Read-through circular RNA rt-circ-HS promotes hypoxia inducible factor 1α expression and renal carcinoma cell proliferation, migration and invasiveness / 北京大学学报(医学版)

OBJECTIVE@#To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.@*RESULTS@#We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.@*CONCLUSION@#The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.

Effects of paternal age on pregnancy outcomes in frozen-thaw embryo transfer cycles / 现代泌尿外科杂志

【Objective】 To observe the effects of paternal age on the pregnancy outcomes in frozen embryo transfer (FET) cycles. 【Methods】 The clinical data of two groups after propensity score matching (PSM) were retrospectively analyzed, including 738 cycles in the 0.05). The clinical pregnancy rate (52.2%vs. 67.2%) and live birth rate (41.1% vs. 57.2%) decreased in the 40-60 year group compared with those in the 0.05). 【Conclusion】 Advanced paternal age decreases clinical pregnancy rate and live birth rate.

Effect of Sodium Salicylate on Calcium Currents and Exocytosis in Cochlear Inner Hair Cells: Implications for Tinnitus Generation / 神经科学通报·英文版

Sodium salicylate is an anti-inflammatory medication with a side-effect of tinnitus. Here, we used mouse cochlear cultures to explore the effects of salicylate treatment on cochlear inner hair cells (IHCs). We found that IHCs showed significant damage after exposure to a high concentration of salicylate. Whole-cell patch clamp recordings showed that 1-5 mmol/L salicylate did not affect the exocytosis of IHCs, indicating that IHCs are not involved in tinnitus generation by enhancing their neuronal input. Instead, salicylate induced a larger peak amplitude, a more negative half-activation voltage, and a steeper slope factor of Ca2+ current. Using noise analysis of Ca2+ tail currents and qRT-PCR, we further found that salicylate increased the number of Ca2+ channels along with CaV1.3 expression. All these changes could act synergistically to enhance the Ca2+ influx into IHCs. Inhibition of intracellular Ca2+ overload significantly attenuated IHC death after 10 mmol/L salicylate treatment. These results implicate a cellular mechanism for tinnitus generation in the peripheral auditory system.

Effect of Buyi Pishen acupuncture on inflammatory factor and cartilage matrix in adjuvant arthritis rats / 中国针灸

OBJECTIVE@#To observe the effect of Buyi Pishen acupuncture (acupuncture for invigorating spleen and kidney) on inflammatory factor and synovial cartilage matrix in adjuvant arthritis (AA) rats, and to explore the mechanism of acupuncture for rheumatoid arthritis (RA).@*METHODS@#A total of 60 clean male Wistar rats were randomized into a normal group, a model group, a tripterygium wilfordii polyglycoside tablet (TWP) group and an acupuncture group, 15 rats in each group. Rats in the model group, the TWP group and the acupuncture group received intradermal injection of Freund's complete adjuvant (FCA) at right hind foot pad to induce the AA model. TWP suspension of 8 mg/kg was given by gavage in the TWP group. Acupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and right "Housanli" (ST 36), "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34) in the acupuncture group, 15 min a time, once a day. The intervention was given 15 days in both TWP group and acupuncture group. The foot-pad swelling degree before modeling, before and after intervention and the arthritis index (AI) score before and after intervention were calculated; the serum levels of interleukin (IL)-1β, IL-4, IL-10 and tumor necrosis factor-α (TNF-α) were detected by ELISA method; the ultrastructure and histomorphological changes of synovium issue were observed by transmission electron microscope and HE staining; the positive expression of matrix metalloproteinase (MMP)-3 and MMP-9 in synovium issue was detected by immunohistochemistry method.@*RESULTS@#Before intervention, foot-pad swelling degree of the model group, the TWP group and the acupuncture group was increased compared with the normal group (P<0.01). After intervention, foot-pad swelling degree and AI score were increased compared with the normal group (P<0.01), foot-pad swelling degree and AI scores in the TWP group and the acupuncture group were lower than the model group (P<0.05), and those in the acupuncture group were decreased compared with the TWP group (P<0.05). The model group exhibited unclear nuclear membrane of synovial cells, chromatin pyknosis, massive inflammatory cell infiltration and hyperplasia in synovial tissue; the TWP group and the acupuncture group exhibited clear and smooth nuclear membrane of synovial cells, inapparent chromatin pyknosis, less inflammatory cell infiltration and hyperplasia in synovial tissue, the acupuncture group exhibited less matrix destruction as well. Compared with the normal group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were increased (P<0.01), while serum levels of IL-4 and IL-10 were decreased (P<0.01) in the model group. Compared with the model group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05, P<0.01), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the TWP group and the acupuncture group; compared with the TWP group, serum level of TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the acupuncture group.@*CONCLUSION@#Buyi Pishen acupuncture can effectively improve the injury of articular cartilage in AA rats, its mechanism maybe related to reducing the inflammatory reaction in synovium and inhibiting the degradation of articular cartilage matrix.

Efficacy of Lauromacrogol Injection for Ablation of Benign Predominantly Cystic Thyroid Nodules and Related Factors: A Prospective Study

Objective@#To prospectively evaluate the efficacy of lauromacrogol injection for ablation (LIA) of benign predominantly cystic thyroid nodules and its related factors. @*Materials and Methods@#A total of 142 benign predominantly cystic thyroid nodules (median volume, 12.5 mL; range, 0.4– 156 mL) in 137 patients (male:female sex ratio, 36:101; mean age ± standard deviation [SD], 49 ± 13 years) were treated with LIA after being confirmed as benign via cytology. The volume reduction rate (VRR) of the nodules and cosmetic score were evaluated during follow-up at 1, 3, and 6 months after treatment and every 6 months thereafter. A VRR of ≥ 50% at the 12-month follow-up was considered to indicate effective treatment. The associations between the clinical factors and nodular ultrasound features, including the initial nodule volume, proportion of solid components, vascularity grade and ineffective treatment (VRR of < 50% at the 12-month follow-up), and regrowth were analyzed. @*Results@#All patients completed follow-up for at least 12 months. The average ± SD follow-up period was 32 ± 11 months (range, 12–54 months). The effective treatment rate was 73.2% (104/142), while the regrowth rate was 12.0% (17/142) at the last follow-up. Grade 2–3 intranodular vascularity in the solid components of the nodules was the only independent factor associated with ineffective treatment, with an odds ratio (reference category, grade 0–1) of 3.054 (95% confidence interval, 1.148–8.127) (p = 0.025). @*Conclusion@#LIA is an effective treatment for predominantly cystic thyroid nodules. Grade 2–3 intranodular vascularity in the solid components of nodules is the only independent risk factor for ineffective LIA.

The application of organoid models in diabetes research / 中华内分泌代谢杂志

Organoids are complex tiny organ-like model systems formed by three-dimensional culture in vitro, based on the self-renew and self-organization of stem cells. This article reviewed the recent progress in organoids construction from tissues involved in the regulation of glucose homeostasis and chronic diabetic microvascular complications, and their applications in diabetes mellitus. Organoid technology is expected to further promote the progress of diabetes research in disease modeling, personalized medicine, and regenerative medicine.

EGFR tyrosine kinase inhibitor HS-10296 induces autophagy and apoptosis in triplenegative breast cancer MDA-MB-231 cells / 浙江大学学报·医学版

OBJECTIVE@#To investigate the inhibitory effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI) HS-10296 on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells and explore the possible molecular mechanism.@*METHODS@#MDA-MB-231 cells were treated with HS-10296 for 24, 48, or 72 h, and CCK-8 assay was used to assess the changes in the cell viability. The inhibitory effect of HS-10296 on cell proliferation was determined by clonogenic assay. JC-1 and flow cytometry were employed for analyzing the cell apoptosis, and the ultrastructure of the cells was observed under electron microscope. After pretreatment with autophagy inhibitor chloroquine (CQ), MDA-MB-231 cells were divided into control group, CQ treatment group, HS-10296 (4 and 6 μmol/L) treatment groups and combined treatment groups, and the sensitivity of the treated cells to HS-10296 was determined using CCK-8 assay. The effects of HS-10296 on EGFR pathway and apoptosis- and autophagy-related proteins in MDA-MB-231 cells were investigated using Western blotting.@*RESULTS@#HS-10296 significantly inhibited the proliferation of MDA-MB-231 cells with IC values at 24, 48 and 72 h of 8.393, 2.777 and 2.016 μmol/L, respectively. JC-1 and flow cytometry showed that HS-10296 induced obvious apoptosis of MDA-MB-231 cells, which showed an apoptosis rate of (21.63 ± 2.97)% following treatment with 8 μmol/L HS-10296. Autophagy vesicles were observed in the cells treated with HS-10296 under electron microscope. In MDA-MB-231 cells pretreated with CQ, inhibition of autophagy significantly enhanced HS-10296-induced cell death. Western blotting showed that the apoptosis-related protein caspase-3 was activated after HS-10296 treatment to cut its substrate PARP. The expression of autophagy-related protein light chain 3B (LC3B) was significantly enhanced after HS-10296 treatment ( < 0.01), which also resulted in inhibited phosphorylation of EGFR and AKT proteins in the cells.@*CONCLUSIONS@#HS-10296 can inhibit the proliferation and induce autophagy and apoptosis of MDA-MB-231 cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

Effect of Huangqin Qingre Chubi Capsules containing serum on oxidative stress and protein expression of AMPK and FoxO3a in rheumatoid arthritis patients / 中国中药杂志

To study the effect of Huangqin Qingre Chubi Capsules containing serum on the protein expressions of AMPK and FoxO3 a in peripheral blood mononuclear cells of patients with rheumatoid arthritis(RA), in order to explore the mechanism of anti-oxidation. Peripheral anticoagulant was collected from patients and normal people. Monocytes(PBMC) were isolated through density gradient centrifugation, and the logarithmic phase cells were cultured. Drug containing serum was prepared through intragastric admini-stration to SD rats. The rats were divided into five groups, namely normal group, model group, AMPK blocker group(compound C 10 μmol·L~(-1)), medium-dose HQC+AMPK blocker group, and middle-dose HQC group. The cell inhibition rate was calculated by MTT method. The levels of IL-1β, IL-4, LPO, MDA, SOD and TAOC were detected by ELISA. The expressions of AMPK, p-AMPK, p-FoxO3 a and FoxO3 a were detected by Western blot. The HQC containing serum had an inhibitory effect on human monocytes in peripheral blood. The best concentration was observed in middle-dose HQC, and the best time was 24 hours. Middle-dose HQC group was better than model group, AMPK blocker group and middle-dose HQC + AMPK blocker group in terms of increase of SOD, p-AMPK, p-FoxO3 a and decrease of LPO. It was better than model group and AMPK blocker group in terms of increase of IL-4, TAOC, AMPK, FoxO3 a and decrease of IL-1β, MDA. The differences were statistically significant(P<0.05 or P<0.01). The HQC containing serum may increase the levels of TAOC and SOD, decrease the level of MDA and LPO, activate AMPK, directly phosphorylate FOXO3 a, enhance its transcriptional activity, and improve the state of oxidative stress in RA patients.

Adefovir Dipivoxil plus Chinese Medicine in HBeAg-Positive Chronic Hepatitis B Patients: A Randomized Controlled 48-Week Trial / 中国结合医学杂志

OBJECTIVE@#To evaluate the effects of a 48-week course of adefovir dipivoxil (ADV) plus Chinese medicine (CM) therapy, namely Tiaogan Jianpi Hexue () and Tiaogan Jiedu Huashi () fomulae, in hepatitis B e antigen (HBeAg)-positive Chinese patients.@*METHODS@#A total of 605 HBeAg-positive Chinese CHB patients were screened and 590 eligible participants were randomly assigned to 2 groups in 1:1 ratio including experimental group (EG, received ADV plus CM) and control group (CG, received ADV plus CM-placebo) for 48 weeks. The major study outcomes were the rates of HBeAg and HBV-DNA loss on week 12, 24, 36, 48, respectively. Secondary endpoints including liver functions (enzymes and bilirubin readings) were evaluated every 4 weeks at the beginning of week 24, 36, and 48. Routine blood, urine, and stool analyses in addition to electrocardiogram and abdominal B scan were monitored as safety evaluations. Adverse events (AEs) were documented.@*RESULTS@#The combination therapy demonstrated superior HBeAg loss at 48 weeks, without additional AEs. The full analysis population was 560 and 280 in each group. In the EG, population achieved HBeAg loss on week 12, 24, 36, and 48 were 25 (8.90%), 34 (12.14%), 52 (18.57%), and 83 (29.64%), respectively; the equivalent numbers in the CG were 20 (7.14%), 41 (14.64%), 54 (19.29%), and 50 (17.86%), respectively. There was a statistically significant difference between these group values on week 48 (P<0.01). No additional AEs were found in EG. Subgroup analysis suggested different outcomes among treatment patterns.@*CONCLUSION@#Combination of CM and ADV therapy demonstrated superior HBeAg clearance compared with ADV monotherapy. The finding indicates that this combination therapy may provide an improved therapeutic effect and safety profile (ChiCTR-TRC-11001263).

EGFR tyrosine kinase inhibitor HS-10296 induces autophagy and apoptosis in triplenegative breast cancer MDA-MB-231 cells / 南方医科大学学报

OBJECTIVE@#To investigate the inhibitory effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFRTKI) HS-10296 on the proliferation of triple-negative breast cancer (TNBC) MDA-MB-231 cells and explore the possible molecular mechanism.@*METHODS@#MDA-MB-231 cells were treated with HS-10296 for 24, 48, or 72 h, and CCK-8 assay was used to assess the changes in the cell viability. The inhibitory effect of HS-10296 on cell proliferation was determined by clonogenic assay. JC-1 and flow cytometry were employed for analyzing the cell apoptosis, and the ultrastructure of the cells was observed under electron microscope. After pretreatment with autophagy inhibitor chloroquine (CQ), MDA-MB-231 cells were divided into control group, CQ treatment group, HS-10296 (4 and 6 μmol/L) treatment groups and combined treatment groups, and the sensitivity of the treated cells to HS-10296 was determined using CCK-8 assay. The effects of HS-10296 on EGFR pathway and apoptosis- and autophagy-related proteins in MDA-MB-231 cells were investigated using Western blotting.@*RESULTS@#HS-10296 significantly inhibited the proliferation of MDA-MB-231 cells with IC values at 24, 48 and 72 h of 8.393, 2.777 and 2.016 μmol/L, respectively. JC-1 and flow cytometry showed that HS-10296 induced obvious apoptosis of MDA-MB-231 cells, which showed an apoptosis rate of (21.63 ± 2.97)% following treatment with 8 μmol/L HS-10296. Autophagy vesicles were observed in the cells treated with HS-10296 under electron microscope. In MDA-MB-231 cells pretreated with CQ, inhibition of autophagy significantly enhanced HS-10296-induced cell death. Western blotting showed that the apoptosis-related protein caspase-3 was activated after HS-10296 treatment to cut its substrate PARP. The expression of autophagy-related protein light chain 3B (LC3B) was significantly enhanced after HS-10296 treatment ( < 0.01), which also resulted in inhibited phosphorylation of EGFR and AKT proteins in the cells.@*CONCLUSIONS@#HS-10296 can inhibit the proliferation and induce autophagy and apoptosis of MDA-MB-231 cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

Gefitinib inhibits glycolysis and induces programmed cell death in non-small cell lung cancer cells / 南方医科大学学报

OBJECTIVE@#To observe the cell death pattern induced by gefitinib in non-small cell lung cancer A549 and H1975 cells and explore the possible mechanism in light of glycolysis.@*METHODS@#The inhibitory effects of gefitinib at 20, 30, or 40 μmol/L in A549 cells and at 20, 40, or 80 μmol/L in H1975 cells were examined using MTT assay. The changes of lactic acid level in the cells were determined with a lactic acid kit, and the expression levels of glycolysis-related proteins (PKM2 and HK2) and the proteins in PI3K-Akt-mTOR signaling pathway were detected using Western blotting. 2-NBDG was used for detecting glucose uptake capacity of the cells, and ATP kit was used to detect the intracellular ATP level. The mitochondrial membrane potential of the cells was examined with the JC-1 kit, and cell apoptosis was analyzed with Annexin V-FITC/PI double staining. The relative expression levels of the apoptotic proteins Bax and Bcl-2 and the autophagy marker protein LC3B were detected with Western blotting.@*RESULTS@#MTT assay showed that gefitinib inhibited the proliferation of A549 and H1975 cells in a time- and dose-dependent manner ( < 0.05). The IC of gefitinib at 24, 48 and 72 h was 48.6, 28.6 and 19.7 μmol/L in A549 cells and was 321.6, 49.1 and 14.6 μmol/L in H1975 cells, respectively. Gefitinib significantly lowered intracellular lactic acid level of the cells ( < 0.05) and down-regulated the expressions of PKM2 and HK2 proteins ( < 0.05) and PI3K-Akt-mTOR signaling pathway-associated proteins ( < 0.05). Gefitinib obviously inhibited glucose uptake and ATP levels in both A549 and H1975 cells ( < 0.05). Treatment with gefitinib induced obviously enhanced apoptosis in the cells, resulting in apoptosis rates of (10.77± 1.0)%, (14.5±0.4)%, (17.4±0.2)% and (32.1±0.6)% at 0, 20, 30 and 40 μmol/L in A549 cells ( < 0.05) and of (10.5±0.6)%, (13.2± 0.92)%, (18.9±0.98)% and (35.1±1.4)% at 0, 20, 40 and 80 μmol/L in H1975 cells, respectively ( < 0.05). The protein expression of Bax increased and that of Bcl-2 decreased following gefitinib treatment in the cells ( < 0.05). Gefitinib significantly increased autophagy in A549 and H1975 cells as shown by increased LC3B expressions following the treatment ( < 0.05).@*CONCLUSIONS@#Gefitinib can inhibit the proliferation, induce apoptosis and increase autophagy in A549 and H1975 cells. Gefitinib induces apoptosis of the cells possibly by affecting glycolysis and PI3K-Akt-mTOR signaling pathway.

Treatment of Pilon fractures complicated with soft tissue injury by plate and lag screw fixation via posterolateral approach / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the clinical effects of plate and lag screw fixation for treatment of Pilon fractures complicated with soft tissue injury via posterolateral approach.</p><p><b>METHODS</b>From May 2013 to June 2016, 25 patients with Pilon fractures complicated with soft tissue injury underwent open reduction and internal fixation via posterolateral approach. There were 15 males and 10 females, aged from 25 to 61 years old with an average of(39.6±0.2) years. Plate and lag screw fixation were used in operation. Healing of soft tissue contusion and abrasions in the ankle wounds and injuries were observed after operation. The Burwell-Charnley standard was applied to assess the quality of fracture reduction and the AOFAS Ankle foot scoring system(total score 100 points) was used to evaluate the clinical effects.</p><p><b>RESULTS</b>All the patients were followed up from 6 to 24 months with an average of 12 months. All operative wounds and soft tissue injuries were healed. According to the Burwell-Charnley standard, 22 cases obtained excellent results with anatomic reduction, while 2 cases were dissatisfied, and 1 case poor. The AOFAS ankle foot scores were 90.2±7.5 on average, with 20 cases of excellent results, 3 good, 2 fair.</p><p><b>CONCLUSIONS</b>Plate and lag screw fixation by posterolateral approach in treating Pilon fracture complicated with soft tissue injury shows advantage of avoiding injury to the anteromedial skin and soft tissue, provides forceful fixation without further injury.</p>

Preliminary study on the clinical application of temporary treatment denture in difficult edentulous cases before complete denture restoration / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the clinical effects of treatment denture on difficult edentulous cases before complete denture restoration.</p><p><b>METHODS</b>Thirty-six patients who experienced unsuccessful restoration of conventional complete dentures were included in this study. Treatment dentures were fabricated to solve issues such as abnormal occlusion, tissue surface problems, and neuromuscular dysfunction of the stomatognathic system caused by systemic diseases. The final complete dentures were fabricated by duplicating the treatment dentures. Jaw relation index, stability, and retention were evaluated at different stages. Oral health-related quality of life was measured using the Chinese version of Oral Health Impact Profile for edentulous subjects (OHIP-EDENT).</p><p><b>RESULTS</b>Among the 36 patients, 33 successfully completed the final restoration with positive effects.</p><p><b>CONCLUSIONS</b>Treatment denture is an effective pre-restorative option that can be used to correct abnormal occlusion, improve tissue surface problems, and aid in neuromuscular rehabilitation training. Treatment dentures contribute to the successful restoration of the final complete dentures and is worthy of clinical applications.</p>

Knocking-out of HIF1α gene by CRISPR/cas9 inhibits proliferation and invasiveness of prostate cancer DU145 cells / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the role of HIF1α gene in prostate cancer cell line DU145 by knocking it out with a novel gene-editing tool CRISPR/cas9 system.</p><p><b>METHODS</b>A CRISPR/cas9 system with two sgRNAs targeting exon 1 of the HIF1α gene was constructed for the knock out experiment. CCK8 assay and transwell experiment were carried out to assess the effect of the knock out on the proliferation, migration and invasiveness of DU145 cells.</p><p><b>RESULTS</b>The efficiency of gene-targeting was measured through a T7E1 assaying and sequence analysis, which confirmed that the partial knock out was successful and has led to a significant decrease in the expression of HIF1α and inhibition of cell proliferation, migration and invasiveness.</p><p><b>CONCLUSION</b>A CRISPR/cas9 system for the knock out of HIF1α has been successfully constructed, which could inhibit the proliferation and migration of DU145 cells. The system can facilitate further studies of the HIF1α gene and its roles in tumorigenesis.</p>

Chinese Herbal Medicine Combined with Entecavir for HBeAg Positive Chronic Hepatitis B: Study Protocol for a Multi-Center, Double-Blind Randomized-Controlled Trial / 中国结合医学杂志

<p><b>BACKGROUND</b>The domestic prevalence of chronic hepatitis B (CHB) in China is 7.18% in 2006, imposing great societal healthcare burdens. Nucleot(s)ide analogues (NUCs) anti-hepatitis B virus (HBV) therapies are widely applied despite the relatively low rate of seroconversion and high risk of drug-resistant mutation. More effective treatments for CHB deserve further explorations. Combined therapy of NUCs plus Chinese herbal medicine (CHM) is widely accepted in China, which is recognized as a prospective alternative approach. The study was primarily designed to confirm the hypothesis that Tiaogan-Yipi Granule (, TGYP) or Tiaogan-Jianpi-Jiedu Granule (, TGJPJD) plus entecavir tablet (ETV) was superior over ETV monotherapy in enhancing HBeAg loss rate.</p><p><b>METHODS</b>The study was a nationwide, large-scale, multi-center, double-blind, randomized, placebo-controlled trial with a designed duration of 108 weeks. A total of 16 hospitals and 596 eligible Chinese HBeAg positive CHB patients were enrolled from November 2012 to September 2013 and randomly allocated into 2 groups in 1:1 ratio via central randomization system: experimental group (EG) and control group (CG). Subjects in EG received CM formulae (TGYP or TGJPJD, 50 g per dose, twice daily) plus ETV tablet (or ETV placebo) 0.5 mg per day in the first 24 weeks (stage 1), and CHM granule plus ETV tablet (0.5 mg per day) from week 25 to 108 (stage 2). Subjects in CG received CHM Granule placebo plus ETV tablet (0.5 mg per day) for 108 weeks throughout the trial. The assessments of primary outcomes (HBV serum markers and HBV-DNA) were conducted by a third-party College of American Pathologists (CAP) qualified laboratory. Adverse effects were observed in the hospitals of recruitment.</p><p><b>DISCUSSION</b>The study was designed to compare the curative effect of CM plus ETV and ETV monotherapy in respect of HBeAg loss, which is recognized by the European Association for the Study of the Liver as "a valuable endpoint". We believe this trial could provide a reliable status for patients' "journey" towards durable responses after treatment discontinuation. The trial was registered before recruitment on Chinese Clinical trial registry (No. ChiCTR-TRC-12002784, Version 1.0, 2015/12/23).</p>

Prognostic significance of combined TERT and IDH gene mutation analysis in diffusely infiltrating gliomas / 中华病理学杂志

Objective@#To investigate the status and prognostic significance of TERT and IDH1/2 genes mutations in diffusely infiltrating gliomas.@*Methods@#Hot spot mutations of TERT and IDH1/2 genes were detected by DNA sequencing in 236 cases of gliomas at West China Hospital from 2012 to 2016, including pilocytic astrocytoma (WHO grade Ⅰ, 16 cases), diffuse astrocytoma and oligodendroglioma (WHO grade Ⅱ, 89 cases), anaplastic astrocytoma and oligodendroglioma (WHO grade Ⅲ, 72 cases) and glioblastoma (WHO grade Ⅳ, 59 cases). The prognostic significance of TERT and IDH1/2 hot spot mutations was evaluated.@*Results@#No IDH or TERT mutations were detected in pilocytic gliomas. TERT promoter mutation frequency was higher in patients aged ≥40 years(60.8%, 93/153) than in patients aged <40 years (32.8%, 22/67; P<0.01). TERT promoter mutation rate was also significantly higher in oligodendroglioma (87.5% , 56/64) than that in astrocytoma(37.8%, 59/156; P<0.01). Young age (<40 years), oligodendroglioma and IDH1 mutation were favorable prognostic factors for diffusely infiltrating astrocytic and oligodendroglial tumors. TERT mutation alone was not of prognostic significance. Diffusely infiltrating astrocytic and oligodendroglial tumors were divided into four molecular subtypes according to TERT and IDH1 mutation status: IDH(+ )/TERT(+ ), IDH(+ )/TERT(-), IDH(-)/TERT(-) and IDH(-)/TERT(+ ). There was significant prognostic difference among the 4 subtypes.@*Conclusions@#Combined IDH and TERT gene mutation analysis may be useful for prognostic subgrouping. Notably, IDH1 wild-type cases can be further subdivided into TERT(+ ) or (-) subgroups with significant prognostic difference.

Evaluation of effect of Hibiscus paste external application combined with traditional Chinese medicine on inflammatory indices of patients with active rheumatoid arthritis based on random walk model / 中国免疫学杂志

Objective:To evaluate the effect of Hibiscus paste external application combined with traditional Chinese medicine on the inflammatory markers in patients with active rheumatoid arthritis (RA). Methods: The patient information was collected by the data processing system of hospitalized medical records in the First Affiliated Hospital of Anhui University of traditional Chinese medicine. The medical records of patients with rheumatoid arthritis in the Department of Rheumatology from May 2012 to December 2016 were collected. Patients were divided into experimental group and control group according to whether Furong ointment was applied. The random walk model was used to evaluate the influence of Furong ointment combined with traditional Chinese medicine on inflammatory index,erythrocyte sedimentation rate ( ESR) and high sensitivity C reactive protein ( Hs-CRP) in patients with active rheumatoid arthritis. Results: A total of 4 832 patients with rheumatoid arthritis were in accordance with the requirements of the study, including 2 579 in the experimental group and 2 253 in the control group. Baseline data analysis showed that there was no significant difference in general condition, frequency of oral Chinese medicine and core prescriptions between two groups ( P＞0. 05 ) . In the experimental group,the maximum ESR of inflammation index was 968,the walking rate was 3 756,the positive growth rate of walking was 0. 257 7,the ratio was 3. 88,the random fluctuation power law value was 0. 364 2±0. 124 6,the comprehensive evaluation index in-creased by 0. 479 9,and the comprehensive evaluation index was 2 017 times. In the control group,the maximum ESR of inflammation index was 398,the number of walking steps was 2 251,the positive growth rate of walking was 0. 176 8,the ratio was 5. 66,the random fluctuation power law value was 0. 173 5±0. 128 8,the comprehensive evaluation index increased by 0. 357,and the comprehensive evaluation index was 1 115 times. In the experimental group,the maximum Hs-CRP of inflammation index was 1 523,the walking rate was 5 149,the positive growth rate of walking was 0. 295 8,the ratio was 3. 38,the random fluctuation power law value was 0. 389 5± 0. 108 1,the comprehensive evaluation index increased by 0. 537 4,and the comprehensive evaluation index was 2 834 times. In the control group,the maximum Hs-CRP of inflammation index was 809,the number of walking steps was 3 463,the positive growth rate of walking was 0. 233 6,the ratio was 4. 28,the random fluctuation power law value was 0. 362 9±0. 073 8,the comprehensive evaluation index increased by 0. 448 2,and the comprehensive evaluation index was 1 805 times. Conclusion: Our hospital rheumatology on the treatment of rheumatoid arthritis from the spleen, oral Chinese medicine prescription to Jianpihuashi drugs, Huoxue Tongluo drugs, Qufengchushi drugs,drug detoxification,two groups of patients with comprehensive evaluation index and acceptable interventions are long-range correlation,a better effect than that of inflammatory markers associated with traditional Chinese medicine in patients with Furong ointment the simple use of Chinese herbs.

Myricetin blocks the activation,proliferation and secretion of cardiac fibroblasts induced by TGF-β / 医学研究生学报

Objective It is rarely reported whether myricetin inhibits the activation and function of cardiac fibroblasts and thereby prevents myocardial fibrosis. This study was to investigate the effects of myricetin on the activation,proliferation and secretion of cardiac fibroblasts and its possible molecular mechanisms. Methods Fibroblasts isolated from 1-3 days old rats were cultured and their activation,proliferation and secretion were induced with the transforming growth factor (TGF). The fibroblasts were incubated with myricetin at different concentrations of 1,3,10,30 and 100 μmol/L for 24 hours followed by detection of their proliferation with the CKK8 kit,the transcription levels of fibrotic factors by RT-PCR and the expression levels of α-SMA and signal proteins by immunoflu-orescence staining and Western blot,respectively. Results The expression of α-SMA was significantly up-regulated in the cardiac fi- broblasts of the rats in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups as compared with that in the control group (P<0.05) but down-regulated in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups in com-parison with that in the TGF-β group (P<0.05). At 48 hours,the transcription levels of collagenⅠ,collagenⅢ,fibronectin and con-nective tissue growth factor were markedly higher in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the control group (P<0.05) but lower in the 30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups than in the TGF-β group (P<0.05). The phosphorylation levels of smad2 and smad3 were remarkably elevated in the TGF-β,30 μmol/L myricetin+TGF-β and 100 μmol/L myricetin+TGF-β groups and the expression of smad4 reduced in the TGF-β group as com-pared with the control group (P<0.05). The levels of smad2,smad3 and smad4 were all significantly decreased in the 30 μmol/L myr-icetin+TGF-β and the 100 μmol/L myricetin+TGF-β groups in comparison with the TGF-β group (P<0.05). Conclusion Myricetin suppresses the activation,proliferation and secretion of cardiac fibroblasts induced by TGF-β via inhibiting the smad signaling pathway.

Effect of MCUR1 on proliferation,cell cycle and apoptosis of K562 cells / 军事医学

Objective To investigate the effect of mitochondrial calcium uniporter(MCU)regulator 1(MCUR1)on proliferation,cell cycle and apoptosis of K562 cells and the possible molecular mechanism.Methods Recombinant plasmid vectors containing short hairpin RNAs(shRNAs)targeting MCUR1 were transfected into K562 cells,before the K562 cells stably expressing low MCUR1 were selected with G418.The expression of MCUR1 mRNA was detected by quantitative real-time polymerase chain reaction(qRT-PCR)assays.Western blotting(WB)assays were used to detect the expressions of MCUR1,P53,BAX and BCL2.The proliferation,cell cycle and apoptosis of K562 cells were detected by cell counting kit-8(CCK-8)assays and flow cytometry, respectively.Results The results of qRT-PCR and WB assays revealed that MCUR1 was stably down-regulated at mRNA and protein levels in the K562 cells transfected with shRNAs targeting MCUR1.Knockdown of MCUR1 significantly inhibited the cell proliferation, induced the cell apoptosis, but did not influence the cells cycle.Meanwhile, knockdown of MCUR1 increased the expression of P53 protein and the ratio of protein BAX/BCL2 in K562 cells.Conclusion MCUR1 promotes cell proliferation and inhibits cell apoptosis in K 562 cells.

Expression of CD44, TRIM24, TAGLN-2, ER and PR in breast invasive ductal carcinoma and their clinicopathologic significance / 临床与实验病理学杂志

Purpose To study the expression of biomolecules of PI3K-AKT signaling pathway including CD44,TRIM24,TAGLN-2,ER and PR in breast invasive ductal carcinoma,and to explore their clinicopathologic significance.Methods The expression of CD44,TRIM24,TAGLN-2,ER and PR in 73 cases of breast invasive ductal carcinoma were detected by immunohistochemist~ (IHC) technology.And the relationship between the expression of these biomarkers and the age of patients,tumor size,histological grade,lymph node metastasis was analyzed.Besides,the influence of these biomarkers on prognosis and the relationship between the expression of these biomarkers were also analyzed.Results There was no significant relationship between the expression of these biomarkers and the age of patients,tumor size,histological grade,lymph node metastasis.CD44 expression was positively conelated with TAGLN-2 expression (r =0.311,P =0.007),TRIM24 expression was positively correlated with TAGLN-2 expression (r =0.421,P =0.000).CD44 expression was negatively correlated with ER expression (r =-0.285,P =0.015).ER expression was positively correlated with PR expression (r =0.598,P =0.000).The postoperative 5-year cumulative survival-rate of CD44 positive group was lower than those of CD44 negative group (P =0.002),in contrast,the postoperative 5-year survival rate of ER positive group was higher than those of ER negative group (P =0.026).Conclusion Through PI3K-AKT signaling pathway,CD44 and TRIM24 may up-regulate TAGLN-2 expression,however,CD44 may down-regulate ER and PR expression.CD44 and ER may act an useful predictor for the prognosis of breast invasive ductal carcinoma.Combined detection of CD44,ER and PR may provide additional therapeutic information which may be useful in breast cancer treatment.