ObjectiveTo explore the molecular mechanism by which gypenoside L （Gyp-L） promotes apoptosis and inhibits ovarian cancer （OC） through the FK506-binding protein （FKBP） prolyl isomerase 8 （FKBP8）/B-cell lymphoma-2 （Bcl-2） axis， with the piR-hsa-2804461 pathway as a breakthrough point. MethodsThe effects of different concentrations of Gyp-L and cis-platinum on the proliferation of OVCAR3 cells were determined by the cell count kit-8 method to identify the appropriate intervention concentration for subsequent experiments. OVCAR3 cells were allocated into blank， low-dose Gyp-L （Gyp-L-L， 50 µmol·L^-1）， high-dose Gyp-L （Gyp-L-H， 100 µmol·L^-1）， and cis-platinum （15 µmol·L^-1） groups. The migration， colony formation， and apoptosis of OVCAR3 cells were detected by the cell scratch assay， colony formation assay， and flow cytometry， respectively. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes in OVCAR3 cells were determined by Real-time PCR， and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by simple Western blot. Further， an OVCAR3 cell model with piR-hsa-2804461 knocked out was constructed. The cells were allocated into blank， NC-inhibitor， inhibitor， NC-inhibitor+Gyp-L， and inhibitor+Gyp-L groups. The colony formation of OVCAR3 cells was detected by the colony formation assay. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by Real-time PCR and simple Western blotting， respectively. ResultsGyp-L inhibited the migration and proliferation （P<0.01）， promoted the apoptosis （P<0.05）， up-regulated the mRNA level of piR-hsa-2804461 （P<0.05）， and down-regulated the mRNA and protein levels of FKBP8 and Bcl-2 （P<0.05） in OVCAR3 cells. Furthermore， Gyp-L increased the mRNA and protein levels of Bcl-2-associated X protein （Bax）， cysteinyl aspartate-specific proteinase （Caspase）-3， and Caspase-9， which are related to the FKBP8/Bcl-2 axis （P<0.05）. ConclusionGyp-L may promote apoptosis by regulating the piR-hsa-2804461/FKBP8/Bcl-2 axis， thus affecting the occurrence of ovarian cancer.

ObjectiveTo explore the molecular mechanism of gypenoside L （Gyp-L） in the treatment of ovarian cancer （OC） by taking the glycolytic pathway of OC as the key point. MethodsThe proliferation activity of OVCAR3 cells was measured by the cell counting kit-8 （CCK-8） assay to determine the appropriate intervention concentration for subsequent experiments. The cell clone formation assay and the scratch healing assay were employed to assess the proliferation and migration capabilities of OVCAR3 cells. OVCAR3 cells were divided into a blank group， a Gyp-L-L group （low concentration of Gyp-L， 50 µmol·L^-1）， a Gyp-L-H group （high concentration of Gyp-L， 100 µmol·L^-1）， and a cisplatin （15 µmol·L^-1） group. The glucose consumption in OC cells was determined using a kit， and the expression levels of related mRNAs in OVCAR3 cells were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expressions of related proteins were detected by Wes automatic Western blot quantitative analysis. ResultsValidated by the cell scratch assay， the scratch healing rate of OVCAR3 cells was decreased after Gyp-L intervention， reflecting that Gyp-L could significantly inhibit the migration of OVCAR3 cells （P<0.05）. According to the clone formation assay， the cell colony formation ability of the Gyp-L-L group， Gyp-L-H group， and cisplatin group was significantly lower than that of the control group （P<0.05）. In OVCAR3 cells， compared with those in the control group， the levels of glucose consumption and lactate generation in the Gyp-L-L group and Gyp-L-H group were significantly reduced （P<0.05）， indicating that Gyp-L could effectively inhibit the glycolysis level of OC cells. Meanwhile， Gyp-L could suppress the expression levels of glucokinase （GCK）， phosphofructokinase liver （PFKL）， signal transducer and activator of transcription 3 （STAT3）， lactate dehydrogenase D （LDHD）， phosphoglycerate mutase 1 （PGAM1）， mRNAs， and proteins related to the glycolytic pathway in OC cells. ConclusionGyp-L may inhibit the occurrence and development of OC by influencing the GCK-mediated glycolytic pathway.

ObjectiveWith the epidermal growth factor receptor（EGFR）/Signal Transducers and Activators of Transcription（STAT3）/Hexokinase 2（HK2） signaling pathway in atherosclerosis （AS） and ovarian cancer （OC） as the entry point， this paper discusses the molecular mechanism of Gypenoside L （Gyp-L） treating AS and OC with different diseases， provides a new perspective and theoretical basis for TCM treating AS and OC with EGFR-STAT3-HK2 pathway， and enriches the scientific connotation of the theory of "cytoskeleton in the heart". MethodsCCK-8 was used to detect the proliferation of HUVEC and OVCAR-3 cells， in order to determine the intervention concentration for subsequent experiments. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. Western blot was used to detect the expression levels of relevant proteins. Furthermore， two cell models overexpressing EGFR were constructed and co treated with Gyp-L. HUVEC cells were divided into control， ox-LDL， OE-NC， OE-EGFR， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. OVCAR-3 cells were divided into control， OE-NC， OE-EGFR ， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Western blot was used to detect the expression levels of EGFR-STAT3-HK2 pathway related proteins. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. ResultsGyp-L can significantly reduce the NO content of HUVEC and the pyruvate and LDH content of two cell lines （P<0.05）； Inhibit the proliferation and migration ability of OVCAR-3 cells； Reduce the expression levels of EGFR/STAT3/HK2 pathway related proteins in HUVEC and OVCAR-3 cell lines （P<0.05）， and inhibit the glycolysis pathway. ConclusionGyp-L can inhibit glycolysis in HUVEC and OVCAR-3 cells through the EGFR/STAT3/HK2 pathway，thereby suppressing the occurrence and development of AS and OC.

ObjectiveTo explore the molecular mechanism by which gypenoside L （Gyp-L） promotes apoptosis and inhibits ovarian cancer （OC） through the FK506-binding protein （FKBP） prolyl isomerase 8 （FKBP8）/B-cell lymphoma-2 （Bcl-2） axis， with the piR-hsa-2804461 pathway as a breakthrough point. MethodsThe effects of different concentrations of Gyp-L and cis-platinum on the proliferation of OVCAR3 cells were determined by the cell count kit-8 method to identify the appropriate intervention concentration for subsequent experiments. OVCAR3 cells were allocated into blank， low-dose Gyp-L （Gyp-L-L， 50 µmol·L^-1）， high-dose Gyp-L （Gyp-L-H， 100 µmol·L^-1）， and cis-platinum （15 µmol·L^-1） groups. The migration， colony formation， and apoptosis of OVCAR3 cells were detected by the cell scratch assay， colony formation assay， and flow cytometry， respectively. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes in OVCAR3 cells were determined by Real-time PCR， and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by simple Western blot. Further， an OVCAR3 cell model with piR-hsa-2804461 knocked out was constructed. The cells were allocated into blank， NC-inhibitor， inhibitor， NC-inhibitor+Gyp-L， and inhibitor+Gyp-L groups. The colony formation of OVCAR3 cells was detected by the colony formation assay. The mRNA levels of piR-hsa-2804461 and FKBP8/Bcl-2 axis-related genes and the expression levels of FKBP8/Bcl-2 axis-related proteins were determined by Real-time PCR and simple Western blotting， respectively. ResultsGyp-L inhibited the migration and proliferation （P<0.01）， promoted the apoptosis （P<0.05）， up-regulated the mRNA level of piR-hsa-2804461 （P<0.05）， and down-regulated the mRNA and protein levels of FKBP8 and Bcl-2 （P<0.05） in OVCAR3 cells. Furthermore， Gyp-L increased the mRNA and protein levels of Bcl-2-associated X protein （Bax）， cysteinyl aspartate-specific proteinase （Caspase）-3， and Caspase-9， which are related to the FKBP8/Bcl-2 axis （P<0.05）. ConclusionGyp-L may promote apoptosis by regulating the piR-hsa-2804461/FKBP8/Bcl-2 axis， thus affecting the occurrence of ovarian cancer.

ObjectiveTo explore the molecular mechanism of gypenoside L （Gyp-L） in the treatment of ovarian cancer （OC） by taking the glycolytic pathway of OC as the key point. MethodsThe proliferation activity of OVCAR3 cells was measured by the cell counting kit-8 （CCK-8） assay to determine the appropriate intervention concentration for subsequent experiments. The cell clone formation assay and the scratch healing assay were employed to assess the proliferation and migration capabilities of OVCAR3 cells. OVCAR3 cells were divided into a blank group， a Gyp-L-L group （low concentration of Gyp-L， 50 µmol·L^-1）， a Gyp-L-H group （high concentration of Gyp-L， 100 µmol·L^-1）， and a cisplatin （15 µmol·L^-1） group. The glucose consumption in OC cells was determined using a kit， and the expression levels of related mRNAs in OVCAR3 cells were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expressions of related proteins were detected by Wes automatic Western blot quantitative analysis. ResultsValidated by the cell scratch assay， the scratch healing rate of OVCAR3 cells was decreased after Gyp-L intervention， reflecting that Gyp-L could significantly inhibit the migration of OVCAR3 cells （P<0.05）. According to the clone formation assay， the cell colony formation ability of the Gyp-L-L group， Gyp-L-H group， and cisplatin group was significantly lower than that of the control group （P<0.05）. In OVCAR3 cells， compared with those in the control group， the levels of glucose consumption and lactate generation in the Gyp-L-L group and Gyp-L-H group were significantly reduced （P<0.05）， indicating that Gyp-L could effectively inhibit the glycolysis level of OC cells. Meanwhile， Gyp-L could suppress the expression levels of glucokinase （GCK）， phosphofructokinase liver （PFKL）， signal transducer and activator of transcription 3 （STAT3）， lactate dehydrogenase D （LDHD）， phosphoglycerate mutase 1 （PGAM1）， mRNAs， and proteins related to the glycolytic pathway in OC cells. ConclusionGyp-L may inhibit the occurrence and development of OC by influencing the GCK-mediated glycolytic pathway.

ObjectiveWith the epidermal growth factor receptor（EGFR）/Signal Transducers and Activators of Transcription（STAT3）/Hexokinase 2（HK2） signaling pathway in atherosclerosis （AS） and ovarian cancer （OC） as the entry point， this paper discusses the molecular mechanism of Gypenoside L （Gyp-L） treating AS and OC with different diseases， provides a new perspective and theoretical basis for TCM treating AS and OC with EGFR-STAT3-HK2 pathway， and enriches the scientific connotation of the theory of "cytoskeleton in the heart". MethodsCCK-8 was used to detect the proliferation of HUVEC and OVCAR-3 cells， in order to determine the intervention concentration for subsequent experiments. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. Western blot was used to detect the expression levels of relevant proteins. Furthermore， two cell models overexpressing EGFR were constructed and co treated with Gyp-L. HUVEC cells were divided into control， ox-LDL， OE-NC， OE-EGFR， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. OVCAR-3 cells were divided into control， OE-NC， OE-EGFR ， OE-NC+Gyp-L， and OE-EGFR+Gyp-L group. The colorimetric method was used to detect the NO content in HUVEC and the contents of pyruvate and LDH in two cell lines. Western blot was used to detect the expression levels of EGFR-STAT3-HK2 pathway related proteins. Cell cloning experiments and scratch experiments reflect the proliferation and migration ability of OVCAR-3 cells. ResultsGyp-L can significantly reduce the NO content of HUVEC and the pyruvate and LDH content of two cell lines （P<0.05）； Inhibit the proliferation and migration ability of OVCAR-3 cells； Reduce the expression levels of EGFR/STAT3/HK2 pathway related proteins in HUVEC and OVCAR-3 cell lines （P<0.05）， and inhibit the glycolysis pathway. ConclusionGyp-L can inhibit glycolysis in HUVEC and OVCAR-3 cells through the EGFR/STAT3/HK2 pathway，thereby suppressing the occurrence and development of AS and OC.

AIM: To quantify early changes of macular capillary parameters in type 2 diabetic patients using optical coherence tomography angiography(OCTA).METHODS: Retrospective case study. A total of 49 healthy subjects, 52 diabetic patients without retinopathy(noDR)patients, and 43 mild nonproliferative diabetic retinopathy(mNPDR)patients were recruited. Capillary perfusion density, vessel length density(VLD), and average vessel diameter(AVD)were calculated from macular OCTA images(3 mm×3 mm)of the superficial capillary plexus after segmenting large vessels and the deep capillary plexus. Parameters were compared among control subjects, noDR, and mNPDR patients. The area under the receiver operating characteristic curve estimated the abilities of these parameters to detect early changes of retinal microvascular networks.RESULTS: Significant differences were found in the VLD and AVD among the three groups(P<0.001). Compared with the control group, the noDR group had significantly higher AVD(P<0.05). VLD of both layers in patients of mNPDR group was significant decreased compared with that of noDR group(all P<0.01). Deep AVD had a higher area under the curve(AUC)of 0.796 than other parameters to discriminate the noDR group from the healthy group. Deep AVD had the highest AUC of 0.920, followed by that of the deep VLD(AUC=0.899)to discriminate the mNPDR group from the healthy group.CONCLUSIONS: NoDR patients had wider AVD than healthy individuals and longer VLD than mNPDR patients in both layers. When compared with healthy individuals, deep AVD had a stronger ability than other parameters to detect early retinal capillary impairments in noDR patients.

Objective To observe the clinical effect and safety of recombinant human interferon-α2b(rhIFN-α2b)combined with 5-aminolaevulinic acid photodynamic therapy(ALA-PDT)in the treatment of patients with cervical intraepithelial lesions and human papilloma virus(HPV)infection.Methods The clinical data of patients with cervical intraepithelial lesions and HPV infection were analyzed retrospectively.The patients were divided into control group and treatment group according to cohort method.The control group was treated with rhIFN-α2b gel at posterior fornix,qd.On this basis,the treatment group was treated with ALA-PDT,namely applying 5％ALA temperature-sensitive gel prepared by 118 mg of ketone valerate hydrochloride powder for external use on vaginal surface,combined with photodynamic therapy,once a week.The clinical efficacy,time to recovery from clinical symptoms,recurrence rate,changes in cytokines[interleukin-6(IL-6),interleukin-8(IL-8)and interferon-γ(IFN-γ)]and immune function[T cell subsets CD3+,CD4+,CD8+and CD4+/CD8+],and safety were compared between the two groups.Results There were 98 cases in treatment group and 100 cases in control group.The total effective rates in treatment group and control group were 93.88％and 83.00％,with statistically significant difference(P＜0.05).The relief time of lower abdominal pain in treatment group and control group were(3.65±0.52)and(5.26±0.65)d;the time to recovery from abnormal vaginal discharge were(5.77±0.83)and(7.16±0.92)d;the time to recovery from irregular vaginal bleeding were(4.82±0.62)and(6.94±0.77)d;HPV clearance rates were 58.16％and 42.00％;IL-6 levels were(0.16±0.09)and(0.23±0.05)mg·L-1;IL-8 levels were(0.47±0.05)and(0.66±0.07)mg·L-1;IFN-γ levels were(10.07±0.98)and(7.24±0.65)ng·mL-1;CD3+were(71.06±8.29)％and(61.36±6.88)％;CD4+were(48.25±5.94)％and(42.25±5.13)％;CD8+were(20.37±2.42)％and(24.69±2.51)％;CD4+/CD8+were 2.11±0.27 and 1.36±0.16;the differences were all statistically significant(all P＜0.05).Recurrence rates in treatment group and control group were 5.10％and 14.00％,with statistically significant difference(P＜0.05).The total incidence of adverse drug reactions in treatment group and control group were 26.53％and 21.00％,without statistically significant difference(P＞0.05).Conclusion The combined treatment of rhIFN-α2b and ALA-PDT is effective for patients with cervical intraepithelial lesions and HPV infection.It has obvious advantages in improving immunity,relieving inflammatory response and reducing recurrence rate,with good safety.

The traditional commodity specifications of Chinese medicinal materials are mainly divided into different grades based on macroscopic characteristics. As the basis for high quality and good price, there is still a lack of systematic evaluation on whether they are consistent with the current standards and whether they can reflect the internal quality of medicinal material. Panax notoginseng is a commonly used, large consumption of Chinese medicinal material. At present, it is divided into 8 grades in the market based on "Tou" (the number of crude drug /500 g), but it is not related to the standard of total saponins of Panax notoginseng (the sum of three saponins) in Chinese Pharmacopoeia. In this study, ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) coupled with mass spectrometry molecular network were used for the rapid identification of saponins of Panax notoginseng with different "Tou" and a total of 64 saponins were identified. Seventeen saponins related to "Tou" were screened by orthogonal partial least squares discriminant analysis (OPLS-DA). The content of five saponins R₁, Rb₁, Rg₁, Rd, and Re in Panax notoginseng with different "Tou" was determined by high performance liquid chromatography (HPLC). The results of correlation analysis showed that Rd and R₁ with the largest VIP values among the differential saponins, which significantly negatively correlated with "Tou" (P < 0.05). Based on the determination results of 36 batches of samples, using Rd/Total Panax notoginseng saponins (TPNS) ratio ( > 0.08) as the index, Panax notoginseng can be divided into two grades: 20-60 "Tou" (superior) and 80-200 "Tou" (qualified). Based on the concept of "macroscopic characteristics and chemical profiling", this study integrates the non-targeted analysis and quantitative determination methods to provide a new strategy for quality evaluation of Panax notoginseng.

[摘 要] 目的：探讨人参皂苷Rb1（Gn-Rb1）对肝细胞癌（HCC）HepG2细胞氧死亡的影响及其可能的分子机制。方法：采用生物信息学方法分析氧死亡的关键基因PGAM5表达对HCC患者生存期的影响。选取辽宁省肿瘤医院收治的8例HCC患者的HCC组织与癌旁组织，通过WB法及qPCR检测氧死亡相关基因蛋白与mRNA的表达情况。将HepG2细胞随机分为对照组与Gn-Rb1组（予以200 μmol/L Gn-Rb1干预），采用细胞克隆形成实验、划痕愈合实验分别检测Gn-Rb1对HepG2细胞的集落形成能力、迁移能力的影响，ELISA检测对细胞ROS生成水平的影响，微板法检测对细胞LDH释放水平的影响；WB法、qPCR法检测Gn-Rb1对HepG2氧死亡关键基因蛋白质与mRNA水平表达的影响。结果：生物信息学分析发现，PGAM5高表达肝癌患者总生存时间较低表达患者更长（P<0.05）。在临床HCC组织与癌旁组织样本中发现，相较于癌旁组织，在蛋白质与mRNA水平上，肿瘤组织KEAP1与PGAM5表达显著降低，NRF2表达显著升高（均P<0.01），p-AIFM1蛋白水平显著升高（P<0.05）。对HepG2细胞予以200 μmol/L Gn-Rb1干预后，相较于对照组，Gn-Rb1组HepG2细胞的迁移能力与集落形成能力显著降低（均P<0.01），而LDH水平显著升高（P<0.05）；相比于对照组，在mRNA和蛋白质水平上，Gn-Rb1组细胞中KEAP1、PGAM5表达均显著升高而NRF2表达均显著降低（均P<0.05），p-AIFM1蛋白表达显著降低（P<0.01）。结论：HCC组织中氧死亡被抑制，而Gn-Rb1能够通过调控KEAP1/PGAM5/AIFM1通路促进HepG2细胞氧死亡的发生，抑制细胞增殖和迁移能力。