Genetic variants, haplotype determination, and function of novel alleles of CYP2B6 in a Han Chinese population.

Amino acid variants in protein may result in deleterious effects on enzymatic activity. In this study we investigate the DNA variants on activity of CYP2B6 gene in a Chinese Han population for potential use in precision medicine. All exons in CYP2B6 gene from 1483 Chinese Han adults (Zhejiang province) were sequenced using Sanger sequencing. The effects of nonsynonymous variants on recombinant protein catalytic activity were investigated in vitro with Sf12 system. The haplotype of novel nonsynonymous variants with other single nucleotide variants in the same allele was determined using Nanopore sequencing. Of 38 alleles listed on the Pharmacogene Variation Consortium, we detected 7 previously reported alleles and 18 novel variants, of which 11 nonsynonymous variants showed lower catalytic activity (0.00-0.60) on bupropion compared to CYP2B6*1. Further, these 11 novel star-alleles (CYP2B6*39-49) were assigned by the Pharmacogene Variation Consortium, which may be valuable for pharmacogenetic research and personalized medicine.

[Clinical Efficacy and Safety of Ixazomib-Containing Regimens in the Treatment of Patients with Multiple Myeloma].

OBJECTIVE: To investigate the clinical efficacy and safety of ixazomib-containing regimens in the treatment of patients with multiple myeloma (MM). METHODS: A retrospective analysis was performed on the clinical efficacy and adverse reactions of 32 MM patients treated with a combined regimen containing ixazomib in the Hematology Department of the First People's Hospital of Lianyungang from January 2020 to February 2022. Among the 32 patients, 15 patients were relapsed and refractory multiple myeloma (R/RMM) (R/RMM group), 17 patients who responded to bortezomib induction therapy but converted to ixazomib-containing regimen due to adverse events (AE) or other reasons (conversion treatment group). The treatment included IPD regimen (ixazomib+pomalidomide+dexamethasone), IRD regimen (ixazomib+lenalidomide+dexamethasone), ICD regimen (ixazomib+cyclophosphamide+dexamethasone), ID regimen (ixazomib+dexamethasone). RESULTS: Of 15 R/RMM patients, overall response rate (ORR) was 53.3%(8/15), among them, 1 achieved complete response (CR), 2 achieved very good partial response (VGPR) and 5 achieved partial response (PR). The ORR of the IPD, IRD, ICD and ID regimen group were 100%（3/3）, 42.9%（3/7）, 33.3%（1/3）, 50%（1/2）， respectively， there was no statistically significant difference in ORR between four groups (χ 2=3.375, P =0.452). The ORR of patients was 50% after first-line therapy, 42.9% after second line therapy, 60% after third line therapy or more, with no statistically significant difference among them (χ2=2.164, P =0.730). In conversion treatment group, ORR was 88.2%(15/17), among them, 6 patients achieved CR, 5 patients achieved VGPR and 4 patients achieved PR. There was no statistically significant difference in ORR between the IPD（100%， 3/3）, IRD（100%， 6/6）, ICD（100%， 3/3） and ID（60%， 3/5） regimen groups (χ2=3.737，P =0.184). The median progression-free survival (PFS) time of R/RMM patients was 9 months (95% CI : 6.6-11.4 months), the median overall survival (OS) time was 18 months (95% CI : 11.8-24.4 months). The median PFS time of conversion treatment group was 15 months (95% CI : 7.3-22.7 months), the median OS time not reached. A total of 10 patients suffered grade 3- 4 adverse event (AE). The common hematological toxicities were leukocytopenia, anemia, thrombocytopenia. The common non-hematological toxicities were gastrointestinal symptoms (diarrhea, nausea and vomit), peripheral neuropathy, fatigue and infections. Grade 1-2 peripheral neurotoxicity occurred in 7 patients. CONCLUSION: The ixazomib-based chemotherapy regimens are safe and effective in R/RMM therapy, particularly for conversion patients who are effective for bortezomib therapy. The AE was manageable and safe.

Inhibitory effects of the main metabolites of Apatinib on CYP450 isozymes in human and rat liver microsomes.

PURPOSE: The inhibitory effect of Apatinib on cytochrome P450 (CYP450) enzymes has been studied. However, it is unknown whether the inhibition is related to the major metabolites, M1-1, M1-2 and M1-6. METHODS: A 5-in-1 cocktail system composed of CYP2B6/Cyp2b1, CYP2C9/Cyp2c11, CYP2E1/Cyp2e1, CYP2D6/Cyp2d1 and CYP3A/Cyp3a2 was used in this study. Firstly, the effects of APA and its main metabolites on the activities of HLMs, RLMs and recombinant isoforms were examined. The reaction mixture included HLMs, RLMs or recombinant isoforms (CYP3A4.1, CYP2D6.1, CYP2D6.10 or CYP2C9.1), analyte (APA, M1-1, M1-2 or M1-6), probe substrates. The reactions were pre-incubated for 5 min at 37 °C, followed by the addition of NAPDH to initiate the reactions, which continued for 40 min. Secondly, IC50 experiments were conducted to determine if the inhibitions were reversible. The reaction mixture of the "+ NADPH Group" included HLMs or RLMs, 0 to 100 of µM M1-1 or M1-2, probe substrates. The reactions were pre-incubated for 5 min at 37 °C, and then NAPDH was added to initiate reactions, which proceeded for 40 min. The reaction mixture of the "- NADPH Group" included HLMs or RLMs, probe substrates, NAPDH. The reactions were pre-incubated for 30 min at 37 °C, and then 0 to 100 µM of M1-1 or M1-2 was added to initiate the reactions, which proceeded for 40 min. Finally, the reversible inhibition of M1-1 and M1-2 on isozymes was determined. The reaction mixture included HLMs or RLMs, 0 to 10 µM of M1-1 or M1-2, probe substrates with concentrations ranging from 0.25Km to 2Km. RESULTS: Under the influence of M1-6, the activity of CYP2B6, 2C9, 2E1 and 3A4/5 was increased to 193.92%, 210.82%, 235.67% and 380.12% respectively; the activity of CYP2D6 was reduced to 92.61%. The inhibitory effects of M1-1 on CYP3A4/5 in HLMs and on Cyp2d1 in RLMs, as well as the effect of M1-2 on CYP3A in HLMs, were determined to be noncompetitive inhibition, with the Ki values equal to 1.340 µM, 1.151 µM and 1.829 µM, respectively. The inhibitory effect of M1-1 on CYP2B6 and CYP2D6 in HLMs, as well as the effect of M1-2 on CYP2C9 and CYP2D6 in HLMs, were determined to be competitive inhibition, with the Ki values equal to 12.280 µM, 2.046 µM, 0.560 µM and 4.377 µM, respectively. The inhibitory effects of M1-1 on CYP2C9 in HLMs and M1-2 on Cyp2d1 in RLMs were determined to be mixed-type, with the Ki values equal to 0.998 µM and 0.884 µM. The parameters could not be obtained due to the atypical kinetics of CYP2E1 in HLMs under the impact of M1-2. CONCLUSIONS: M1-1 and M1-2 exhibited inhibition for several CYP450 isozymes, especially CYP2B6, 2C9, 2D6 and 3A4/5. This observation may uncover potential drug-drug interactions and provide valuable insights for the clinical application of APA.

Puerarin Induces Ferroptosis in Colorectal Cancer Cells via Triggering NCOA4 Upregulation.

Puerarin shows promise as an anti-cancer compound, but its mechanism of action remains unclear. Here we explored whether and how it promotes ferroptosis in a colorectal cancer cell line. The level of ferroptosis and expression of autophagy proteins were compared between puerarin-treated HT-29 cells expressing normal or reduced levels of the autophagy protein ATG5 or the ferritinophagy protein nuclear receptor coactivator 4 (NCOA4). Puerarin increased lipid peroxidation and inhibited cell proliferation in a dose-dependent manner, indicating the induction of ferroptosis. These effects were partially reversed by ferrostatin-1, a scavenger of reactive oxygen species; by the iron chelator deferiprone; by repression of autophagy through administration of 3-methyladenine or knockdown of autophagy-related gene 5 (ATG5); or by repression of ferritinophagy through NCOA4 knockdown. Puerarin may induce the proliferative inhibition of colorectal cancer cells by triggering ferroptosis through a mechanism requiring NCOA4 ferritinophagy.

High expression of peroxisomal D-bifunctional protein in cytosol regulates apoptosis and energy metabolism of hepatocellular carcinoma cells via PI3K/AKT pathway.

Peroxisomal D-bifunctional protein (DBP) is an indispensable enzyme of the fatty acid ß-oxidation in the peroxisome of humans. However, the role of DBP in oncogenesis is poorly understood. Our previous studies have demonstrated that DBP overexpression promotes hepatocellular carcinoma (HCC) cell proliferation. In this study, we evaluated the expression of DBP in 75 primary HCC samples using RT-qPCR, immunohistochemistry, and Western blot, as well as its correlation with the prognosis of HCC. In addition, we explored the mechanisms by which DBP promotes HCC cell proliferation. We found that DBP expression was upregulated in HCC tumor tissues, and higher DBP expression was positively correlated with tumor size and TNM stage. Multinomial ordinal logistic regression analysis indicated that lower DBP mRNA level was an independent protective factor of HCC. Notably, DBP was overexpressed in the peroxisome and cytosol and mitochondria of tumor tissue cells. Xenograft tumor growth was promoted by overexpressing DBP outside peroxisome in vivo. Mechanistically, DBP overexpression in cytosol activated the PI3K/AKT signaling axis and promoted HCC cell proliferation by downregulating apoptosis via AKT/FOXO3a/Bim axis. In addition, overexpression of DBP increased glucose uptake and glycogen content via AKT/GSK3ß axis, as well as elevated the activity of mitochondrial respiratory chain complex III to increase ATP content via the mitochondrial translocation of p-GSK3ß in an AKT-dependent manner. Taken together, this study was the first to report the expression of DBP in peroxisome and cytosol, and that the cytosolic DBP has a critical role in the metabolic reprogramming and adaptation of HCC cells, which provides a valuable reference for instituting an HCC treatment plan.

[Efficacy and Safety of Decitabine Combined with Modified EIAG Regimen in the Treatment of Patients with Relapsed/Refractory Acute Myeloid Leukemia and High-risk Myelodysplastic Syndrome].

OBJECTIVE: To investigate the efficacy, prognosis and safety of decitabine combined with modified EIAG regimen in the treatment of patients with relapsed/refractory acute myeloid leukemia (AML) and high-risk myelodysplastic syndrome (MDS). METHODS: The clinical data of 44 patients with relapsed/refractory AML and high-risk MDS admitted to our hospital from January 2017 to December 2020 were analyzed retrospectively. The patients were equally divided into D-EIAG group (decitabine combined with EIAG regimen) and D-CAG group (decitabine combined with CAG regimen) according to clinical treatment regimen. The complete response (CR), CR with incomplete hematologic recover (CRi), morphologic leukemia-free state (MLFS), partial response (PR), overall response rate (ORR), modified composite complete response (mCRc), overall survival (OS) time, 1-year OS rate, myelosuppression and adverse reactions between the two groups were compared. RESULTS: In D-EIAG group, 16 patients (72.7%) achieved mCRc (CR+CRi+MLFS), 3 patients (13.6%) achieved PR, and ORR (mCRc+PR) was 86.4%. In D-CAG group, 9 patients (40.9%) achieved mCRc, 6 patients (27.3%) achieved PR, and ORR was 68.2%. Difference was observed in mCRc rate between the two groups (P=0.035), but not in ORR (P>0.05). The median OS time of D-EIAG group and D-CAG group was 20 (2-38) months and 16 (3-32) months, and 1-year OS rate was 72.7% and 59.1%, respectively. There was no significant difference in 1-year OS rate between the two groups (P>0.05). After induction chemotherapy, the median time for absolute neutrophil count recovery to 0.5×109/L in D-EIAG group and D-CAG group was 14 (10-27) d and 12 (10-26) d, for platelet count recovery to 20×109/L was 15 (11-28) d and 14 (11-24)d, the median red blood cell suspension transfusion volume was 8 (6-12) U and 6 (6-12) U, and the median apheresis platelet transfusion volume was 4 (2-8) U and 3 (2-6) U, respectively. There were no statistically significant differences in comparison of the above indicators between the two groups (P>0.05). The hematological adverse reactions of patients were mainly myelosuppression. Grade III-IV hematological adverse events occurred in both groups (100%), with no increase in the incidence of non-hematological toxicities such as gastrointestinal reactions or liver function damage. CONCLUSION: Decitabine combined with EIAG regimen in the treatment of relapsed/refractory AML and high-risk MDS can improve remission rate, provide an opportunity for subsequent therapies, and have no increase in adverse reactions compared with D-CAG regimen.

Low T3 syndrome as a predictor of poor outcomes in patients with follicular lymphoma.

The aim of this study was to investigate the prognostic value of low T3 syndrome in follicular lymphoma (FL). A total of 221 FL patients with detailed serum thyroid hormone levels and other complete clinical data were enrolled. Baseline features associated with low T3 syndrome were analyzed and balanced by propensity score matching. Univariate and multivariate regression analyses were performed to determine independent risk factors for progression-free survival (PFS) and overall survival (OS). A receiver operating characteristic (ROC) curve was plotted, and the area under the curve (AUC) was calculated to assess the predictive accuracy of FL international prognostic index FLIPI-1/FLIPI-2 and low T3 syndrome. A total of 22 patients (10.0%) had low T3 syndrome at diagnosis, which was associated with poor PFS and OS in the rituximab era. It is an independent prognostic factor for PFS and OS. Low T3 syndrome and FLIPI-1/FLIPI-2 significantly increased the AUC of PFS and OS compared to FLIPI-1/FLIPI-2 alone. Low T3 is a risk factor for POD24. In conclusion, low T3 syndrome may be a good candidate for predicting the prognosis of CLL in future clinical practice. Our study demonstrates that low T3 syndrome is associated with poorer survival outcomes in FL patients.

The novel HLA-A*02:1075 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-A*02:1075 differs from HLA-A*02:07:01:01 by one nucleotide in exon 5.

[Analysis of the causes of residual back pain in the early and late stages after percutaneous vertebral augmentation].

OBJECTIVE: To explore the influencing factors of the residual back pain in patient with osteoporotic vertebral compression fractures(OVCFs) in the early and late stages after percutaneous vertebral augmentation(PVA), and analyze the correlation between these factors and the residual back pain after PVA. METHODS: From March 2018 to December 2019, 312 patients with OVCFs who treated with PVA were collected. According to the inclusion and exclusion criteria, a total of 240 patients were included in this retrospective study. There were 59 males and 181 females, aged from 50 to 95 years old with an average of (76.11±10.72) years old, and 50 cases of fractures located in the thoracic region (T5-T10), 159 cases in the thoracolumbar region (T11-L2), and 31 cases in the lumbar region (L3 and below). The first day after PVA was regarded as the early postoperative period, and the seventh day was regarded as the late postoperative period. According to the visual analogue scale (VAS), the patients were divided into 4 groups:early postoperative pain relief group(group A, VAS≤4 scores), there were 121 patients, including 29 males and 92 females, aged from 50 to 90 years with an average of (75.71±11.00) years;early postoperative pain relief was not an obvious group (group B, VAS >4 scores), there were 119 patients, including 30 males and 89 females, aged from 53 to 95 years with an average of (76.51±10.46) years; late postoperative pain relief group (group C, VAS≤ 4 scores), there were 172 patients, including 42 males and 130 females, aged from 50 to 95 years with an average of (76.20±10.68) years; late postoperative pain relief was not obvious group (group D, VAS>4 scores), there were 68 patients, including 17 males and 51 females, aged from 53 to 94 years old with an average of (75.88±10.91) years old. The age, gender, bone mineral density(BMD), injured vertebral segment, preoperative thoracolumbar fascial condition, surgical methods, single or bilateral puncture, the amount of bone cement injection, anterior vertebral height recovery rate and central vertebral height recovery rate in the 4 groups were analyzed by univariate analysis. The statistically significant factors were put into a Logistic regression to analyze the correlation between these factors and residual back pain after PVA. RESULTS: Univariate analysis showed that the residual back pain in the early stage after PVA was correlated with BMD, preoperative thoracolumbar fascial injury, single or bilateral puncture, the amount of bone cement injection, anterior vertebral height recovery rate and central vertebral height recovery rate(P<0.05). The residual back pain in the late postoperative period was related to BMD, injured vertebral segment, surgical methods, the amount of bone cement injection, anterior vertebral height recovery rate and central vertebral height recovery rate(P<0.05). Multivariate Logistic regression analysis showed that thoracolumbar fascial injury(OR=4.938, P=0.001), single or bilateral puncture(OR=5.073, P=0.002) were positively correlated with the residual back pain in the early stage after PVA(B>0), which were risk factors;the BMD (OR=0.211, P=0.000) and anterior vertebral height recovery rate (OR=0.866, P=0.001) were negatively correlated with the residual back pain in the early stage after PVA(B<0), which were protective factors. In the late stage after PVA, the BMD(OR=0.448, P=0.003), the amount of bone cement injection (OR=0.648, P=0.004) and anterior vertebral height recovery rate (OR=0.820, P=0.000) were negatively correlated with residual back pain(B<0), which were protective factors. CONCLUSION: The decrease of BMD, injury of the thoracolumbar fascia, single or bilateral puncture, poor recovery of anterior vertebral height and insufficient injection of bone cement are closely related to the occurrence of residual back pain after PVA, which affect the relief of residual back pain in the early and late postoperative periods.

USF2 reduces BMP3 expression via transcriptional activation of miR-34a, thus promoting osteogenic differentiation of BMSCs.

INTRODUCTION: Osteoporosis is the most susceptible disease for people over 60. The main cause of osteoporosis is the decreased osteogenic differentiation of mesenchymal stem cells (MSCs). Here we showed that upstream stimulatory factor 2 (USF2)/microRNA-34a (miR-34a)/bone morphogenetic protein 3 (BMP3) axis regulated osteogenic differentiation of BMSCs. MATERIALS AND METHODS: USF2 and miR-34a expression were examined using qPCR. Protein levels of BMP3 and osteogenic markers expression were evaluated using both western blot and qPCR. Activity of ALP was determined by ALP assay kit. Mineralization capacity of hBMSCs was assessed using ARS. Besides, CHIP assay was employed to verify whether USF2 could bind to miR-34a promoter. Finally, RIP assay and dual-luciferase reporter assay were employed to verify whether miR-34a directly bound to BMP3. RESULTS: Our results suggested that miR-34a was upregulated during osteogenic differentiation of BMSCs, and miR-34a overexpression could enhance osteogenic differentiation of BMSCs. USF2 could positively regulate miR-34a expression by interacting with its promoter. USF2 overexpression enhanced osteogenic differentiation of BMSCs, while miR-34a inhibition reversed the effect. Besides, BMP3 was the target of miR-34a. MiR-34a overexpression enhanced osteogenic differentiation of BMSCs, which was abolished by BMP3 overexpression. CONCLUSION: Taken together, USF2 enhanced osteogenic differentiation of BMSCs via downregulating BMP3 by interacting with miR-34a promoter.

Anti-inflammatory effects of kaempferol-3-O-rhamnoside on HSV-1 encephalitis in vivo and in vitro.

BACKGROUND: Herpes simplex virus encephalitis (HSE) is an acute central nervous system infectious disease caused by herpes simplex virus (HSV). Currently, there is no effective treatment for HSE infection, which produces many pro-inflammatory factors. Kaempferol-3-O-rhamnoside (K-3-rh) is a plant flavonoid. This study was investigated the anti-inflammatory effect of K-3-rh on encephalitis induced by HSV-1. METHODS: HSV-1 was co-cultured with VERO cells. Cells were divided into four groups, including the control group, virus group, K-3-rh group, Astragalus polysaccharide (APS) group and dexamethasone group. Flow cytometry were utilized to determine cell apoptosis, respectively. Proteins and mRNAs were estimated by western blot and qRT-PCR, respectively. RESULTS: After viral infection, the cytokines were significantly increased. After K-3-rh intervention, the expression of tumor necrosis factor-α (TNF-α), interleukin-1 beta (IL-1ß), and nitric oxide (NO) in microglia were reduced contrast with those in the virus group, and the expression of interleukin-10 (IL-10) did not change. After viral infection, the apoptotic rate increased significantly, and K-3-rh could inhibit viral-induced apoptosis in the microglial cell line. The induction of microglia apoptosis was achieved by cytochrome c and caspase-9-mediated mitochondrial pathway. Also, the pathological changes of brain tissue in mice of each drug intervention group were alleviated. CONCLUSIONS: In conclusion, K-3-rh had the potential to reduce HSV-1-induced brain injury by reducing the secretion of microglial pro-inflammatory factors, inducing apoptosis of microglia cells, and through cytochrome C and caspase-3 pathway.

Affinity-based protein profiling to reveal targets of puerarin involved in its protective effect on cardiomyocytes.

Natural products are an important source of new drugs. Some of them may be used directly in clinical settings without further structural modification. One of these directly used natural products is puerarin (Pue), which protects cardiomyocytes against oxidative stress and high glucose stress. Although Pue has been used in clinics for many years, its direct binding targets involved in the protection of cardiomyocytes are not yet fully understood. Here, we reported that Pue could prevent cardiomyocytes from apoptosis under H2O2 and high glucose conditions. Based on affinity-based protein profiling methods, we synthesized an active Pue probe (Pue-DA) with a photosensitive crosslinker to initiate a biological orthogonal reaction. Because of the steric hindrance of Pue-DA, two conformational isomers (syn and anti) unequivocally existed in the probe, and these transformed into one isomer when the probe was heated at 60 °C. We confirmed that the alkylation was on the 7-position phenol group of Pue. Mass spectroscopy revealed that Pue-DA can bind with three proteins, namely CHAF1B, UBE2C, and UBE2T. Finally, cellular thermal shift assay showed that Pue has the ability to stabilize CHAF1B stabilization. The knock-down of CHAF1B reduced the protective effect of Pue on cardiomyocytes. In conclusion, Pue protects cardiomyocytes from apoptosis through binding with CHAF1B.

Factors Defining the Development of Severe Illness in Patients with COVID-19: A Retrospective Study.

OBJECTIVE: Early triage of patients with coronavirus disease 2019 (COVID-19) is pivotal in managing the disease. However, studies on the clinical risk score system of the risk factors for the development of severe disease are limited. Hence, we conducted a clinical risk score system for severe illness, which might optimize appropriate treatment strategies. METHODS: We conducted a retrospective, single-center study at the JinYinTan Hospital from January 24, 2020 to March 31, 2020. We evaluated the demographic, clinical, and laboratory data and performed a 10-fold cross-validation to split the data into a training set and validation set. We then screened the prognostic factors for severe illness using the least absolute shrinkage and selection operator (LASSO) and logistic regression, and finally conducted a risk score to estimate the probability of severe illness in the training set. Data from the validation set were used to validate the score. RESULTS: A total of 295 patients were included. From 49 potential risk factors, 3 variables were measured as the risk score: neutrophil to lymphocyte ratio ( OR, 1.27; 95% CI, 1.15-1.39), albumin ( OR, 0.76; 95% CI, 0.70-0.83), and chest computed tomography abnormalities ( OR, 2.01; 95% CI, 1.41-2.86) and the AUC of the validation cohort was 0.822 (95% CI, 0.7667-0.8776). CONCLUSION: This report may help define the potential of developing severe illness in patients with COVID-19 at an early stage, which might be related to the neutrophil to lymphocyte ratio, albumin, and chest computed tomography abnormalities.

Artesunate promotes osteoblast differentiation through miR-34a/DKK1 axis.

BACKGROUND: Osteoporosis is characterised by impairment of microarchitecture and bone mass. Therapeutic strategy promoting osteoblast differentiation is considered as a promising direction for the treatment of osteoporosis. Artesunate (ART) is related to osteoporosis, but the relationship between ART and osteogenic differentiation is still unknown. METHODS: Cells proliferation were measured by MTT, ALP activity assay and Alizarin Red S staining were used to assess osteogenic differentiation of hBMSCs. Western blotting and qRT-PCR were applied for measuring expression of protein and mRNA, respectively. The relationship between miR-34a and Dickkopf-1 (DKK1) was detected by dual luciferase reporter assay. RESULTS: The expression of osteoblasts differentiation related proteins (Runx2, OCN, and OPN) were significantly increased by ART. And ART accelerates the osteoblasts differentiation of hBMSCs through promoting Wnt signaling pathway by DKK1 inhibition. Significant higher expression of miR-34a and lower expression of DKK1 could be induced by ART. We firstly proved that miR-34a could bind DKK1 specifically. CONCLUSION: ART could promote osteoblast differentiation through miR-34a/DKK1/Wnt pathway. Therefore, our findings may provide a new thought for the treatment of osteoporosis by ART through osteoblast differentiation promotion.

[Mechanism action of Chinese herbal compound and target network pharmacology of Yougui (YG) pill for the treatment of osteoporosis].

OBJECTIVE: To explore compounds, targets and mechanism of Yougui (YG) pill in treating osteoporosis based on systemic pharmacology of traditional Chinese medicine. METHODS: The known effective Chinese herbal compound of YG pill was searched from traditional Chinese medicine integrated database(TCMID). Bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine (BATMAN-TCM) was used to predict target of components;DisGeNET and artificial literature reading were used to obtain targets of osteoporosis and bone remodeling;Cytoscape 3.7.1 software and its plug-ins BiN-GO and ClueGO were used to enrich the GO annotation and pathwaysof the related targets, and validation of the predicted target of YG pill were validated by 87 differentially expressed proteins in postmenopausal osteoporosis and postmenopausal osteoporosis disease models in postmenopausal patients with normal bone mass from the previous serum proteomics data. RESULTS: Totally 392 compounds were retrieved from YG pill, including 83 sovereign drugs (monkshood, cinnamon, deerhorn gelatin), 127 ministerial drugs (prepared rehmannia root, dogwood, wolfberry fruit and Chinese yam) and 182 supplementary drugs (cuscuta chinensis, eucommia ulmoides and Chinese angelica). Among them, there were 4 same compounds between sovereign drug and ministerial drug, 1 same compound between sovereign drug and supplementary drug, and 14 same compounds between ministerial drug and supplementary drug. Totally 2 112 trusted targets were identified, included 775 sovereign drugs, 1 483 ministerial drugs and 1 491 supplementary drugs;227 targets were selected from YG pill for treating osteoporosis, which participate in nearly 20 process of metabolic process, cell differentiation and biology, and data mining revealed that the process involved bone remodeling and bone mineralization. Acting site of cell mainly inclded 9 kinds of cell which had 13 molecular function. Results of KEGG metabolic pathway enrichment analysis showed 137 signal passages were obviously enriched. Among them, classical osteoclast differentiation signal passages and relative estrogen regulates signaling pathways of menopause were widely distributed in 27 signal passages. Sixtargets were screened by target validation, such as AGT, FGA, APOE, DKK3, P4HB and RAB7A. CONCLUSION: The characteristics of multi-targets and multi-pathways of YG pill for the treatment of osteoporosis were clarified, which provided a clear direction for the in-depth research. The pharmacodynamic components of YG pill include 36 compounds, and their main action targets include FGA, AGT, APOE, DKK3, P4HB and RAB7A.

[Analysis of the related factors of fever of unknown causes before operation of intertrochanteric fracture in the elderly].

OBJECTIVE: To explore the related factors of fever of unknown causes before operation of intertrochanteric fracture of femur in the elderly. METHODS: From August 2015 to August 2018, 156 cases of intertrochanteric fracture of femur were treated by intramedullary nail fixation. According to the preoperative measurement, whether there was fever was divided into fever group and non fever group. There were 80 cases in fever group, 26 males and 54 females, aged 60 to 93 (75.063±13.082) years; 76 cases in non fever group, 39 males and 37 females, aged 60 to 96 (74.763±13.692) years. All patients' sex, age, basic diseases, WBC, NE%, Hb, CRP, D dimer and ALB were observed for single factor analysisand multi factor analysis. RESULTS: Single factor analysis showed that gender, Hb, CRP had influence on preoperative fever, but other indexes had no influence; multi factor analysis showed that gender and CRP might have influence on fever. The probability of fever increased by 1.2%(P=0.050) for every increase of CRP, and there was no significant difference between female and male in the probability of fever (P=0.061). CONCLUSION: CRP is an independent risk factor of preoperative fever in the elderly patients with intertrochanteric fracture of femur. The abnormality of CRP indicates that patients are more likely to have preoperative fever. Perioperative management should pay attention to the monitoring of CRP and early intervention.

In vitro Transport Ability of ABCC2 (G1249A) Polymorphic Variant Towards Anticancer Drugs.

OBJECTIVE: Multidrug resistance-associated protein 2 (MRP2), encoded by ABCC2 gene, is involved in the efflux of certain anticancer drugs. Here we observed whether the ABCC2 (G1249A) polymorphism impacts the transport abilities of MRP2-dependent paclitaxel, docetaxel, and doxorubicin in recombinant LLC-PK1 cell lines. METHODS: LLC-PK1 cell lines transfected with ABCC2 1249G wild-type and ABCC2 1249A variant alleles were used to evaluate the sensitivity, intracellular accumulation, and transmembrane transport of paclitaxel, docetaxel, and doxorubicin. RESULTS: The recombinant ABCC2 1249A variant cell line showed higher IC50 values for paclitaxel and doxorubicin than ABCC2 1249G wild-type cell system (p<0.01). Intracellular accumulations of paclitaxel and doxorubicin in cells transfected with ABCC2 1249A variant allele were significantly decreased compared to cells transfected with ABCC2 1249G wild-type allele (p<0.01). The efflux ratios of paclitaxel and doxorubicin across ABCC2 1249A cell line were significantly increased compared with ABCC2 1249G cell system (p<0.01). However, ABCC2 (G1249A) polymorphism had no effect on the transport activity of MRP2-mediated docetaxel. CONCLUSION: Our results indicate that ABCC2 (G1249A) polymorphism affects the transport activities of MRP2-dependent paclitaxel and doxorubicin, resulting in greater efflux of these anticancer drugs.

[Analysis of Related Factors Affecting the Nosopoiesis of Childhood Acute Leukemia].

OBJECTIVE: To analyze the related factors affecting the nosopoiesis of childhood acute leukemia from the perspective of indoor environmental exposure, behavior and lifestyle. METHODS: The clinical data of 64 children with acute leukemia were retrospectively analyzed, and 50 healthy children were selected as the control group during the same period. The basic data of children, indoor environment, behavior and lifestyle of parents in 2 groups were recorded. Univariate and multivariate logistic regression analysis were used to analyze the related factors affecting the incidence of childhood acute leukemia, and the OR (95%CI) value was calculated. RESULTS: The unvariate analysis showed that the daily wine-drinking rate of father and pesticide use rate in acute leukemia group were significantly higher than those in control group (P<0.05). Multivariate Logistic regression analysis showed that indoor ventilation during summer sleep of children (OR=0.35, 95%CI: 0.14-0.88) and contact with other children before 3 years old (OR=0.34, 95%CI: 0.18-0.65) were protective factors for provention of childhood acute leukemia (P<0.05). Mothers had a history of exposure to chemical substances (OR=3.68, 95%CI: 1.64-8.27), and children had a history of exposure to chemical substances (OR=3.84, 95%CI: 1.64-9.01), family had internal decoration history after child birth (OR = 1.38, 95%CI: 1.05-1.81) and family uses of pesticides (OR=2.17, 95%CI: 1.08-4.36), all these factors were independent risk factors for acute leukemia (P<0.05). CONCLUSION: Indoor environmental exposure, behavior and lifestyle of children and parents may be closely related with the nosopoiesis of childhood acute leukemia.

The Mediating Effect of Body Dissatisfaction in Association between Obesity and Dietary Behavior Changes for Weight Loss in Chinese Children.

OBJECTIVE: The aim of this study was to analyze the mediating effect of body dissatisfaction in correlation between obesity and dietary behavior changes for weight loss (DBCWL). METHODS: A total of 680 primary and middle school students were included in this study. Their body height, weight, and waistline were effectively measured, and they were also evaluated to assess their body dissatisfaction, perception of dietary behaviors, and DBCWL. The correlation among these factors was analyzed using mediating effect models. RESULTS: The prevalence of overweight/obesity and abdominal obesity was significantly higher in males than in females (P < 0.05). Overweight/obesity, abdominal obesity, and body dissatisfaction significantly increased the risk for DBCWL (OR = 2.57, 2.77, and 1.95, respectively). Overweight/obesity and abdominal obesity significantly increased the risk for body dissatisfaction (OR = 6.00 and 4.70, respectively). Significant mediating effects of body dissatisfaction were observed in correlation between overweight/obesity and DBCWL and between abdominal obesity and DBCWL (OR = 2.20 and 1.92, respectively; P < 0.05), and the proportions of mediating effects among the total effects were 48.89% and 46.60%, respectively. CONCLUSION: Body dissatisfaction might play an important mediating effect in association between DBCWL and obesity, which indicates that guiding children to correctly recognize their body might be more conducive than promoting obese children toward weight loss through dietary behavior changes.

Frequency of Diarrheagenic Virulence Genes and Characteristics in Escherichia coli Isolates from Pigs with Diarrhea in China.

Intestinal pathogenic Escherichia coli (InPEC) is a leading cause of postweaning diarrhea (PWD) in pigs. Here, a total of 455 E. coli strains were isolated from small intestinal content or feces from pigs with PWD in 56 large-scale (>500 sows; 10,000 animals per year) swine farms between 2014 and 2016. The frequency of occurrence of selected virulence factors for InPEC pathotypes was detected in 455 isolates by real-time PCR. Sequence types (STs), pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility profiles of 171 E. coli isolates from 56 swine farms were further determined. The heat-labile enterotoxin (LT) was the most common (61.76%), followed by heat-stable enterotoxin (STb) (33.19%), stx2e (21.54%), STa (15.00%), eae (8.98%), cnf2 (5.71%), stx2 (5.71%), F18 (3.25%), and F4 (2.25%) with rates varying by geographic area and year of isolation. Notably, hybrids of E. coli isolates were potentially more virulent, as some InPEC hybrids (virotype F18:LT:eae:stx2e) can rapidly cause cell death in vitro. Genotypic analysis revealed that the most prominent genotype was ST10 (12.87%). The PFGE patterns were heterogeneous but were not ST or virotype related. A total of 94.15% of isolates were multidrug-resistant, with average resistance rates ranging from 90.05% for nalidixic acid to 2.34% for meropenem. Our investigation contributes to establishing the etiology of diarrhea and developing intervention strategies against E. coli-associated diarrheal disease in the future.