OBJECTIVE: Rhodiolae Crenulatae Radix et Rhizoma (Hongjingtian in Chinese, RCRR), the roots and rhizomes of Rhodiola crenulata and its application in the medicinal market is very chaotic. In this study, DNA barcoding database and identification engine of Rhodiola species were established, decoction pieces from the medicinal market were identified, and the application and challenges of DNA barcoding in the rapid radiation of Rhodiola species were analyzed. This study provides reference for the protection, rational development, and utilization of endangered resources within Rhodiola species. METHODS: A total of 50 original plant samples from 20 species of the genus Rhodiola from Hebei, Xinjiang, Tibet, Jilin, and other major production areas were collected. Theses samples cover the typical distribution area (Qinghai-Tibetan Platea) of Rhodiola species and other scattered alpine regions (Changbai Mountain, Taibai Mountain, Lushan Mountain, etc.), it encompasses all Rhodiola species with thick rhizomes in China. ITS2 and psbA-trnH barcode of Rhodiola database (BORD) were established and an identification engine named Rhodiola-IDE was developed. The stability and accuracy of the standard DNA barcoding database were evaluated using two datasets. Rhodiola-IDE identified 31 decoction pieces of RCRR from the medicinal material market. RESULTS: The BORD containing 1 532 sequences of 88 Rhodiola species has been established, and the identification efficiency results showed good accuracy and stability. According to the Chinese Pharmacopoeia (2020 edition), 23 samples (74.2%) were identified as authentic R. crenulata, while the rest of the marketed varieties were R. kirilowii, R. dumulosa, and R. fastigiata. The product label "Larger flower, Hongjingtian" was identified as R. crenulata. Samples labeled as "Smaller flower, Hongjingtian" were identified as R. crenulata, R. kirilowii, and R. fastigiata. CONCLUSION ITS2 and psbA-trnH barcodes can identify monophyletic groups represented by R. crenulata. However, for non-monophyletic species, it is necessary to collect as many samples as possible and combine them with multiple markers for joint identification. This study discussed the application and challenges of DNA barcodes in Rhodiola under rapid radiation conditions, providing a scientific basis for the rational development and utilization of Rhodiola varieties.

Abstract: Objective To investigate the function and molecular mechanism of LRH-1 in regulating the sensitivity of hepatocellular carcinoma (HCC) cells to oxaliplatin, providing new ideas for the treatment of liver cancer. Methods Knockdown and overexpression of LRH-1 in HCC cell lines were constructed, and the effect of LRH-1 on oxaliplatin resistance of HCC cells was explored by detecting IC50, cell proliferation, and plate colony formation assay. The transcriptional regulation of the MDR-1 gene by LRH-1 was detected through quantitative PCR. The transcriptional activation ability of LRH-1 on the MDR1 gene was evaluated by luciferase reporter assay. Results In HuH7 cells overexpressing LRH-1, the IC50 significantly increased to 18.012 μmol/L under oxaliplatin treatment, significantly higher than the 2.042 μmol/L in the HuH-7 control group, showing statistically significant differences (P<0.05). After overexpression of LRH-1 in HuH-7 cells, the cell proliferation ability was significantly increased, with a noticeable increase in MDR1 mRNA level. In HepG2 cells with knockdown LRH-1 expression, the IC50 significantly dropped to 1.012 μmol/L, significantly lower than the 6.294 μmol/L in the HepG2 control group, with statistically significant differences (P<0.05). After knockdown of LRH-1 in HepG2 cells, the cell proliferation and plate colony formation ability were significantly inhibited, with a notable decrease in MDR1 mRNA expression level. Luciferase reporter assay confirmed that LRH-1 can activate the transcriptional activity of the MDR1 promoter in a dosedependent manner, and its specific inhibitor ML-180 can significantly reduce LRH-1′s transcription activation ability on the MDR1 promoter. Conclusions LRH-1 may promote oxaliplatin resistance in hepatocellular carcinoma cells by regulating the transcriptional activity of MDR1 gene. Since its specific small molecule inhibitor has been successfully synthesized, LRH-1 can potentially become a target for the treatment of drug resistance in hepatocellular carcinoma.

Objective:To compare the short-term efficacy between our self-designed intelligent robot-assisted minimally invasive reduction system and conventional freehand reduction assisted by fluoroscopy in the treatment of unstable pelvic fractures by robot or fluoroscopy-assisted internal fixation with percutaneous screws.Methods:A prospective randomized controlled trial was conducted to include eligible 35 patients with unstable pelvic fracture who were admitted to Department of Orthopaedic Trauma, Beijing Jishuitan Hospital from December 2021 to October 2022. They were randomized into 2 groups. The observation group[17 cases, 10 males and 7 females with an age of (44.0±17.4) years] was treated with robot-assisted minimally invasive reduction, followed by robot-assisted or fluoroscopic internal fixation with percutaneous screws; the control group[18 cases, 12 males and 6 females with an age of (38.8±15.0) years] was treated with freehand reduction assisted by fluoroscopy, followed by robot-assisted or fluoroscopic internal fixation with percutaneous screws. The 2 groups were compared in terms of operation time, intraoperative bleeding, successful reduction, reduction quality, incidence of surgical complications and postoperative functional scores.Results:The 2 groups were comparable because there were no significant differences in the preoperative general data between them ( P>0.05). The intraoperative fluoroscopy frequency[(32.4±17.5) times] and fluoroscopy time [(19.8±10.4) s] in the observation group were significantly lower or shorter than those in the control group [(60.8±26.6) times and (38.2±16.1) s], and the rate of successful reduction in the observation group was 100.0% (17/17), significantly higher than that in the control group[72.2% (13/18)] ( P<0.05). There was no significant difference between the 2 groups in intraoperative bleeding, operation time, reduction error, excellent and good rate of reduction after operation by Matta scoring, or Majeed functional score at 12 weeks after operation ( P>0.05). Conclusion:In the treatment of unstable pelvic fractures, since our self-designed intelligent robot-assisted minimally invasive reduction system can plan autonomously the reduction paths and accomplish minimally invasive reduction of the fracture with 3D images real-time monitoring, it is advantageous over conventional reduction methods in a higher success rate and less radiation exposure.

Objective:To evaluate a self-designed intelligent robot-assisted minimally invasive reduction system in the reduction of unstable pelvic fractures by a cadaveric anatomic study.Methods:Ten unembalmed cadavers (7 male and 3 female ones) were used in this study. In each cadaveric specimen an unstable pelvic fracture was created in accordance with clinical case models (3 cases of type B1, 4 cases of type B2 and 3 cases of type C1 by the Tile classification). A self-designed intelligent robot-assisted minimally invasive reduction system was used to assist the reduction in the cadaveric models. Intraoperative registration and navigation time, autonomous reduction time, total operation time and reduction error were measured.Results:Effective reduction was completed in 10 bone models with the assistance of our self-designed intelligent robot-assisted minimally invasive reduction system. The time for intraoperative registration and navigation averaged 47.4 min (from 32 to 74 min), the autonomous reduction time 73.9 min (from 48 to 96 min), and the total operation time 121.3 min (from 83 to 170 min). The reduction error averaged 2.02 mm (from 1.67 to 2.62 mm), and the reduction results met the clinical requirements.Conclusion:Our self-designed intelligent robot-assisted minimally invasive reduction system is a new clinical solution for unstable pelvic fractures, showing advantages of agreement with clinical operative procedures, high reduction accuracy and operational feasibility, and reduced radiation exposure compared to a conventional operation.

PURPOSE: C-end rule (CendR) peptides are found to enhance the penetration of chemotherapeutic agents into tumor cells, while GX1 is a peptide that homes to gastric cancer (GC) vasculature. This study aimed to synthesize a novel peptide GX1-RPAKPAR (GXC) and to explore the effect of GXC on sensitizing GC cells to chemotherapeutic agents. MATERIALS AND METHODS: Intracellular Adriamycin concentration analysis was applied to conform whether GXC peptide increases the penetration of chemotherapeutic agents into GC cells in vitro. The effect of GXC peptide on sensitizing GC cells to chemotherapeutics was validated by apoptosis assay and in vitro/vivo drug sensitivity assay. The specificity of GXC to GC tissue was validated by ex vivo fluorescence imaging. RESULTS: In vitro, administration of GXC significantly increased Adriamycin concentrations inside SGC-7901 cells, and enhanced the efficacy of chemotherapeutic agents by decreasing the IC50 value. In vivo, FITC-GXC specifically accumulated in GC tissue. Moreover, systemic co-injection with GXC peptide and Adriamycin statistically improved the therapeutic efficacy in SGC-7901 xenograft models, surprisingly, without obviously increasing side effects. CONCLUSION: These results demonstrated that co-administration of the novel peptide GXC with chemotherapeutic agents may be a potential way to enhance the efficacy of anticancer drugs in GC treatment.
Apoptosis ; Doxorubicin ; Drug Therapy* ; Heterografts ; In Vitro Techniques ; Inhibitory Concentration 50 ; Optical Imaging ; Peptides ; Sensitivity and Specificity ; Stomach Neoplasms*

C.citratus has been usedin many countries with a long history.Traditionally,it is applied as a food seasoning in cooking.It is also used in tea beverage and folk medicine as well.Modern application of C.citratus is focused on the development of citronella oil,which can be used for food additives,disinfectants,cosmetics,drugs and etc.C.citratus is also a potential plant in landscaping.Its special lemony flavor contains chemical constituents,mainly including citral,myrcene,linalool,geraniol,nerol,citronellol,and etc.The modern research showed that C.citratus had the main effects of anti-microbial,anti-inflammation,analgesia,anti-oxidation,anti-tumor,anti-anxiety,anti-hypertension,antihyperglycemia,and etc.With further studies,some new pharmacological properties of C.citrates are going to be discovered gradually.It is worthy of further research and development to meet the needs of the health industry.

Objective: To study the chemical constituents in the dry roots of Dolomiaea souliei (Franch.) Shih.Methods: Various chromatographic methods were used to isolate and purify the chemical constituents of Dolomiaea souliei, and the structures were elucidated through the analysis of spectral data and literatures.Results: Six compounds including 3 sesquiterpene compounds and 3 fatty acids were obtained and identified as dihydrodehydrocostuslactone(Ⅰ), vladimenal(Ⅱ), arbusculin A(Ⅲ), n-hendecane(Ⅳ), butanedioic acid(Ⅴ) and methyl linoleate(Ⅵ).Conclusion: Compounds Ⅳ-Ⅵ are obtained from the genus of Dolomiaea for the first time.

Objective To discuss DNA methylation's effect on pathogenesis of pediatric immune thrombocytopenia (ITP)through detecting the expression level of DNA methyltransferases (DNMTs)mRNA in peripheral blood lymphocytes of children with ITP.Methods Two mL peripheral blood was collected from each of 25 children with persistent and chronic ITP and 20 healthy children (the healthy control group)by using aseptic method in the pediatric ward of the First Affiliated Hospital of Xinxiang Medical University from January 2014 to January 2015.First ethylene diamine tetraacetic acid (EDTA) was used as the anticoagulant.Then separate the mononuclear cells,extract RNA and detect expression levels of DNMT1,DNMT3A and DNMT3B mRNA using reverse transcription-polymerase chain reaction (RT-PCR) method.Results (1) The blood platelet (PLT) of children with persistent and chronic ITP was (36.2 ± 19.6) × 109/L,which was obviously lower than the healthy control group(168.8 ±46.8) × 109/L(t =-11.85,P =0.000).(2)The DNMT1 mRNA expression level of children with persistent and chronic ITP was 0.17 ± 0.05,which was obviously lower than the healthy control group (0.27 ± 0.10) (t =-3.912,P =0.001).The DNMT3A mRNA expression level of children with persistent and chronic ITP was 0.20 ± 0.10,which was obviously lower than the healthy control group (0.32 ±0.11) (t =-3.779,P =0.000).The DNMT3B mRNA expression level of children with persistent and chronic ITP was 0.16 ± 0.1 1,which was obviously lower than the healthy control group (0.31 ±0.11) (t =-4.641,P =0.000).(3) There was positive correlation between the expression of DNMT1 and DNMT3B mRNA(r =0.433,P =0.031).There was positive correlation between the expression of DNMT3A and DNMT3B mRNA(r =0.721,P =0.000).Conclusions (1) Children with persistent and chronic ITP have lower expression levels of DNMT1,DNMT3A,DNMT3 B mRNA,which indicates that DNA methylation contributes to the pathogenesis of pediatric persistent and chronic ITP.(2) DNMTs have synergistic effect on DNA methylation of pediatric persistent and chronic ITP.

Objective To analyze the functional changes and the clinical significance of B cell specific mono-clonal murine leukemia virus integration site -1 (Bmi -1 )and Th1 /Th2 cells in children with newly diagnosed im-mune thrombocytopenia(ITP)by testing the mRNA expressions of Bmi -1,helper T cell -related cytokine interferon (IFN)-γand interleukin(IL)-4 in children with newly diagnosed ITP.Methods Thirty -six cases of patients with newly diagnosed ITP in the experimental group came from the inpatient and outpatient children admitted to the Depart-ment of Pediatrics of the First Affiliated Hospital of Xinxiang Medical University from April to December 201 3.In the control group,26 cases of children requiring selective operation were admitted to the Department of Pediatric Surgery during the same period.The mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes were detected by means of the reverse transcription -polymerase chain reaction(RT -PCR)method,and were analyzed and compared by t test and linear correlation analysis.Results (1 )The mRNA expressions of Bmi -1,IFN -γand IL -4 in peripheral blood lymphocytes in the experimental group were 2.63 ±0.54,3.84 ±0.43 and 1 .44 ±0.39,respec-tively;while the mRNA expressions of Bmi -1,IFN -γand IL -4 in the peripheral blood lymphocytes in the control group were 3.91 ±0.92,2.88 ±0.57 and 1 .87 ±0.34,respectively.The levels of IFN -γof the experimental group were significantly higher than those of the control group (P <0.001 )and the levels of Bmi -1 and IL -4 in the experi-mental group were significantly lower than those in the control group (all P <0.001 ).(2)The mRNA expressions be-tween IFN -γand IL -4 in the peripheral blood lymphocytes in the experimental group were in negative correlation (r =-0.667,P <0.001 ).The mRNA expressions between IL -4 and Bmi -1 in the same group were in a positive correlation (r =0.776,P <0.001 ).There were no correlation in the mRNA expressions between IFN -γand Bmi -1 (r =-0.206,P >0.05).Conclusions Bmi -1 may be involved in the pathogenesis of ITP by regulating Th cell, and Th cell dysfunction may occur in the children with ITP,and the disproportion between Th1 and Th2 may be due to the advantages of Th1 .

OBJECTIVETo investigate the molecular mechanism of the growth inhibitory effect of matrine on K562 cells in JAK/STAT3 mediated signal pathway.

METHODSWestern blot analyses were performed to investigate the differential expression of JAK2, STAT3, phosphor-STAT3 (Tyr705 & Ser727) and phosphor-JAK2 proteins after matrine treatment in K562 cells with or without human recombinant interleukin 6 (IL-6) pretreatment. The expression of STAT3 response gene products such as Bcl-xL, Cyclin D1 and c-Myc, were investigated by Western blot and quantitative real time RT-PCR (qRT-PCR). Expression of IL-6, a potent upstream activating factor of JAK/STAT3 pathway, was analyzed by both real time qRT-PCR and ELISA.

RESUTLSWestern blot revealed that matrine treatment resulted in a strong down-regulation of phosphor-STAT3 both in Tyr705 and Ser727 sites or phosphor-JAK2 proteins expression without significant effects on the total STAT3 and JAK2 proteins. The expression of phosphor-Tyr705 STAT3 and phosphor-Ser727 STAT3 was decreased to 0.370 ± 0.172 in K562 cells treated with 0.5 mg/ml matrine for 48 h, respectively, from 0.690 ± 0.119 and 1.150 ± 0.263 in control cells, accompanied with a dramatical down-regulation of phosphor-JAK2 from 0.670 ± 0.137 to 0.049 ± 0.057 (P<0.05). In addition, it was found that the expression of Bcl-xL, Cyclin D1, c-Myc was decreased both at the transcription and protein level in K562 cells after matrine treatment. Matrine treatment resulted in a significant decrease in the expression level of IL-6 in K562 cells from (35.1 ± 1.93) to (10.74 ± 1.83) and (8.66 ± 1.24) pg/ml at the dose of 0.5 and 0.8 mg/ml, respectively (p<0.05). Matrine treatment could diminish the up-regulation of STAT3, JAK2, phosphor-STAT3 and phosphor-JAK2 protein following pretreatment with IL-6 in K562 cells.

CONCLUSIONMatrine exerts its anti-leukemia effect by interfering with the JAK2/STAT3 signaling pathway. The inhibition of IL-6 expression may play a pivotal role in the disruption of JAK/STAT pathway by matrine.

Alkaloids ; Down-Regulation ; Humans ; Interleukin-6 ; Janus Kinase 2 ; K562 Cells ; Quinolizines ; STAT3 Transcription Factor ; Signal Transduction ; Up-Regulation