Preparation and in vitro evaluation of prostate cancer exosomes containing melittin / 中国药理学通报

Aim To prepare prostate cancer exosomes containing melittin and observe their uptake by prostate cancer cells. Methods Cells treated with starvation for different time were screened for exosome extraction. Exosomes from PC-3 cells were extracted by ultracentrifugation, and the extracted particles were examined by transmission electron microscopy, nanoparticle tracking analyzer(NTA), and Western blot. Melittin exosome system was prepared by repeated freeze-thaw method, incubation at room temperature as well as electroporation, and the size of encapsulation efficiency was measured by centrifugation. A high-performance liquid chromatography(HPLC)method was applied to assay the content of melittin exosomes(exo-mel). Fluorescence inverted microscopy was employed to evaluate the uptake of melittin exosomes by PC-3 cells, DU145 cells as well as LNCaP cells. Results The results of starvation treatment showed that 24 h starvation treatment was the optimal time point. TEM results showed that the exosomes were round or oval in shape with a distinct membranous structure, and the diameter was around 100 nm. The reagent protein concentration for NTA analysis of exosomes was 0.222 g·L-1. The results of Western blot for the marker proteins of exosomes showed that Alix and CD63 were positively expressed, which indicated that the exosomes could be obtained by starvation culture of PC-3 cells and ultracentrifugation. The results of entrapment efficiency showed that the entrapment efficiency of electroporation method was 17.51% ± 2.39%, that of repeated freeze-thaw method was 11.46% ± 1.02%, and that of room temperature incubation method was 3.93% ± 2.44%. The encapsulation efficiency of electroporation was the highest with significant difference(P<0.05). The uptake assay showed that PC-3 cells could efficiently take up exo-mel in a time-dependent manner, and DU145 cells and LNCaP cells also could take up exo-mel over time. Conclusions Exosomes can be accessed by starvation treatment and high-speed centrifugation, and the prostate cancer melittin exosome system prepared by electroporation method could be taken up by prostate cancer cells.

The effect of scrotal versus inguinal orchiopexy on the testicular function of children with clinically palpable, inguinal undescended testis: a randomized controlled trial / 亚洲男科学杂志(英文版)

To compare the impact of the scrotal vs inguinal orchidopexy approach on the testicular function of infants with cryptorchidism, a randomized controlled trial was conducted involving boys who were 6-12 months old at surgery and were diagnosed with clinically palpable, inguinal undescended testis. Between June 2021 and December 2021, these boys at Fujian Maternity and Child Health Hospital (Fuzhou, China) and Fujian Children's Hospital (Fuzhou, China) were enrolled. Block randomization with a 1:1 allocation ratio was employed. The primary outcome was testicular function assessed by testicular volume, serum testosterone, anti-Müllerian hormone (AMH), and inhibin B (InhB) levels. Secondary outcomes included operative time, amount of intraoperative bleeding, and postoperative complications. Among 577 screened patients, 100 (17.3%) were considered eligible and enrolled in the study. Of the 100 children who completed the 1-year follow-up, 50 underwent scrotal orchidopexy and 50 underwent inguinal orchidopexy. The testicular volume, serum testosterone, AMH, and InhB levels in both groups increased markedly after surgery (all P < 0.05), but there were no apparent differences between groups at 6 months and 12 months after operation (all P > 0.05). No differences between the scrotal and inguinal groups were noted regarding the operative time ( P = 0.987) and amount of intraoperative bleeding ( P = 0.746). The overall complication rate (2.0%) of the scrotal group was slightly lower than that of the inguinal group (8.0%), although this difference was not statistically significant ( P > 0.05). Both scrotal and inguinal orchiopexy exerted protective effects on testicular function in children with cryptorchidism, with similar operative status and postoperative complications. Scrotal orchiopexy is an effective alternative to inguinal orchiopexy in children with cryptorchidism.

Research progress on the interactions between gut bacterial β-glucuronidases and Chinese herbal medicines / 药学学报

In traditional oral practice, the presystemic interactions with gut microbiota is an important mechanism underlying the holistic health benefits of Chinese herbal medicines (CHMs), making the study of CHMs distinct from the research of Western medicines of which the systemic exposure (level in blood) is the starting point and the core. Gut microbial metabolism complements host metabolism in maintaining metabolic homeostasis of many biologically important endogenous molecules and the disposition of numerous exogenous compounds. Among them, the widely distributed gut bacterial β-glucuronidases (BGUSs) coordinate with host UDP-glucuronosyltransferases (UGTs) to play a role in the occurrence and intervention of diseases by affecting the glucuronidation homeostasis and altering the intestinal local and/or systemic exposure of endogenous compounds and xenobiotics. On one hand, many ingredients of CHMs undergo enterohepatic circulation; On the other hand, CHMs can act on BGUSs directly or indirectly change the distribution and function of BGUSs through reprogramming gut microbiome. The multiple interactions between BGUSs and CHMs may play an important role in the overall therapeutic benefits of CHMs. This work firstly summarizes the latest research progress on BGUSs; then the physiological, pathological and pharmacological significance of BGUSs are exemplified with representative endogenous and exogenous compounds from the aspects of nutrient utilization, metabolic homeostasis, and therapeutic response based on the varied substrate spectra of BGUSs; finally, the scattered data in literature were integrated to summarize the multiple interactions between BGUSs and CHMs, highlighting the important role of BGUSs in the holistic actions of CHMs.

Renoprotective Effect of Sanjiao Qushi Prescription Against Cationic Bovine Serum Albumin Induced Membranous Nephropathy Mice Model and Its Influence on Nrf2/HO-1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the renoprotective effects that Sanjiao Qushi prescription ameliorates cationic bovine serum albumin （C-BSA） induced membranous nephropathy（MN） in mice model and its influence on nuclear factor erythroid-2-related factor-2（Nrf2）/heme oxygenase-1（HO-1） signaling pathway. Method:Sixty female BALB/c mice were randomly divided into the normal group（<italic>n</italic>=10） and the model group（<italic>n</italic>=50）. The mice in the model group received C-BSA injection via tail vein （6.5 mg·kg^-1）. The mice that were successfully modeled were randomized into the model group， the low dose Sanjiao Qushi prescription group（3.71 g·kg^-1）， the high dose Sanjiao Qushi prescription group（7.42 g·kg^-1） and benazepril hydrochloride group（1.3 mg·kg^-1）. And they were administered with the corresponding medicine by gavage once a day for four consecutive weeks. 24 hour-urine protein quantitation were performed before C-BSA injection and after C-BSA injection as well as the medicine gavage. When the treatment was finished， all of the mice were sacrificed and the biochemical indicators such as serum creatinine（SCr）， blood urea nitrogen（BUN）， triglyceride（TG）， total cholesterol（TC）， total protein（TP） and albumin（Alb） were measured. And the renal pathological morphology changes were observed by light microscope with hematoxylineosin（HE）， Masson and periodic acid-silver metheramine（PASM） staining. The deposition of immunoglobulin G（IgG） in the glomerulus was detected by fluorescence microscope. The expression of reactive oxygen species （ROS） of kidney was detected by fluorescence immunoassay. The protein expression levels of Nrf2 in cell nucleus and cytoplasm and the downstream protein factors HO-1 and NADH quinone acceptor oxidoreductase 1（NQO1） were detected by Western blot. Result:Compare to normal group， the levels of 24 hour-urine protein quantitation， TG and TC significantly increased in model group（<italic>P</italic><0.01）， while TP and Alb levels significantly decreased（<italic>P</italic><0.01）. The model group exhibited enlarged volume of glomerular， significantly thickened glomerular basement membrane（GBM）， fuchsinophilic protein deposition and spike formation through light microscope. Immunofluorescence staining for the model group exhibited granular deposition of IgG along the capillary wall. The expression of ROS in kidney significantly increased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly increased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly decreased（<italic>P</italic><0.01）.The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Compared to model group， the levels of 24 hour-urine protein quantitation， TG and TC significantly decreased in each treated group（<italic>P</italic><0.01）， TP and Alb levels significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The pathological damages alleviated obviously. The expression of ROS in kidney significantly decreased（<italic>P</italic><0.01）. The protein expression levels of Nrf2 in cell nucleus significantly decreased（<italic>P</italic><0.01）， while Nrf2 in cytoplasm significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The protein expression levels of HO-1 and NQO1 significantly increased（<italic>P</italic><0.01）. Conclusion:Sanjiao Qushi prescription worked on MN mice possibly by regulating related proteins in the Nrf2/HO-1 signaling pathway and relieving oxidative stress， thus decreasing 24 hour-urine protein and blood lipid， increasing serum protein， and alleviating the pathological damages to protect renal function and delay progress of the disease.

Molecular Mechanism of Platycodonis Radix- Armeniacae Semen Amarum in Treating Lung Carcinoma Based on Network Pharmacology / 中国实验方剂学杂志

Objective:To screen out the active components of Platycodonis Radix-Armeniacae Semen Amarum, predict the targets and signaling pathways, construct the "multi-components, multi-targets and multi-pathways" interaction network and further investigate their molecular mechanism for the treatment of lung carcinoma based on network pharmacology. Method:Active components and corresponding targets of Platycodonis Radix-Armeniacae Semen Amarum were obtained through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and literature consultation. Therapeutic targeted genes of Platycodonis Radix-Armeniacae Semen Amarum in the treatment of lung carcinoma were obtained from UniProt database and Genecards database. The "components-targets" network was constructed by using Cytoscape 3.6.0 software, and the protein-protein interactions network was constructed by String database and "Generate Style From Statistics" tool in Cytoscape software. Its molecular docking with active components of Platycodonis Radix-Armeniacae Semen Amarum was carried out by using Systems Dock Web Site network server. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis on the therapeutic targets of Platycodonis Radix-Armeniacae Semen Amarum were performed with the Database for Annotation, Visualization Andintegrated Discovery (DAVID). Result:Fourteen active components of Platycodonis Radix-Armeniacae Semen Amarum were screened out, including acacetin, cis-dihydroquercetin, spinasterol, licochalcone B, and luteolin, et al. One hundred and three therapeutic targets were screened out, including nitric oxide synthase 2 (NOS2), prostaglandin endoperoxide synthase 1 (PTGS1), androgen receptor (AR), prostaglandin endoperoxide synthase 2 (PTGS2), and dipeptidyl peptidase 4 (DPP4), et al. Identified signaling pathways mainly involved prostate cancer signaling pathway, small cell lung cancer signaling pathway, hepatitis B signaling pathway and T cell receptor signaling pathway. Conclusion:The possible molecular mechanism of Platycodonis Radix-Armeniacae Semen Amarum for the treatment of lung carcinoma was explored in this study based on network pharmacology, providing the direction for subsequent research.

Mechanism of Erzhiwan on Hepatocellular Carcinoma Based on Network Pharmacology / 中国实验方剂学杂志

Objective::To explore the possible mechanisms of Erzhiwan in the treatment of hepatocellular carcinoma (HCC) based on the network pharmacology. Method::The candidate active components and targets of Ligustri Lucidi Fructus and Ecliptae Herba were obtained through retrieval of the traditional Chinese medicine (TCM) systems pharmacology database (TCMSP) and literatures. Through Uniprot database and the human genome database (GeneCards), the overlapping genes of Erzhiwan and hepatocellular carcinoma were collected. The " candidate active components-targets" network of Ligustri Lucidi Fructus and Ecliptae Herba was built with Cytoscape 3.6.0 software. Drug target proteins and disease targets were mapped, and Venn map was drawn by Omicshare database. Major targets interaction network was formed by using String database and " Generate style from statistics" tool in Cytoscape 3.6.0 software. Molecular docking with active components was carried out by Systems Dock Web Site. The Gene Ontology (GO) classification enrichment analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for the targets were carried out in DAVID database. Result::Totally 21 active components, including beta-sitosterol, quercetin, luteolin, demethylwedelolactone, kaempferol, and 151 targets, including tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), N-terminal kinase (JUN), proto-oncogene (c-MYC), matrix metalloproteinase-9 (MMP-9) of Erzhiwan were collected. Erzhiwan exerts its effects on HCC mainly by acting on signal pathways, including Hepatitis B, TNF, Phosphatidylinositol-3-kinase (PI3K/Akt), tumor suppressor gene p53 and Toll-like receptor. Conclusion::Based on the methodology of network pharmacology, this study preliminarily predicted the major targets and pathways of Erzhiwan in the treatment of HCC, providing a direction for further studies.

Small-cell carcinoma of the prostate: A report of 2 cases and review of the literature / 中华男科学杂志

Objective@#To investigate the clinicopathological characteristics and improve the clinical treatment of prostatic small-cell carcinoma (PSCC).@*METHODS@#We reported 2 cases of PSCC derived from prostate cancer after treated by androgen blockade and prostate electrotomy and reviewed the relevant literature.@*RESULTS@#Two patients with PSA elevation were diagnosed with prostate cancer by prostatic puncture biopsy and treated by maximum androgen blockade, which reduced their total PSA to the normal level. Later, due to difficult urination, they both underwent prostate electrotomy, followed by chemotherapy or radiotherapy for PSCC confirmed by postoperative pathology. Nevertheless, they died at 8 to 9 months after the discovery of PSCC.@*CONCLUSIONS@#PSCC can derive from prostate cancer after treatment, which may be attributed to the pathological mutation induced by long-term endocrine therapy. PSCC is more malignant than prostate cancer, and its prognosis is poor.

Simultaneous Determination of Four Alkaloid Components in Xiangbin Decoction by UPLC-QqQ-MS/MS / 中国实验方剂学杂志

Objective: To establish a quantitative method for the simultaneous determination of arecaine,arecoline,norisoboldine and boldine in Xiangbin decoction by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-QqQ-MS/MS), and compare the variation of their contents between single and mixed decoctions. Method: The separation was carried on a Waters ACQUITY UPLC BEH Shield RP18(2.1 mm×150 mm,1.7 μm)column, with 0.1%formic acid solution-acetonitrile as mobile phase for gradient elution. The flow rate was 0.2 mL·min-1, and the column temperature was 30℃. The quantitative MRM transitions of the four components were m/z 142.10/44.11 for arecaine,m/z 156.20/44.07 for arecoline,m/z 314.29/265.12 for norisoboldine and m/z 328.13/265.10 for boldine. The determination was performed in multiple reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source under positive mode. Result: The linear ranges of arecaine,arecoline,norisoboldine and boldine were 0.479 0-57.48,0.976 0-78.08,0.812 0-64.96, 0.091 2-18.24 μg·L-1,respectively. The average recoveries of the above compounds ranged from 93.73%to 104.34%, with RSD (n=6) of less than 5%. The contents of arecoline,arecoline,norbibeldine and boltinine in Xiangbin decoction were (90.07±1.26),(445.27±12.39),(742.35±38.39),(38.50±3.33) μg·g-1,which were significantly lower than the contents in Linderae Radix and Arecae Semen. Conclusion: The method is rapid,sensitive,accurate and reproducible,and suitable for the simultaneous determination of multiple components in Xiangbin decoction,so as to provide a basis for the quality control of Xiangbin decoction. The compatibility of Xiangbin decoction has a significant effect on the dissolution contents of arecaine,arecoline,norisoboldine and boldine.

Study on influence of OVA specific immunotherapy on IL-23/Th17 axis of a mouse asthma model and its mech-anism / 局解手术学杂志

Objective To establish a mouse model of immune treatment of asthma through subcutaneous injection with high dose of ovalbumin( OVA) in the abdomen and investigate the role of IL-23/Th17 axis response in its mechanism. Methods With a random number table method, 18 female BALB/c mice were divided into 3 groups( normal control group,asthma group and asthmatic immune tolerance model group) ,with 6 mice in each group. The mice in the asthmatic immune tolerance model group were sensitized with 10 μg OVA by intraperito-neal injection in the abdomen on day 0 and day 7. The mice in the model group induced immune tolerance with 1 mg OVA by subcutaneous injection in the abdomen every day for a week(day 21 to day 27). Both the model group and asthma control group were challenged with 1％OVA on day 35 to day 41. The mice in the normal control group were challenged with the equal amount of saline. On the 50th day,each group were sforzando challenged once with 10％ OVA. The airway reactivity was detected at 24 after the last challenge. The enhanced pause( Penh) was measured to evaluate the airway responsiveness with a lung functional instrument. Bronchoalveolar lavage fluid( BALF) was collected to count the total cells and eosinophils,and the cytological studies were conducted. The OVA-specific IgE in peripheral blood,and the IL-5,IFN-γIL-23,IL-10 in BALF were detected by enzyme-linked immunosorbent assay(ELISA). The lung tissue was obtained to perform histological analysis by HE staining. The percentagea of Treg and Th17 cells in spleen and lung tissue were calculated by the flow cytometry( FCM) . Then the expression of transcription factors was detected by q-PCR. Results For the asthmatic immune tolerance model group, the airway respon-siveness,the cell count of eosinophilic granulocytes in BALF,the levels of IL-23 and OVA-specific IgE in the serum were significantly lower than the asthma group and the difference is of statistical significance(P<0. 05),while the difference in the IFN-γlevel in BALF compared with the asthma group is of no statistical significance(P<0. 05). The transcription factor of lung tissue detected with q-PCR showed Foxp3 in the asthmatic immune tolerance model group was significantly higher than the asthma group,while RORγt was significantly lower than the asthma group(P>0. 05) and the differences were of statistical significance(P<0. 05). Conclusion Large dose of OVA specific immuno-therapy can alleviate the chronic inflammatory response of asthmatic mice, and the decrease of Th17 cells associated with the expression of IL-23 was decreased. The mechanism may be related to the correction of the lung Il-23 /Th17 axis.

Inhibitory effect of polyphyllin Ⅰ on the proliferation of prostate cancer PC3 cells via ERK1/2/P65/DNMT1 and its molecular mechanism / 中华男科学杂志

<p><b>Objective</b>To explore the inhibitory effect of polyphyllin Ⅰ (PPⅠ) on the proliferation of castration-resistant prostate cancer PC3 cells and its molecular mechanism.</p><p><b>METHODS</b>We cultured human prostate cancer PC3 cells in vitro and treated them with PPⅠ at the concentrations of 0 (blank group), 0.4, 0.8, 1.2, 1.6, 2.0, and 2.4 μmol/L for 24, 48, and 72 hours, respectively. Then we detected the proliferation of the cells by MTT assay, measured their apoptosis by flow cytometry, and determined the expressions of p-ERK1/2, ERK1/2, NF-κB/p65 and DNMT1 proteins as well as the level of NF-κB/p65 in the cells additionally treated with the ERK1/2 inhibitor SP600125 by Western blot.</p><p><b>RESULTS</b>Compared with the blank control group, the PPⅠ-treated PC3 cells showed a concentration- and time-dependent reduction of the survival rate (1.00 ± 0.00 vs 0.85 ± 0.05, P < 0.01) at 0.4 μmol/L after 48 hours of intervention, concentration-dependent early apoptosis at 0.8 μmol/L (4.83 ± 0.95 vs 13.83 ± 2.97, P < 0.01), time-dependent increase of the expressions of p-ERK1/2 (1.00 ± 0.00 vs 1.73 ± 0.17, P < 0.01) and ERK1/2 (1.00 ± 0.00 vs 1.36 ± 0.12, P < 0.01) at 2 hours, and concentration-dependent decrease of the expressions of NF-κB/p65 and DNMT1 at 1.2 μmol/L (1.00 ± 0.00 vs 0.78 ± 0.10 and 0.63 ± 0.06, P < 0.01) and 1.6 μmol/L (1.00 ± 0.00 vs 0.67 ± 0.11 and 0.52 ± 0.09, P<0.01). Inhibition of ERK1/2 phosphorylation with PD98059 markedly reversed PPⅠ-induced decrease of the NF-κB/p65 expression as compared with that in the PPⅠ group (0.86 ± 0.18 vs 0.43 ± 0.09, P < 0.05).</p><p><b>CONCLUSIONS</b>PPⅠ induces the early apoptosis and suppresses the proliferation of PC3 cells, probably by activating the ERK1/2 pathway and inhibiting the expressions of the NF-κB/p65 and DNMT1 proteins.</p>

Effect of cadmium on mitochondrial function of hematopoietic stem cells in mouse bone marrow / 军事医学

Objective To explore the effect of cadmium chloride on mitochondrial function of hematopoietic stem cells in mouse bone marrow .Methods After being quarantined for one week , male Kunming mice weighted 20 ±2 g were randomly divided into three groups: control group , low dose cadmium-exposure group and high dose cadmium-exposure group.Mice in low dose and high dose cadmium-exposure groups were exposed to cadmium chloride solution at a dose of 7.5, 15 mg/kg body mass while those in control group were given an equal volume of distilled water through gavage administration every Monday , Wednesday and Friday for six consecutive weeks before cells in mouse bone marrow were collected at the 8th week.Mitochondrial membrane potential and ROS levels of mouse hematopoietic stem cells were detected using a flow cytometry .Results Compared with control group , the gain of body weight was significantly suppressed in cadmium-exposure group (P<0.01).Compared with control group, mitochondrial ROS levels of hematopoietic stem cells significantly increased in cadmium-exposure group and was dose-related(P<0.05,P<0.01). Besides, mitochondrial membrane potential of hematopoietic stem cells decreased in cadmium -exposure group compared with control group and was dose-related(P<0.05,P<0.01).Conclusion Cadmium exposure can lead to dose-related mitochondrial dysfunction of hematopoietic stem cells via oxidative damage in Kunming mice .

Three dimensional organoids-based evaluation for hepatotoxicity of the susceptible compound in Polygonum multiflorum Thunb / 药学学报

In this study, the three dimensional(3D)organoid culture system was established by liquid overlay method, and applied as an effective model to evaluate the hepatic injury of susceptible compounds in Polygonum multiflorum Thunb. Compared with the ordinary two dimensional(2D)culture of liver cells, the albumin expression of L02 cells and HepG2 cells were increased by 2.5 and 6.7 times in the 3D organoid culture system, respectively. After the cultivation of 21 days, urea generation levels of 3D culture were increased by 8.3 and 15.5 times. More importantly, HepG2 cells were more suitable to development of organoids than L02 cells. The gene expressions of phase I and II drug metabolism enzymes of HepG2 cells cultured as 3D organoids were significantly increased than that in 2D culture, such as the fold changes of CYP2C9 was up to 381.9, CYP3A4 to 87.0, CYP2D6 to 312.6. In addition, drug transporter relative genes were also up-regulated. The results demonstrated that the liver synthesis and metabolic function of the 3D model were better than that of the 2D cultured hepatocytes. The results of hepatotoxicity evaluation showed this developed model can be used to assess the hepatotoxicity of acetaminophen and other positive control drugs, which were considered with defined hepatotoxicity. On the 3D culture model, the IC50 value of repeated drug dose administration was significantly lower than that of single dose administration. However, the IC50 of 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside(cis-SG), which is the susceptible compound in Polygonum multiflorum Thunb., could not be detected in 2D cultured model. With the treatment of a single dose administration in organ 3D culture model, the IC50 of cis-SG was 1.9 times than that of cyclosporine A, and the IC50 of 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside(trans-SG)was 4.1 times than cis-SG. The hepatotoxicity results of cis-SG and trans-SG on the 3D cultures were similar to in vivo toxicity results obtained in previous work. On organ 3D culture model, the IC50 of cis-SG with repeat of administration decreased compared with that with single dose administration, suggesting that long-term medication may increase the risk of liver injury. In summary, the 3D organoid culture system can be used for a long period to preserve the capacity of liver synthesis and metabolism. The organoids were a model suitable for evaluation of mechanism of the drugs with low toxicity.

miR-206 inhibits renal cell cancer growth by targeting GAK / 华中科技大学学报（医学）（英德文版）

Renal cell cancer (RCC) remains one of the most lethal types of cancer in adults. MicroRNAs (miRNAs) play key roles in the pathogenesis of RCC. The role of miR-206 in RCC has not been fully understood. The purpose of this study was to examine the role of miR-206 in the regulation of proliferation and metastasis of RCC and the possible mechanism. miR-206 expression was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in RCC cell lines (786-O and OS-RC-2 cells) and clinical samples. MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] method, colony formation and transwell assay were used to detect the tumor-suppressing ability of miR-206 in RCC. Luciferase assay was performed to verify the precise target of miR-206. The results showed that the expression of miR-206 was significantly down-regulated in RCC tissues and cells. The expression level of cyclin G-associated kinase (GAK), a master regulator of tumor proliferation and metastasis, was up-regulated with the decrease in miR-206 in RCC tissues as well as RCC cell lines. In addition, the miR-206 inhibitor promoted the proliferation, migration and invasion of 786-O and OS-RC-2 cells. Bioinformatics combined with luciferase and Western blot assays revealed that miR-206 inhibited the expression of GAK. Moreover, miR-206 regulates RCC cell growth partly through targeting GAK. Our study indicated that miR-206 functions as a tumor suppressor in regulating the proliferation, migration and invasion of RCC by directly targeting GAK, and it holds promises as a potential therapeutic target for RCC.

Changes of the mitochondrial DNA copy number and the antioxidant system in the PBMC of hepatocellular carcinoma / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.</p><p><b>METHODS</b>The Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.</p><p><b>RESULTS</b>The copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).</p><p><b>CONCLUSION</b>There was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.</p>

Staging Based Strategies and Practice for Prostate Cancer / 中国中西医结合杂志

Authors raised that staging based strategies and practice of integrative medicine (IM) by combining syndrome typing and disease identification, and choosing suitable measures in accordance with different persons and seasonal conditions after more than ten years' clinical practice and researches. Radical operation as prior (as evil eliminating) and strengthening vital qi in perioerative period are best strategy for promoting rapid rehabilitation of early stage prostate cancer patients. Strengthening body resistance to eliminate evil was used in treating advanced prostate cancer patients. For example, a comprehensive treatment program for hormone-dependent patients was combined with endocrinotherapy and Chinese herbs for synergisic efficacy-enhancing actions. In this way, these patients' quality of life (QOL) were improved and time to castration resistant prostate cancer (CRPC) was delayed, even some patients were clinically cured. There are lack of effective medicines and methods for CRPC patients. Greatly tonifying original qi is mainly used for improving their clinical symptoms and prolonging survivals. Practice has proved staging based strategies and practice of IM has favorable advantages in treating prostate cancer, especially showing prospect in prolonging survival and postponing progression of advanced prostate cancer patients. Besides, it also could provide beneficial considerations and inspiration for combination of syndrome typing and disease identification.

Cluster Analysis of Medication Laws for Treating Coronary Heart Disease by Distinguished Veteran Doctors of Traditional Chinese Medicine / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To provide inspiration and ideas for clinical treatment of coronary heart disease (CHD) by data mining technology based frequency analysis and cluster analysis of medical records, prescriptions and herbs in treating CHD by distinguished veteran doctors of traditional Chinese Dedicine (TCM).</p><p><b>METHODS</b>Totally 386 medical cases were retrieved from Wanfang Data, Chinese Scientific Journals Database (VIP medical information resources system, China National Knowledge Infrastructure (CNKI), and Typical Collections of Medical Cases by Contemporary Distinguished Veteran Doctors of Traditional Chinese Medicine. They input into database trimmed after unified standard. Medication laws of CHD by distinguished veteran doctors of TCM were analyzed using frequency analysis and cluster analysis, and so on.</p><p><b>RESULTS</b>Distinguished veteran doctors of TCM frequently used top ten herbs in treatment of C D as Salvia miltiorrhiz , Ligusticum wallichii, Trichosanthes kirilowi, Pinellia ternat, Angelica sinensis, Poria coco stragalu , Panax ginseng, Allium macrostemon, and Radix Ophiopogonis. Cluster analysis summarized that there were 16 herb pairs commonly used, 7drug assemblies consisting of 3 herbs and 5 drug assemblies consisting of multiple herbs.</p><p><b>CONCLUSIONS</b>Distinguished veteran doctors of TCM mainly used herbs assemblies capable for invigorating Pi to resolve phlegm, and promoting qi and activating blood circulation in treating CHD. Meanwhile, they concurrently used herbs combination of nourishing Xin and tranquilization, and regulating yin and yang.</p>

Kidney-tonifying Chinese medicine for male infertility: A systematic review of randomized controlled trials / 中华男科学杂志

<p><b>OBJECTIVE</b>To systematically evaluate the efficacy and safety of kidney-tonifying traditional Chinese medicine in the treatment of male infertility.</p><p><b>METHODS</b>Based on the principles and methods of Cochrane systematic reviews, we searched CNKI, VIP, and Wanfang databases from inception to December 2012 for randomized controlled clinical trials addressing the treatment of male infertility with kidney-tonifying traditional Chinese medicine. According to the inclusion and exclusion criteria and retrieval strategies, we extracted the data, evaluated the quality of the included literature, and conducted meta-analysis using the RevMan 5. 2 software.</p><p><b>RESULTS</b>Twenty trials involving 2,272 patients were included, and the sample size of each study was from 60 to 270 cases. All the studies were graded as of poor quality, with Jadad scores of no more than 3 points. The results of meta-analysis showed that the total effectiveness rate of traditional Chinese medicine versus Western medicine on male infertility was RR = 1.71, 95% CI 1.19-2.47, and that of Chinese-Western combined therapy versus Western medicine was RR = 1.15, 95% CI 1.01-1.30. Both traditional Chinese medicine and Chinese-Western combined therapy showed a significantly better total effectiveness than Western medicine alone in improving the pregnancy rate without serious adverse reactions.</p><p><b>CONCLUSION</b>Due to the poor methodological quality and high heterogeneity of the included studies, the evidence for the efficacy and safety of kidney-tonifying traditional Chinese drugs in the treatment of male infertility is of but limited value, and further validation is needed by more high-quality studies.</p>

An open air research study of blast-induced traumatic brain injury to goats / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>We once reported blast-induced traumatic brain injury (bTBI) in confined space. Here, bTBI was studied again on goats in the open air using 3.0 kg trinitrotoluene.</p><p><b>METHODS</b>The goats were placed at 2, 4, 6 and 8 m far from explosion center. Trinitrotoluene (TNT) was used as the source of the blast wave and the pressure at each distance was recorded. The systemic physiology, electroencephalogram, serum level of S-100 beta, and neuron specific enolase (NSE) were determined pre and post the exposure. Neuroanatomy and neuropathology were observed 4 h after the exposure.</p><p><b>RESULTS</b>Simple blast waveforms were recorded with parameters of 702.8 kPa-0.442 ms, 148.4 kPa-2.503 ms, 73.9 kPa-3.233 ms, and 41.9 kPa-5.898 ms at 2, 4, 6 and 8 m respectively. Encephalic blast overpressure was on the first time recorded in the literature by us at 104.2 kPa-0.60 ms at 2 m, where mortality and burn rate were 44% and 44%. Gross examination showed that bTBI was mainly manifested as congestive expansion of blood vessels and subarachnoid hemorrhage, which had a total incidence of 25% and 19% in 36 goats. Microscopical observation found that the main pathohistological changes were enlarged perivascular space (21/36, 58%), small hemorrhages (9/36, 25%), vascular dilatation and congestion (8/36, 22%), and less subarachnoid hemorrhage (2/36, 6%). After explosion, serum levels of S-100b and NSE were elevated, and EEG changed into slow frequency with declined amplitude. The results indicated that severity and incidence of bTBI is related to the intensity of blast overpressure.</p><p><b>CONCLUSION</b>Blast wave can pass through the skull to directly injure brain tissue.</p>

Effect of Chinese herbs for stasis removing and collaterals dredging upon angiotensin-converting enzyme 2-angiotensin-(1-7)-mas axis in the renal cortex of diabetic nephropathy rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Chinese herbs for stasis removing and collaterals dredging (CHSRCD) upon angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas axis in the renal cortex of diabetic nephropathy rats.</p><p><b>METHODS</b>Totally 89 male Sprague-Dawley rats were randomly divided into the blank control group (C group, n=22), the high-glucose high-fat control group (H group, n=10), and the streptozotocin (STZ)-injecting group (n=57). The diabetes rat model (n=50) was induced by feeding high-glucose high-fat diet in combination with intraperitoneal injection of STZ, which were further divided into the model group (M group, n=24), the irbesartan group (I group, n=13), and the CHSRCD (Z group, n=13). Rats in I and Z groups were intragastrically fed with suspension of irbesartan and CHSRCD, once daily for 16 weeks. Equal volume of drinking water was administrated to rats in the rest groups. Blood glucose and 24 h urine protein quantitation were tested at four time points. And the mRNA expression of ACE2 and Mas at various time points was detected by Real-time PCR, immunohistochemical assay, and Western blot. Quantitative analyses of ACE2 and Mas protein expression were performed at the end of week 16.</p><p><b>RESULTS</b>Compared with the C group, blood glucose increased in the H and M groups (P < 0.01). It was higher in the H group (P < 0. 01). 24 h urine protein quantitation at different time points increased in the M group, and it was higher than that in the H group (P < 0.05). Compared with the M group, 24 h urine protein quantitation decreased at the end of week 8 in the I group, and at the end of week 8 and 16 in the Z group (P < 0.05). It was lower in the Z group than in the I group at the end of week 16 (P < 0.05). Compared with the C and H groups, the expression of ACE2 mRNA in the renal cortex was lower in the M group at the end of week 16 (P < 0.01). Compared with the M group, it was higher in the Z group (P < 0. 01). There was no statistical difference in the expressions of Mas mRNA at the end of week 16 between the C group and the M group (P > 0.05). It was lower in the M group than in the H group (P < 0.05). It was higher in the Z group than in the M group (P < 0.05), and higher than in the I group (P < 0.05). The expression of ACE2 and Mas protein in the M group decreased as time went by. The expression quantitation of ACE2 and Mas protein at the end of week 16 was lower in the M group than in the C group (P < 0.05). Compared with the M group, ACE2 expression of the Z group and Mas of the I and Z groups increased more significantly (P < 0. 05).</p><p><b>CONCLUSION</b>CHSRCD could play a role in renal protection for diabetic nephropathy rats by up-regulating the mRNA and protein expression of ACE2 and Mas, promoting the ACE2-Ang-(1-7)-Mas axis, and lowering urinary protein.</p>

Efficacy observation of cervical spondylosis treated with acupuncture at three lines of cervical Jiaji (EX-B 2) / 中国针灸

<p><b>OBJECTIVE</b>To compare the difference in the clinical efficacy on cervical spondylosis between acupuncture at three lines of cervical Jiaji (EX-B 2) and oral administration of jingfukang granules.</p><p><b>METHODS</b>Three hundred cases of cervical spondylosis were divided into an acupuncture group and a medication group, 150 cases in each one. In the acupuncture group, according to the different types of cervical spondylosis, acupuncture was applied at three lines of cervical Jiaji (EX-B 2), once a day. In the medication group, jingfukang granules were prescribed for oral administration, one bag each time, three times a day. The treatment of ten days made one session in the two groups and two sessions were required totally. Before and after two sessions of treatment, the clinical assessment scale for cervical spondylosis (CASCS) was adopted to evaluate the score of subjective symptoms, clinical physical signs and adaptability as well as the total score in the patients of the two groups and the efficacy was compared.</p><p><b>RESULTS</b>The patients' symptoms and physical signs were alleviated, the adaptability was improved and the score of each item and the total score were increased in the two groups after treatment (all P<0.01). The improvements in the acupuncture group were better than those in the medication group (all P<0.01). The curative and markedly effective rate was 90.7% (136/150) in the acupuncture group, better than 66.0% (99/150) in the medication group (P<0.01).</p><p><b>CONCLUSION</b>Acupuncture at three lines of cervical Jiaji (EX-B 2) achieves the significant clinical efficacy on cervical spondylosis. This therapy is superior to relieving symptoms and physical signs and recovering adaptability as compared with jingfukang granules.</p>