 μg/kg), with 12 rats in each group. Additionally, 12 healthy rats were selected as the blank group. They were given relevant medicine once a day and continuously intervened for 14 days. After intervention, the intestinal propulsion function and visceral sensitivity of the model rats were detected. The expression of ASIC3 in the colon tissue of rats was observed by immunohistochemical staining. mRNA expressions of ASIC3, ERK1 and ERK2 as well as protein expressions of ASIC3, ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in colon tissue of rats were detected; the ultrastructural changes of the enteric nervous system (ENS) -interstitial cell of Cajal (ICC)-smooth muscle cell (SMC) network in the rat colon were observed under electron microscopy. RESULTS Compared with the model group, the intestinal propulsion rate of the Shaoyao gancao decoction group was significantly increased, while the visceral pain threshold was significantly decreased. The proportion of the positive area of ASIC3 in the colonic tissue was significantly increased. The relative mRNA expression levels of ERK1, ERK2, and ASIC3, as well as the relative protein expression levels of p-ERK1/2 and ASIC3, and the p-ERK1/2 to ERK1/2 in the colonic tissue, were all significantly increased (P<0.05 or P<0.01). Additionally, there was marked repair of the morphological structure of ICC and SMC, with closer gap junctions observed. Compared with the Shaoyao gancao decoction group, the combined inhibition group exhibited a diminished improvement in intestinal motility of rats, with statistically significant differences in the levels of some indicators (P<0.05 or P<0.01); the repairing of the morphological structure of ICC and SMC was notably attenuated. CONCLUSIONS Shaoyao gancao decoction can effectively improve the intestinal transmission function and promote intestinal repair in STC rats, and its mechanism may be related to regulating the balance of the ENS-ICC-SMC network mediated by the ASIC3/ERK signaling pathway, thus promoting intestinal motility and reducing visceral sensitivity.
7.Oral Absorption and Labeling Techniques of Traditional Chinese Medicine Polysaccharides: A Review
Weifeng ZHU ; Shuangyan DENG ; Hui OUYANG ; Wenjing YANG ; Jianing FU ; Huangqing WEI ; Qiong LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(12):261-269
Polysaccharides are the important material basis of traditional Chinese medicine(TCM), and have various pharmacological activities such as immunomodulation, antitumor and anti-aging. Due to the large molecular weight of TCM polysaccharides, their structural analysis and oral absorption mechanism are facing technical challenges, and the current research on their structure-activity relationships has made some breakthroughs, while the research on their oral absorption mechanisms is relatively slow. In-depth study of the oral absorption mechanism of TCM polysaccharides is not only crucial for the interpretation of their action pathways and efficacy in vivo, but also helpful for the interpretation of their pharmacological effects, rational clinical applications and the discovery of new targets. In recent years, the application of fluorescent labeling and isotopic labeling methods has provided new technical means for the oral absorption studies of polysaccharides, which has promoted the development of oral absorption studies of TCM polysaccharides. In this paper, we reviewed the oral absorption pathways and labeling techniques of TCM polysaccharides, and concluded that they can be absorbed orally through transmembrane, cellular bypass, and M-cell-mediated transport, of which transmembrane pathway is the main absorption pathway, and summarized the labeling reactions of four fluorescent labeling and isotopic labeling methods with TCM polysaccharides, which can provide references for evaluating the absorption pathways of TCM polysaccharides, screening active TCM polysaccharides, establishing pharmacodynamic models and comprehensively elucidating the mechanism of TCM polysaccharides.
8.Clinical Analysis of Sangxing Zhike Prescription for the Treatment of Postinfectious Cough of Warm Dryness Invading the Lung Type
Zi-Xing LIU ; Yin-He DENG ; Meng-Zhao LI ; Yu-Jing CHEN ; Qiong LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(4):856-861
Objective To observe the clinical efficacy and safety of Sangxing Zhike Prescription in treating postinfectious cough(PIC)of warm dryness invading the lung type.Methods A total of 66 PIC patients with warm dryness invading the lung type who were admitted to the First Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2020 to June 2022 were randomly divided into a treatment group and a control group,with 33 patients in each group.The treatment group was given Sangxing Zhike Prescription combined with Compound Methoxyphenamine Capsules,and the control group was given Compound Methoxyphenamine Capsules combined with Chinese medicine placebo.The course of treatment covered 7 days.The changes in the Visual Analogue Scale(VAS)scores of the severity of cough,the scores of cough symptom,and the scores of traditional Chinese medicine(TCM)syndrome in the two groups were observed before and after the treatment.After treatment,the clinical efficacy and safety in the two groups were evaluated.Results(1)During the trial,one case fell off from the treatment group and 4 cases fell off from the control group,and eventually 61 cases completed the observation,of which 32 cases were in the treatment group and 29 cases were in the control group.(2)After 7 days of treatment,the total effective rate of the treatment group was 84.38%(27/32)and that of the control group was 58.62%(17/29),and the intergroup comparison(tested by chi-square test)showed that the therapeutic efficacy of the treatment group was significantly superior to that of the control group(P<0.05).(3)After treatment,the VAS scores of the severity of cough,and the scores of daytime cough,nighttime cough of the Cough Symptom Score Scale as well as the overall cough scores in the two groups were significantly lower than those before treatment(P<0.05 or P<0.01),and the reduction of the VAS scores and the overall cough symptom scores in the treatment group was significantly superior to that in the control group(P<0.05).(4)After treatment,obvious improvement was presented in the scores of TCM symptoms of cough,throat itching,dry throat,foreign body sensation in the throat,sore throat and pharyngeal signs as well as total TCM syndrome scores in the treatment group when compared with the pre-treatment period(P<0.01),whereas in the control group,only the scores of cough,throat itching,dry throat,and sore throat and the total TCM syndrome scores were improved compared with the pre-treatment period(P<0.05 or P<0.01).The post-treatment intergroup comparison showed that the treatment group was significantly superior to the control group in improving the scores of throat itching,foreign body sensation in the throat,and pharyngeal signs as well as total TCM syndrome scores(P<0.05 or P<0.01).(5)During the treatment process,no significant adverse reactions occurred in both groups,or no abnormal changes were shown in the safety indexes such as blood routine test,liver and kidney functions of the patients.Conclusion Sangxing Zhike Prescription combined with Compound Methoxyphenamine Capsules exerts certain effect in treating patients with PIC of warm dryness invading the lung type,and its efficacy is significantly superior to that of Compound Methoxyphenamine Capsules treatment alone with relatively high safety profile.
9.Expression of nuclear receptor NURR1 in prostate cancer and its effects on the circRNA profile
Hongliang WANG ; Hui LIANG ; Qiong DENG ; Ying ZHANG ; Zhu WANG
Journal of Modern Urology 2024;29(3):261-267
【Objective】 To investigate the expression profile of circRNA in nuclear receptor NURR1 overexpressed prostate cancer (PCa) cells, so as to provide reference for revealing the mechanism of PCa progression. 【Methods】 The expression of NURR1 in PCa was analyzed with UALCAN and TNMplot. The distinct circRNAs in NURR1 overexpressed PCa cells were screened with RNA-sequencing. The functions and signal pathways of differentially expressed circRNA molecules were analyzed with GO and KEGG. 【Results】 The circ_0000915 was significantly downregulated in DU145, LNCaP and PC3 cells. In NURR1 overexpressed DU145 cells, circ_0005991 was up-regulated, while circ_0001460 and circ_0001315 were down-regulated. In NURR1 overexpressed LNCaP cells, circ_0040729 and circ_0000722 were significantly up-regulated. In NURR1 overexpressed PC3 cells, circ_0001577, circ_0000854 and circ_0018168 were up-regulated, while circ_013035, circ_0003028, circ_0082096 and circ_0005320 were down-regulated. KEGG analysis revealed that the differentially expressed circRNAs were significantly associated with dorsal/ventral neural tube patterns, protein folding chaperones, disordered domain specific binding, positive regulation of BMP signaling pathways, and neural tube patterning functions. 【Conclusion】 CircRNAs play an important role in NURR1 mediated PCa progression, but there are certain differences among different prostate cancer cell types. The regulatory mechanism between NURR1 and circ_0000915 in the progression of PCa needs further investigation.
10.Establishment of Q-PCR method for qualitative testing of the presence or absence of KIR genes
Yunan LI ; Jianxin ZHEN ; Shuang LIANG ; Qiong YU ; Zhihui DENG
Chinese Journal of Blood Transfusion 2024;37(6):660-665
Objective To establish a method for qualitative detection of the presence or absence of all KIR genes by quantitative polymerase chain reaction(Q-PCR).Methods Based on the polymorphism of high-resolution level KIR alleles in Chinese population and the IPD-KIR database,KIR gene-specific primers were designed to amplify all the 16 KIR genes and 2DS4-Normal and 2DS4-Deleted subtypes by Q-PCR.Meanwhile,one negative control and one positive control specific amplifying human growth hormone(HGH)gene fragment were set to monitor the false positive and false negative results in PCR amplification,respectively.A total of 302 samples with known KIR genotype previously identified by KIR PCR-SSP commercial kit were randomly selected for blind inspection to verify the reliability of KIR Q-PCR method established by au-thors.Results The results of 300 samples detected by our KIR Q-PCR method were consistent with the known results,but two samples showed inconsistent results.One sample was negative for 2DS5 by Q-PCR but positive by PCR-SSP,another sample was positive for 2DS1 by Q-PCR but negative by PCR-SSP.The two doubtful samples were genotyped by sequencing-based typing(PCR-SBT)for 2DS5and2DS1,respectively.PCR-SBT results confirmed that the results of Q-PCR test was correct.Conclusion The KIR Q-PCR method established in this paper can provide accurate and reliable results for testing the presence or absence of KIR genes.

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