Chemical constituents from whole herb of Hedyotis scandens / 中国中药杂志

This study aimed to investigate the chemical constituents in the water extract of the whole herb of Hedyotis scandens by silica gel, ODS, and MCI column chromatographies together with preparative high-performance liquid chromatography(HPLC). The structures of isolated constituents were identified by NMR, HR-ESI-MS, etc. Thirteen compounds were isolated and identified as methyl 4-benzoyloxy-3-methoxybenzeneacetate(1), 4-benzoyloxy-3-methoxybenzeneacetic acid(2), 3-(4-hydroxy-3-methoxyphenyl)-propanoic acid(3), salicylic acid(4), 3-hydroxy-4-methoxypyridine(5), syringic acid(6), hydroxycinnamic acid(7),(R)-6-methyl-4,6-bis(4-methylpent-3-enyl)cyclohexa-1,3-dienecarbaldehyde(8), 1,2-bis(4-hydroxy-3-methoxyphenyl)-1,3-propanediol(9), 1H-indole-3-carboxaldehyde(10), isoscopoletin(11), syringaresinol(12), and pinoresinol(13). Among them, compounds 1 and 2 were new phenolic acid compounds, compounds 3-5, 8-11, and 13 were isolated from this genus for the first time, and compounds 6, 7, and 12 were obtained from H. scandens for the first time. The activity test showed that compounds 1 and 10 had a certain inhibitory effect on Mycobacterium smegmatis, with MIC_(50) values of 58.5 and 33.3 μg·mL~(-1), respectively.

Research progress on tumor treating fields combined with radiation therapy in the treatment of glioblastoma / 中华放射肿瘤学杂志

For patients with newly diagnosed glioblastoma, tumor treating fields (TTF) combined with temozolomide after radiation therapy is currently one of the standard therapeutic regimens. Recently, TTF has been increasingly applied in combination with radiation therapy since it can delay tumor DNA repair and increase DNA replication stress. The efficacy of TTF has been proven in clinical studies. However, no consensus has been reached regarding the theoretical basis, radiation dose, actual clinical operation, patients' benefit and safety, which remain controversial. In this article, research progress on these topics was reviewed.

Mechanosensitive ion channel Piezo1 induces senescence of atrial fibroblasts by activating β-catenin / 中国药理学通报

Aim To observe whether the mechanosensitive ion channel Piezo1 was involved in the senescence of atrial fibroblasts by activating β-catenin based on our previous study which found marked increase of Piezo1 mRNA in senescent atrial fibroblasts. Methods Primary mouse atrial fibroblasts (MAFs) were isolated from male C57BL/6 mice (3-4 weeks) by enzyme digestion, and tert-butyl hydroperoxide (TBHP) was used to induce the senescence of cells. The ratio of senescent cells was detected by senescence-associated β-galactosidase (SA-β-Gal) staining. The protein levels of Piezo1, β-catenin/p-β-catenin, senescence-associated proteins p53 and p21 in the cells treated with TBHP (100 μmol · L

Oral administration of Bifidobacterium breve improves anti-angiogenic drugs-derived oral mucosal wound healing impairment via upregulation of interleukin-10 / 国际口腔科学杂志·英文版

Recent studies have suggested that long-term application of anti-angiogenic drugs may impair oral mucosal wound healing. This study investigated the effect of sunitinib on oral mucosal healing impairment in mice and the therapeutic potential of Bifidobacterium breve (B. breve). A mouse hard palate mucosal defect model was used to investigate the influence of sunitinib and/or zoledronate on wound healing. The volume and density of the bone under the mucosal defect were assessed by micro-computed tomography (micro-CT). Inflammatory factors were detected by protein microarray analysis and enzyme-linked immunosorbent assay (ELISA). The senescence and biological functions were tested in oral mucosal stem cells (OMSCs) treated with sunitinib. Ligated loop experiments were used to investigate the effect of oral B. breve. Neutralizing antibody for interleukin-10 (IL-10) was used to prove the critical role of IL-10 in the pro-healing process derived from B. breve. Results showed that sunitinib caused oral mucosal wound healing impairment in mice. In vitro, sunitinib induced cellular senescence in OMSCs and affected biological functions such as proliferation, migration, and differentiation. Oral administration of B. breve reduced oral mucosal inflammation and promoted wound healing via intestinal dendritic cells (DCs)-derived IL-10. IL-10 reversed cellular senescence caused by sunitinib in OMSCs, and IL-10 neutralizing antibody blocked the ameliorative effect of B. breve on oral mucosal wound healing under sunitinib treatment conditions. In conclusion, sunitinib induces cellular senescence in OMSCs and causes oral mucosal wound healing impairment and oral administration of B. breve could improve wound healing impairment via intestinal DCs-derived IL-10.

Establishment and preliminary application of quantitative real-time PCR assay for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA / 中华预防医学杂志

Objective: To establish a rapid and specific quantitative real-time PCR (qPCR) method for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA (SgN) in patients with COVID-19 or environmental samples. Methods: The qPCR assay was established by designing specific primers and TaqMan probe based on the SARS-CoV-2 genomic sequence in Global Initiative of Sharing All Influenza Data (GISAID) database. The reaction conditions were optimized by using different annealing temperature, different primers and probe concentrations and the standard curve was established. Further, the specificity, sensitivity and repeatability were also assessed. The established SgN and genomic RNA (gRNA) qPCR assays were both applied to detect 21 environmental samples and 351 clinical samples containing 48 recovered patients. In the specimens with both positive gRNA and positive SgN, 25 specimens were inoculated on cells. Results: The primers and probes of SgN had good specificity for SARS-CoV-2. The minimum detection limit of the preliminarily established qPCR detection method for SgN was 1.5×102 copies/ml, with a coefficient of variation less than 1%. The positive rate of gRNA in 372 samples was 97.04% (361/372). The positive rates of SgN in positive environmental samples and positive clinical samples were 36.84% (7/19) and 49.42% (169/342), respectively. The positive rate and copy number of SgN in Wild strain were lower than those of SARS-CoV-2 Delta strain. Among the 25 SgN positive samples, 12 samples within 5 days of sampling time were all isolated with virus; 13 samples sampled for more than 12 days had no cytopathic effect. Conclusion: A qPCR method for the detection of SARS-CoV-2 SgN has been successfully established. The sensitivity, specificity and repeatability of this method are good.

Piezol involved in electrical remodeling in atrial myocytes induced by high hydrostatic pressure through Src kinase / 中国药理学通报

Aim To investigate the role of mechano- sensitive ion channel Piezol in regulating electrical re-modeling of atrial myocytes induced by hypertension and to further explore the potential mechanisms.Methods Spontaneously hypertensive rats ( SHR ) aged 30 - 32 weeks treated with or without valsartan (30 mg • kg 1 • d 1 ) were used.Wistar rats were used as control.Western blot was used to detect the protein expression of Piezol , Src and Cavl.2 in atrial appendages of rats and in atrial myocytes ( HL-1 cells) exposed to different levels of high hydrostatic pressure (20 and 40 mmHg) , Piezol inhibitor (GsmTx4) and agonist ( Yodal ) in vitro.Whole-cell patch clamp technique was employed to detect L-tvpe calcium current (ICa, ) and action potential duration ( APD) of atrial myocytes.Results Compared with Wistar rats in control group, the protein expressions of Piezol and Src significantly increased and the expression of Cavl.2 decreased in SHR group (P < 0.05 ), while the a- bove changes could he reversed in SHR treated with valsartan( P < 0.05 ) .Meanwhile, higher hydrostatic pressure (40 mniHg) could increase the expressions of Piezol and Src in HL-1 cells( P <0.05) and decrease the protein expression of Cavl.2 (P <0.05 ) , accompanied by a shortened APD and a decreased ICa,.GsmTx4 could significantly reverse the above changes.In addition, Piezol agonist Yodal could simulate electrical remodeling and related signal molecule changes in atrial myocytes induced by the high hydrostatic pressure.Conclusions Mechanosensitive ion channel Piezol participates in electrical remodeling induced by hypertension via activating Src kinase signaling pathway and then leading to the decrease of ICa ,.

Role of p300 in susceptibility of atrial Hbrillation in aged mice / 中国药理学通报

Aim To explore the role of cotranscriptional activator p300 in regulating the electrical remodeling of atrial myocytes in aging mouse, which resulted in atrial fibrillation. Methods The left atrial appendage tissues of 5 , 13 and 18monthold C57BL/6 mice were collected respectively. Western blot was used to detect the protein expression levels of p300, L type calcium channel (Cavl. 2) and aging related protein p53/p21. Acute enzymatic hydrolysis was used to isolate single atrial myocytes, and the wholecell patchclamp technique was used to detect the Ltype calcium current (I

Value of regional cerebral oxygen saturation and anesthesia depth in predicting postoperative cognitive dysfunction in patients with non-macrovascular surgery / 中国医师杂志

Objective:To explore the value of regional cerebral oxygen saturation (rScO 2) and anesthesia depth monitoring in predicting postoperative cognitive dysfunction (POCD) in patients with non-macrovascular surgery. Methods:A retrospective analysis of 147 patients with non-macrovascular surgery under general anesthesia admitted to the First Affiliated Hospital of Xi'an Jiaotong University from August 2017 to June 2019 was performed and divided into the POCD group ( n=37) and the non-POCD group ( n=110) according to the presence/absence of postoperative POCD. The changes of bispectral index (BIS) and rScO 2 in patients before anesthesia induction (T 0), endotracheal intubation (T 1), 2 hours after operation (T 2), after operation (T 3), and at extubation (T 4) were recorded, and the predictive value for the occurrence of POCD was analyzed by receiver operating characteristic (ROC) curve. Results:There was no statistically significant difference in anesthesia time, operation time and operation type between the two groups ( P>0.05). There was no significant difference in BIS and rScO 2 levels between the two groups at T 0, T 1 and T 4 ( P>0.05). BIS and rScO 2 levels in the POCD group at T 2 and T 3 were lower than those in the non-POCD ( P<0.05). Both BIS and rScO 2 of the two groups reached the lowest value at T 2, and the reduction rate of rScO 2 in the POCD group was higher than that in the non-POCD group [(31.84±3.27)% vs (14.81±2.52)%, P<0.05]. The ROC curve of BIS-T 2, rScO 2-T 2, BIS-T 3, rScO 2-T 3, rScO 2 reduction from the baseline value to predict POCD in patients with non-macrovascular surgery was plotted, and the AUCs were 0.514, 0.617, 0.505, 0.633, 0.724, respectively. The highest AUC value of 0.808 was found for combined detection at T 2 (rScO 2 and BIS). Conclusions:The combined detection of intraoperative regional cerebral oxygen saturation and anesthesia depth monitoring is of good clinical application value in predicting postoperative cognitive dysfunction in patients with non-macrovascular surgery.

Salubrinal increases the apoptosis of oral cancer cells by inhibiting radiation-induced activation of NF-Κb / 中华放射肿瘤学杂志

Objective@#To explore the mechanism of the role of Salubrinal in regulating the radiation-induced apoptosis of oral cancer cells.@*Methods@#Radioresistant KBR cell line was constructed (4 Gy per fraction, every 7-10 d for 4 times). The radiosensitivity of oral cancer cells after Salubrinal pretreatment was measured by colony formation assay. The expression levels of NF-κB-HIF-1α signaling pathway and apoptosis biomarker cleaved PARP in oral cancer cells were measured by Western blot. The apoptosis rate was detected by Annexin V, PI staining and flow cytometry.@*Results@#Colony formation assay demonstrated that Salubrinal increased the radiosensitivity of oral cancer cells. The radiosensitization ratios of KB and KBR cells were 1.19 and 1.24. Western blot revealed that the activation of NF-κB-HIF-1α was time-dependent in the radiation-induced oral cancer cells, whereas Salubrinal inhibited the radiation-induced abnormal activation. In addition, Salubrinal increased the expression of apoptosis biomarker cleaved PARP and apoptosis index in radiation-induced oral cancer cells, whereas TNF-α, an activator of NF-κB, reversed the effect, suggesting that Salubrinal increased the apoptosis of radiation-induced oral cancer cells by suppressing the activation of NF-κB. Pretreatment of NF-κB inhibitor Bay11-7082 also increased the cell apoptosis. The expression levels of cleaved PARP of KB and KBR cell lines in the Bay11-7082+ IR group were 2.67±0.26 and 1.91±0.17, significantly higher compared with 2.1±0.16 and 1.44±0.15 in the IR group (both P<0.05).@*Conclusion@#Salubrinal can aggravate the apoptosis of radiation-induced oral cancer cells by inhibiting the radiation-induced activation of NF-κB, thereby regulating the radiosensitivity of oral cancer cells.

Salubrinal increases the apoptosis of oral cancer cells by inhibiting radiation-induced activation of NF-κB / 中华放射肿瘤学杂志

Objective To explore the mechanism of the role of Salubrinal in regulating the radiation-induced apoptosis of oral cancer cells.Methods Radioresistant KBR cell line was constructed (4 Gy per fraction,every 7-10 d for 4 times).The radiosensitivity of oral cancer cells after Salubrinal pretreatment was measured by colony formation assay.The expression levels of NF-κB-HIF-1o signaling pathway and apoptosis biomarker cleaved PARP in oral cancer cells were measured by Western blot.The apoptosis rate was detected by Annexin V,PI staining and flow cytometry.Results Colony formation assay demonstrated that Salubrinal increased the radiosensitivity of oral cancer cells.The radiosensitization ratios of KB and KBR cells were 1.19 and 1.24.Western blot revealed that the activation of NF-κB-HIF-1α was time-dependent in the radiation-induced oral cancer cells,whereas Salubrinal inhibited the radiation-induced abnormal activation.In addition,Salubrinal increased the expression of apoptosis biomarker cleaved PARP and apoptosis index in radiation-induced oral cancer cells,whereas TNF-α,an activator of NF-κB,reversed the effect,suggesting that Salubrinal increased the apoptosis of radiation-induced oral cancer cells by suppressing the activation of NF-κB.Pretreatment of NF-κB inhibitor Bay1 1-7082 also increased the cell apoptosis.The expression levels of cleaved PARP of KB and KBR cell lines in the Bay11-7082+IR group were 2.67±0.26 and 1.91±0.17,significantly higher compared with 2.1±0.16 and 1.44±0.15 in the IR group (both P＜0.05).Conclusion Salubrinal can aggravate the apoptosis of radiation-induced oral cancer cells by inhibiting the radiation-induced activation of NF-κB,thereby regulating the radiosensitivity of oral cancer cells.

Pyroptosis induced by different Enteroviruses infection in SH-SY5Y cell / 中华实验和临床病毒学杂志

Objective@#To investigate the pyroptosis induced by different enteroviruses in human neuroblastoma cells SH-SY5Y and the differences among them.@*Methods@#SH-SY5Y cells were infected with nine strains of enterovirus respectively, including enterovirus A71 (EV-A71), Coxsackievirus A (CA), Coxsackievirus B (CB), Echovirus (Echo). The cellular morphology of infected and control groups were observed and activity of Caspase-1 of infected and control groups were detected by flow cytometry at 48 h post infection.@*Results@#The activity of Caspase-1 induced by EV-A71 was higher than control (P<0.001), and the activity of Caspase-1 induced by EV-A71 isolated from severe case was significantly higher than that induced by EV-A71 isolated from common case (P<0.001). The activity of Caspase-1 induced by CA was at a lower level. The activity of Caspase-1 induced by CB and Echo were both significantly higher than that induced by EV-A71 and control (P<0.001). Cytopathic effects (CPE) were found to be related with the activity of Caspase-1.@*Conclusions@#EV-A71, CB and Echo all could induce pyroptosis mediated by Caspase-1 in SH-SY5Y cells, but the ability of CA to induce pyroptosis was at a lower level, which may provide evidences for further study on mechanism of neuropathy caused by enterovirus.

Effect of Traditional Chinese Medicine and Western Medicine Therapy on Gestational Diabetes Mellitus / 中国实验方剂学杂志

In recent years, as the level of economic life has improved, the incidence of gestational diabetes mellitus has increased year by year. Gestational diabetes mellitus (GDM) has been a serious threat to maternal and newborn health. The pathogenesis of gestational diabetes is not very clear, and may be closely associated with insulin resistance, genetic susceptibility, inflammatory response, metabolic disorders. According to the gestational diabetes diagnostic standard,24-28 weeks pregnant women keep an empty stomach over 8 h, taken 75 g oral glucose directly, and then receive the oral glucose tolerance test. GDM is diagnosed as fasting blood-glucose> 5.1 mmol · L-1,1-hour postprandial blood glucose>10.0 mmol · L-1,and 2-hour postprandial blood glucose>8.5 mmol · L-1. Western medicine treatment is mainly based on diet, exercise, drugs, education, monitoring and insulin therapy according to blood glucose. Meanwhile, GDM is a type of diabetes in traditional Chinese medicine. GDM is prevented and treated with diets and traditional method sports and Chinese herbs. Therefore, integrated Chinese and western medicine therapy can maximize the curative effect, reduce the incidence of gestational diabetes mellitus, and effectively improve the adverse outcome and prognosis of patients with gestational diabetes mellitus from mother to child.

Complete chloroplast genome of Allium chinense: comparative genomic and phylogenetic analysis / 药学学报

italic>Allium chinense belongs to the genus Alliums of the lily family. It can be used both as medicine and food. To date, the phylogenetic relationship of Allium species have not be resolved completely. Furthermore, there has been a lack of DNA barcode to distinguish closely related species. In this study, the complete chloroplast genome of A. chinense was obtained using next generation DNA sequencing and bioinformatic analysis, and compared with that from other Allium species. The genome is a circular molecule of 152 525 bp with a typical quadripartite structure. Genome annotation identified a total of 116 genes, including 81 protein-coding genes, 31 tRNA genes, and 4 rRNA genes. Analyses of sequences from six Allium species showed that the most diverse regions are found in the protein coding regions such as ndhA and ycf1 genes, and in the intergenic regions, such as ps16-trnQ, trnT-trnF, ndhF-rpl32, rpl32-trnL and rpl16-rps3. A phylogenetic tree was constructed using 58 protein coding sequences from 53 species. All branches showed strong support with bootstrap scores reaching 66%-100%, except those for the Lilium and Paris. Our results suggest that the completed chloroplast genome could solve the classification problems of these species. Using EcoPrimer software, we identified seven markers from the chloroplast genomes, which can be used to differentiate congeneric species. In summary, we have sequenced the complete chloroplast genome of A. chinense, carried out phylogenetic analysis and identified a series of genus specific DNA barcode sequences. The results have laid the foundation for the systematical determination of the phylogenetic relationship of Allium species and the differentiation of species using the genus specific primers.

Extraction and identification of human stem cells from the apical papilla derived-exosomes / 上海交通大学学报（医学版）

Objective: To obtain and identify the exosomes derived from human stem cells from the apical papilla (hSCAPs). Methods: hSCAPs were cultured by modified tissue adherence method and the phenotypes were analyzed with stem cell surface markers CD105, CD45, CD44, CD31, CD34 and CD29. The capability of multi-differentiation in hSCAPs was identified by osteogenic and adipogenic differentiation in vitro. Exosomes were isolated from hSCAPs culture supernatants using gradient centrifugation methods. The size of vesicle was assessed by nanoparticle size analyzer. The morphology of exosomes was observed by transmission electronic microscope (TEM), and the expression of exosome molecular markers CD81, CD9, CD63 and TSG101 was analyzed by Western blotting. Results: hSCAPs were positive for the mesenchyme stem cell markers, including CD105, CD44 and CD29 and negative for the hematopoietic markers CD45, CD31 and CD34. hSCAPs could differentiate into osteoblasts and adipocytes. hSCAPs secreted microvesicles which exhibited round vesicle structure with an intact membrane observed by the TEM. The results of nanoparticle size analyzer measurement showed that the diameters of vesicles were ranged from 30 to 100 nm, which were consistent with the results by TEM. Microvesicles could express the molecular markers for exosomes, i.e. CD81, CD9, CD63 and TSG101. Conclusion: The microvesicles were successfully isolated from hSCAPs and identified as exosomes.

Semaphorin 3A-stimulated bone marrow mesenchymal stem cells sheets promotes osteogenesis of type 2 diabetic rat / 中华口腔医学杂志

Objective@#To evaluate the effect of semaphorin 3A (Sema3A) pre-treated bone marrow mesenchymal stem cells (BMSC) sheets on new bone formation in type 2 diabetes mellitus rats.@*Methods@#Type 2 diabetes mellitus (T2DM) were induced by injection of streptozotocin, and the BMSC were isolated, controlled, identified and induced into cell sheets. Fifteen T2DM rats were randomly divided into control, sheets and Sema3A-sheets group and the calvarial critical size defect (CSD) model of rats were established. The defect zone of rats from control group were implanted with bone powder. The defect zone of rats from sheets group were implanted with bone powder and BMSC sheets. The defect zone of rats from Sema3A-sheets group were implanted with bone powder and BMSC sheets pretreated with 1.0 mg/L Sema3A. After 8 weeks, the bone samples were harvested and analyzed by micro-CT scanning, HE staining for the evaluation of new bone formation, and the immunohistochemical analysis for the expression of osteogenesis-related proteins including type Ⅰ collagen (COL- Ⅰ ), bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN).@*Results@#The BMSC were isolated and cultured, and oil red O and Alizarin red S staining proved the multi-potential differentiation. Eight weeks after the establishment of calvarial CSD model, Sema3A-sheet group showed the most abundant new bone formation (0.516±0.070), with increased bone volume fraction, namely bone volume/tissue volume (BV/TV) compared with sheets group (0.319±0.050) and control group (0.224±0.037) (P<0.05), and the sheets group showed increased BV/TV compared with control group (P<0.05). While trabecular thickness (Tb.Th) control group showed no difference in three groups (P>0.05). HE staining also confirmed that Sema3A-sheets group showed the most new bone formation. Sheet group (0.174±0.051) compared showed difference with control group (0.099±0.033) (P< 0.05), and Sema3A-sheet group (0.421±0.069) showed increased bone formation compared with sheet group and control group (P<0.05). Immunohistochemistry showed that BMSC sheet increased the expression of osteogenesis-related proteins including COL-Ⅰ, BMP-2 and OCN, while Sema3A pretreatment showed more obvious increase of the expression of COL-Ⅰ and OCN.@*Conclusions@#The combined implantation of bone powder and Sema3A stimulated BMSC sheets significantly increased bone regeneration in vivo. Therefore, Sema3A pre-treated BMSC sheets transplantation provides a new strategy for restoring bone defect in T2DM.

Mechanism of endoplasmic reticulum stress pathway inhibitor salubrinal enhancing the apoptosis of head and neck squamous carcinoma cells / 中华放射肿瘤学杂志

Objective To explore the mechanism underlying the effect of endoplasmic reticulum stress pathway inhibitor Salubrinal on enhancing the apoptosis of head and neck squamous carcinoma cells.Methods Three types of head and neck squamous carcinoma cell lines (KB,Fadu,Detroit562) were divided into the control,Salburinal (sal),irradiation (IR) and sal combined with IR (IR+sal) groups.The expression levels of p-ATM/ATM,DNA-PK and cleaved Caspase-3 were quantitatively measured.The cell apoptosis rate was detected among four groups.The effect of Salburinal on cell viability was evaluated by MTT assay.Results Compared with the IR group,the expression level of p-ATM/ATM (t =3.5,8.43 and 9.42,all P＜0.05) was significantly up-regulated,whereas that of DNA-PK (t =9.19,17.44,16.67,all P＜ 0.05) was considerably down-regulated in the IR+sal group.The expression level of cleaved Caspase-3 in the IR+sal group was significantly higher compared with those in the other three groups (t=6.79,9.76 and 9.7g,all P＜0.05).Compared with the IR group,the cell apoptosis rate was significantly enhanced in the IR +sal group (t=5.67,6.95 and 7.28,all P＜0.05).Salubrinal exerted an effect upon the apoptosis of three cell lines in a concentration-and time-dependent manner.Conclusions As an endoplasmic reticulum stress pathway inhibitor,Salubrinal can enhance the apoptosis rate of head and neck squamous carcinoma cells.The underlying mechanism is probably correlated with irradiation-induced DNA double strand injury,suppressing the repairing of DNA damage and thereby increasing the apoptosis of tumor cells.

Stable G6PD knockdown inhibits the migration of renal cell carcinoma cells / 医学研究生学报

Objective Clear cell renal cell carcinoma (ccRCC) accounts for more than 80% of malignant kidney tumors and its pathogenesis has not been elucidated. Our previous studies showed a positive correlation of Glucose-6-phosphate dehydrogenase (G6PD) with the development, progression and poor prognosis of ccRCC. In this study, we first established a G6PD defect ccRCC stable cell line, detected the influence of G6PD knockdown on ccRCC migration, and provided a cell model for further studies on the functional and molecular mechanisms of G6PD in ccRCC.Methods Using the OligoEngine RNAi software, we designed siRNA targeting the human G6PD gene 3′ non-coding region and negative control siRNA sequences, inserted the double-stranded siRNA into the pSR-GFP/Neo expression vector through Bgl Ⅱ and Hind Ⅲ enzyme loci, and constructed Caki-1-G6PD siRNA and Caki-1-negative control cell lines, followed by transfection and G418 screening of the Caki-1 cells. We measured the expression and enzyme activity of G6PD in the cells by real-time RT-PCR, determined the cell migration phenotypes by Transwell assay, and detected the expressions of p-STAT3 and STAT3 by Western blot.Results Morphologically normal Caki-1-G6PD siRNA and Caki-1-negative control cells were seen under the fluorescence microscope. With GFP expression as a marker, the transfection efficiency rate of the cells was 45－55%. The density of the adherent cells at 48 hours was 90% and their transfection efficiency rate was over 60%. Compared with the Caki-1-negative control cells, the Caki-1-G6PD siRNA cells showed significant decreases in the expressions of Caki-1-G6PD mRNA and protein (P<0.01), enzyme activity (P<0.05), relative count of migratory cells (64.0±4.2 vs 30.0±2.9, P<0.01), and the ratio of p-STAT3/STAT3 (0.45±0.05 vs 0.24±0.01, P<0.01).Conclusion The Caki-1-G6PD siRNA cell line with stable G6PD knockdown and a lower migration ability was first successfully constructed, and the decreased migration ability induced by G6PD knockdown is associated with the STAT3 signal, which is contributive to an insight into the functional and molecular mechanisms of G6PD in the development and progression of ccRCC as well as to finding intervention targets for the treatment of ccRCC.

Influence of 3,3'-diindolylmethane on expression of inflammatory cytokines in periodontal ligament cells induced by lipopolysaccharide / 上海交通大学学报（医学版）

Objective·To investigate the effect of 3,3'-diindolylmethane (DIM) on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLCs) induced by lipopolysaccharide (LPS) and to study the related mechanism.Methyls· hPDLCs were isolated and cultured,and CCK-8 method was used to detect the effect of DIM on the proliferation of hPDLCs.hPDLCs were randomly divided into 4 groups:blank group (without LPS and DIM),LPS group (10 μg/mL LPS),10 μg/mL LPS+6.25 μg/mL DIM,10 μg/mL LPS+12.50 μg/mL DIM.The cells of all groups were cultured for 12 h.The protein levels of TNF-α,IL-1β and IL-6 in supernatant were detected by enzyme linked immunosorbent assay.The change of mitogenactivated protein kinase (MAPK) and nuclear factor κB (NF-κB) signaling pathways were detected by Western blotting.Results· The cell viability was not affected when the DIM concentration was less than 50 μmol/L (P＞0.05).DIM at 6.25 and 12.50 μg/mL reduced the LPS-induced expression of TNF-α,IL-1β and IL-6 at protein levels (P＜0.05).DIM inhibited the activation of the NF-κB signaling pathway.Conclusion· DIM can reduce the LPS-induced inflammatory cytokine expression in hPDLCs via restraining the activation of the NF-κB signaling pathway.

An atypical case of mitochondrial acetoacetyl-CoA thiolase deficiency

Methylacetoacetyl-CoA thiolase deficiency (T2 deficiency) is a rare congenital and metabolic disease affecting the ketone body and isoleucine metabolism. The typical symptoms are refractory metabolic acidosis, in which large amounts of 2-methyl-3-hydroxybutyry1 carnitine, 2-methyl-3-hydroxybutyrate and tiglylglycine are often detected in the blood and urine. We herein describe an atypical case of T2 deficiency with a high level of 3-hydroxybutyrate and a low level of 2-methyl-3-hydroxybutyrate in the urine. Such a case was diagnosed by urinary organic analysis in combination with gene mutation evaluation. Organic acids in the urine were measured using a gas chromatography mass spectrometer and all exons were sequenced via deep sequencing. Molecular biology analysis confirmed the presence of a homozygous mutation in the acetyl-CoA acetyltransferase 1 (ACAT1) gene. The patient received a special diet of deeply hydrolyzed protein milk powder and raw corn starch. She was followed about 6 months. There were no ketoacidotic episodes and hypoglycemia even when she had fever. In conclusion, patients with atypical features of T2 deficiency should also be investigated early. Gas chromatography mass spectrometry and next-generation full exome sequencing may be helpful in diagnosis.

Research Progress of Novel Small Molecule Drugs in the Treatment of Chronic Lymphocytic Leukemia -Review / 中国实验血液学杂志

Chronic lymphocytic leukemia (CLL), the most frequent adult leukemia in Western population, is characterized by accumulation of mature-looking CD5/19/23B cells in peripheral blood, bone marrow, and lymphatic organs. Over the last 20 years, there has been a dramatic change in therapy for CLL, the complete response rate increased from the initial <5% to the current 40%-50%, this remarkable improvement has been attributable to combination of chemoimmunotherapy agents that have contributed to the backbone of therapy for patients with CLL. Especially over the past 5 years, there has been an explosion of new active agents that provide a very effective solution for patients with recurrent/refractory disease as well as those who harbor poor cytogenetic abnormalities. This review focuses on some of the novel small molecule drugs that have either been approved or are at the forefront of clinical development in the treatment of patients with CLL, including tyrosine kinase inhibitior ibrutinib, PI3K inhibitor idelalisib, Syk inhibitor, BCL-2 inhibitor and so on.