Objective:To study the change of microRNA during the early stage of high phosphorus in-duced vascular smooth muscle cell (VSMC)calcification and its related mechanism.Methods:The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol /L)for 7 d.The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content,real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules.Based on the cell calcification model,micro-RNA microarray array was applied to screen the profiles of microRNA expression in VSMC following high Pi stimulation for different periods (0,3 and 12 h).The array data were analyzed by TAMtool to explore the activated signaling pathway.Results:The calcium content of A7r5 cells induced by high Pi was in-creased 9.6 times high as cells without Pi treatment (P <0.05 ).VSMC contractile phenotype genes (SM-αactin,SM22)were down-regulated (P <0.05 ),while calcification-related genes (BMP2, MSX2,Runx2)were up-regulated (P <0.05)in VSMC stimulated by high Pi.The calcium nodules were obviously formed in cells after 7 d high Pi treatment.In microarray experiment,680 individual mi-croRNAs were detected in high Pi-treated VSMCs at different time points (0,3 and 12 h).Among these genes,miR-183,miR-664 and miR-9 * were increased whereas miR-542-5P,let-7f and miR-29a were decreased in time-dependent manners.Twenty-six kinds of signaling pathways,including cell apoptosis, differentiation and proliferation,were significantly activated.All these activated pathways were associated with calcification.Conclusion:This study implies that microRNA changed in high Pi-induced VSMCs may involve in the process of calcification.

AIM:To determine whether human peritoneal mesothelial cells express CD40. METHODS:Human peritoneal mesothelial cells (HPMC) were harvested from spent peritoneal dialysis effluent and maintained under defined in vitro conditions. Expression of CD40, CD40L and intercellular adhesion molecule-1 (ICAM-1) on HPMC under normal culture or stimulation with interferon-γ (IFN-γ), tumor necrosis factor-α(TNF-α), interleukin(IL)-1 and LPS were detected by FACS analysis. The relationship between CD40 expression and ICAM-1 expression on HPMC was analyzed.RESULTS:HPMC cultured in vitro expressed CD40 constitutively. The surface expression of CD40 was markedly up-regulated following stimulation with IFN-γ, but not with TNF-α, IL-1 and LPS. CD40L expression on HPMC was not detected. The expression of ICAM-1 on HPMC was significantly increased by stimulation with IFN-γ, TNF-α, IL-1, LPS, respectively; the most effective inducer on ICAM-1 expression was IFN-γ. The CD40 expression co-localizes with the expression of ICAM-1 and there was positive correlation between CD40 and ICAM-1 expression on HPMC. CONCLUSION:HPMC functionally expressed CD40.

AIM: To investigate the effect of 4.25%peritoneal dialysis solution (PDS) on CD40 expression in rat peritoneal mesothelial cells so as to reveal the potential mechanisms by which CD40-CD40 ligand (CD40L) interaction may be involved in the inflammation of peritoneal membrane. METHODS: Rat peritoneal mesothelial cells (MC) were harvested from the peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with 4.25%PDS or 4.25%PDS+IFN-? was detected by RT-PCR and FACS analyses. After activation of CD40 on MC with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. RESULTS: MC cultured in vitro expressed CD40 constitutively. 4.25%PDS markedly up-regulated the expression of CD40 mRNA and its protein. The expression of CD40 mRNA and its protein following stimulation with 4.25%PDS+IFN-? was significantly higher than 4.25%PDS alone. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with CD40mAb.CONCLUSIONS: MC functionally express CD40. The up-regulated CD40 expression on MC following stimulation with 4.25%PDS may play an important role in local peritoneal defense mechanisms and may be involved in the chronic inflammatory process of the peritoneum.

This study examined the dynamic characteristics of upper airway collapse at soft palate level in patients with obstructive sleep apnea/hypopnea syndrome (OSAHS) by using dynamic 3-Dimensional (3-D) CT imaging. A total of 41 male patients who presented with 2 of the following symptoms, i.e., daytime sleepiness and fatigue, frequent snoring, and apnea with witness, were diagnosed as having OSAHS. They underwent full-night polysomnography and then dynamic 3-D CT imaging of the upper airway during quiet breathing and in Muller's maneuver. The soft palate length (SPL), the minimal cross-sectional area of the retropalatal region (mXSA-RP), and the vertical distance from the hard palate to the upper posterior part of the hyoid (hhL) were compared between the two breathing states. These parameters, together with hard palate length (HPL), were also compared between mild/moderate and severe OSAHS groups. Association of these parameters with the severity of OSAHS [as reflected by apnea hypopnea index (AHI) and the lowest saturation of blood oxygen (LSaO(2))] was examined. The results showed that 31 patients had severe OSAHS, and 10 mild/moderate OSAHS. All the patients had airway obstruction at soft palate level. mXSA-RP was significantly decreased and SPL remarkably increased during Muller's maneuver as compared with the quiet breathing state. There were no significant differences in these airway parameters (except the position of the hyoid bone) between severe and mild/moderate OSAHS groups. And no significant correlation between these airway parameters and the severity of OSAHS was found. The position of hyoid was lower in the severe OSAHS group than in the mild/moderate OSAHS group. The patients in group with body mass index (BMI)≥26 had higher collapse ratio of mXSA-RP, greater neck circumference and smaller mXSA-RP in the Muller's maneuver than those in group with BMI<26 (P<0.05 for all). It was concluded that dynamic 3-D CT imaging could dynamically show the upper airway changes at soft palate level in OSAHS patients. All the OSAHS patients had airway obstruction of various degrees at soft palate level. But no correlation was observed between the airway change at soft palate level and the severity of OSAHS. The patients in group with BMI≥26 were more likely to develop airway obstruction at soft palate level than those with BMI<26.

The present paper outlined pesticide registration status for traditional Chinese medicines (TCMs) and summarized the characteristics of pesticide contamination in different regions of some widely used TCMs by retrieving last 10 years' literatures. At present, the problems of pesticide residues for TCM include less pesticide registrations, widespread high-residue organochlorine pesticides contamination, pesticide abuse, irregular GAP bases and imperfect pesticide limit standards, etc. According to the current situation, we should adopt some control measures to strengthen the quality control of TCMs so as to ensure the safety of TCMs and related products.
Animals ; China ; Drug Contamination ; prevention & control ; statistics & numerical data ; Humans ; Medicine, Chinese Traditional ; methods ; Pesticide Residues ; analysis ; Registries

OBJECTIVETo evaluate the effectiveness of manual therapy and traction for lumbar disc herniation and analyze the current status of this kind of randomized clinical trial (RCT).

METHODSDatabase of CNKI, VIP, WANFANG, PubMed and OVID were searched. Some relevant journals were manually retrieved. A total of 2 874 literatures on manual therapy and traction for lumbar disc herniation were collected, of which 17 articles met the inclusion criteria. The Jadad score scale was used to evaluate the quality,and RevMan5.0 was used for meta-analysis of literatures.

RESULTSThe results of the meta-analysis of all trials involved were as followed:the combined effect of the effective rate was RR = 1.10, 95% CI [1.06, 1.14], the combined effect of the cure rate was RR = 1.36, 95% CI [1.21,1.52], the combined effect of the VAS was RR = 1.37, 95% CI [1.28, 1.45], the combined effect of the JOA was RR = 4.75, 95% CI [4.40, 5.09].

CONCLUSIONThe overall quality of the current RCT researches about manual therapy for lumbar disc herniation was lower,and did not support the conclusion that manual therapy was more effective than traction for lumbar disc herniation.

Humans ; Intervertebral Disc Displacement ; surgery ; therapy ; Lumbar Vertebrae ; surgery ; Musculoskeletal Manipulations ; methods ; Randomized Controlled Trials as Topic ; Traction ; methods

OBJECTIVETo compare the differences in the clinical efficacy on Alzheimer's disease between acupuncture and medicine.

METHODSOne hundred and forty-one patients were randomized into an acupuncture group (72 cases) and a medicine group (69 cases). In the acupuncture group, the needling technique for benefiting qi, promoting blood circulation, regulating mind and improving intelligence was used at Shenting (GV 24), Baihui (GV 20), Fengchi (GB 20), Wangu (GB 12), Danzhong (CV 17), Zhangwan (CV 12), Qihai (CV 6), Xuehai (SP 10) and Zusanli (ST 36). The supplementary acupoints were selected according to the symptoms and physical signs. Acupuncture was given once a day and 6 treatments were required for a week. In the medicine group, the choline sterase inhibitor, donepezil (aricept) was prescribed for oral administration, 1 tablet (5 mg) each time, once every night. Four weeks later, the dose was increased to 2 tablets (10 mg) each time. In the two groups, the treatment of 4 weeks made one session and 4 sessions were required. The changes of scores before and after treatment in the minimum mental state examination (MMSE), the activity of daily living scale (ADL), Alzheimer's disease assessment scale-cognition (ADAS-cog) and the digit span (DS) were observed.

RESULTSAfter treatment, scores of MMSE and DS were increased as compared with those before treatment (both P < 0.05) and scores of ADL and ADAS-cog were reduced as compared with those before treatment. The score differences in MMSE, ADL, ADAS-cog and DS before and after treatment were significant in the two groups (all P < 0.01).

CONCLUSIONThe needling technique for benefiting qi, promoting blood circulation, regulating mind and improving intelligence significantly improves the overall function, cognition and activity of daily life in the patients of Alzheimer's disease and the efficacy is better than donepezil.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Aged ; Aged, 80 and over ; Alzheimer Disease ; psychology ; therapy ; Cognition ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

Cytometric bead array (CBA) is a new analytical technique, which can achieve real-time and rapid detection of targeted components in a small amount of sample. With many advantages of high throughput screening, high specificity and sensitivity, low cost, easy operation and good repeatability, this CBA technique has been widely used for the detection of various components in foods, agricultural products and environmental samples. Recently, it has got significant development in rapid detection of small molecules. This review briefly introduced the theory of CBA technique, summarized the application in the analysis of small molecules, such as mycotoxins, pesticide residues, shellfish toxins, and then prospected the application of trace small molecules detection in the complex matrices of traditional Chinese medicine and the development trend of it.
Drug Contamination ; High-Throughput Screening Assays ; instrumentation ; methods ; Immunoassay ; instrumentation ; methods ; Microspheres ; Pesticides ; analysis ; Toxins, Biological ; analysis

In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Gene Knockdown Techniques ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

Objective To establish CFPAC-1 cell lines deficient in CDC25B2 by recombinant lentivirus, and to investigate the role of this gene. Methods After CFPAC-1 cells were transduced with recombinant lentivirus producing CDC25B2 siRNA, stably transduced cells with green fluorescent protein were selected by flow cytometer. The mRNA and protein expression of CDC25B2 was examined by RT-PCR and Western blot analysis. The effect of the lentivirus on the cell proliferation, cell cycle, clone-forming, migration and invasion ability was analyzed by MTr method, flow cytometer, plate clone-forming assay and Transwell chamber method respectively. Results CDC25B2 siRNA knocked down CDC25B2 expression in CFPAC-1 cells significantly. The silencing efficiency of siRNA transduction by recombinant lentivirns was very high. Proliferation, cloneforming, migration and invasion ability of human pancreatic cancer cell line CFPAC-I were significantly in-creased, while cell cycle was not affected. Conclusion CDC25 B2 plays an important role in cell proliferation, clone-forming, migration and invasion of pancreatic cancer. This research provides experimental evidences for targeting CDC25B2 in gene therapy against pancreatic cancer.