Objective To investigate the effects of shRNA produced by PCR on the suppression of the expression of exo- or endogenous genes. Methods The specific primers were designed, with which the shGFP-RNA and shFN-RNA were produced by PCR. The shGFP-RNA was transfected into 293T cell lines together with pEGFP plasmid, then the cells were detected by laser confocal microscopy 48h later. The shFN-RNA was transfected into rat mesenteric cell lines, then the cells were collected 48h later and the total RNA was extracted, which was reversely transcripted to cDNA. Then the expression level of FN mRNA was examined with real-time PCR, and the expression level of FN protein was examined with Western blot analysis. Results The results of laser confocal microscopy indicated that the EGFP could be successfully suppressed by shGFP-RNA produced by PCR; the results of real-time PCR and Western blot analysis revealed that FN expression level of the cells transfected with shFN-RNA was down regulated, and the level was much lower than those tansfected with independent shRNA (P

Objective To assess the clinical value of dual-energy CT (DECT) Volume software in quantitative analysis of urate crystals.Methods The DECT data of 60 gout patients based on the American College of Rheumatology diagnostic criteria were analyzed retrospectively.The volumes of urate crystals were quantitatively analyzed by using Volume software with two senior radiologists.The results were statistically analyzed.Results Seventy-two joints of 60 gout patients were scanned by DECT.40 of 43 joints had urate crystals in foot and ankle with the average volume of (0.621±0.742) cm3;18 of 19 joints had urate crystals in knee with the average volume of (0.842±1.086) cm3;10 of 10 joints had urate crystals in hand and wrist with the average volume of (0.796±0.583) cm3.There was no statistical difference for volume measurement between two doctors (P>0.05).The volumes of urate crystals in 4 patients with regular medication were reduced.Conclusion Volume software of DECT can quantitatively analyze urate crystals with a good repeatability, which has high application value in clinical diagnosis and treatment monitoring of gout.

Objective To explore the mechanism of protecting cells from ischemic reperfusion injury by constructing specific small interference RNA (siRNA) to inhibit Na+-H+exchanger-1 (NHE-1) expression in human renal tubular epithelial cell (HKC). Methods The siRNA was designed and synthesized based on human NHE-1 complete sequence,and was transfected into HKC.The irrespective siRNA transfected group was used as control.The cells were treated with 10 μmol/L antimyein A to induce ischemia and anoxyaemia environment.NHE-1expression was examined by RT-PCR and Western blot.The intraeellular pH (pHi),Ca2+ or Na+ concentrations were detected by BCECF/AM,Fluo-3/AM and SBFI-AM,respectively,combining with laser eonfocal assay system.Nucleic morphology was determined by Hoechst 33342.Cellular apoptosis was examined by Annexin V/PI staining and flow eytometry.Fluorescent probe JC-1 was used to detect the change of mitechondrial transmembrane potential. Results The specific siRNA could efficiently inhibit NHE-1 expression in HKC.Compared with the irrespective siRNA transfected group,the mRNA and protein expression of NHE-1 was significantly down-regulated in NHE-1 siRNA transfeeted group (all P＜0.05).After treatment with antimyein A,the mRNA and protein expression of NHE-1 was significantly up-regulated in both groups,however,it was less than that in irrespective siBNA transfected group.At the same time,the ratio of apoptosis decreased (8.9% +2.9% vs 18.8%±3.2% , 17.4%±3.6% ,P＜0.05) and mitochondrial transmembrane potential rose significantly in NHE-1 siRNA transfected group as compared to irrespective siRNA transfected group and antimycin A group.The intracellular Na+,H+ and Ca2+concentrations increased in NHE-1 siRNA transfected group treated with antimyein A,but their levels were lower than those in irrespective siRNA transfected group with the same treatment(P＜0.05). Conclusions The synthesized siBNA can inhibit the expression of NHE-1 and can protect HKC from isehemia reperfasion injury induced by antimyein A.The mechanism might be via suppressing the expression of NHE-1 to delay intracelluar Na+ accumulation,attenuate intracellular Ca2+ overloading,and inhibit the decrease of mitechondrion transmembrane potential and reduce cellular apoptosis.

Objective To establish analysis methods for fingerprint of Chuanxiong Rhizoma by HPLC. Methods Thermo C18 chromatographic column (4.6 mm×250 mm, 5 μm) was used with methanol-0.1% Formic acid in gradient elution. The flow rate was 1 mL/min, the detection wavelength was set at 323 nm, and the temperature was 25 ℃. The similarities of the 18 batches of samples were compared by similarity evaluation, cluster analysis and principal component analysis. Results Based on the fingerprints of 18 batches of Chuanxiong Rhizoma, 11 common peaks were identified, the similarities were almost greater than 0.9 among all batches. The samples were clustered into 3 categories. Conclusion The method is simple, steady and repeatable. It provides a basis for the quality control and evaluation of Chuanxiong Rhizoma.

Biosimilars are biological medicinal products that are highly similar to an already licensed reference product in terms of quality, safety, and efficacy. BAT1706 is being developed by Bio-Thera Solutions, Ltd. as a proposed biosimilar candidate to bevacizumab reference product (Avastin^®). Bevacizumab acts by specifically binding to vascular endothelial growth factor A (VEGF-A), and preventing the interaction of VEGF-A with its receptors on the surface of endothelial cells, then blocking the downstream signaling pathway mediated by ligand-receptor, and inhibiting endothelial angiogenesis, thus inhibiting tumor growth. Comprehensive analytical characterization studies incorporating orthogonal analytical techniques were performed to compare the in vitro functional activities of BAT1706 and Avastin^®. BAT1706 and Avastin^® showed highly similar binding activity to multiple VEGF-A isoforms and equivalent VEGF-A neutralizing activity, as well as inhibitory activity of VEGF receptor (VEGFR)-2 tyrosine kinase autophosphorylation. Both products exhibited similar binding of the Fcγ receptors and a lack of Fc-related effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Overall, the results demonstrate that BAT1706 and Avastin^® are highly similar in terms of in vitro functional activities.

Objective To study the clinical effect of adjacent flap on immediate breast repair in conserving surgery for upper inner quadrant breast cancer.Methods A total of 100 women with breast cancer treated in our hospital from August 2012 to August 2016 were selected as the study subjects.According to the different surgical methods,46 cases were divided into control group and 54 cases in observation group.The patients in the control group were treated with modified radical mastectomy for breast cancer.The patients in the observation group were treated with adjacent flap for immediate repair of breast defect in breast conserving surgery.The quality of life,the follow-up results and between the two groups were compared.The cosmetic effect of the observation group was evaluated.Results At 6 months after operation,the physiological function,affective function,physical pain,social function,physiological limitations,vitality,mental health and overall health score of the observation group were higher than those of the control group,the difference was statistically significant(P ＜ 0.05).There was no significant difference in local recurrence rate,distant metastasis rate,non-survival rate,loss of follow-up or mortality between the two groups (P ＞0.05).The excellent and good rate of breast cosmetic effect was 96.3％ after surgery in observation group.The satisfaction rate of observation group was 92.59％ (50/54),the control group was 45.46％ (21/46),the difference was statistically significant (x =26.582,P =0.000).Conclusion The clinical effect of repairing breast defect by using adjacent flap transfer is very effective in improving the quality of life and reducing the risk of local recurrence and distant metastasis.

OBJECTIVETo compare the diagnostic value of barium enema (BE), computed tomography (CT) and magnetic resonance imaging(MRI) in primary colorectal carcinoma.

METHODSA total of 64 patients with suspected colorectal carcinoma received BE (n=39), spiral CT (n=31) and MRI (n=42). The detective results were compared with the surgical results.

RESULTSAmong 64 patients, 54 cases were pathologically proved as colorectal carcinoma. The diagnostic sensitivity of BE,CT and MRI was 96.9% ,96.2% and 97.1% ,and the overall accuracy was 92.3% 83.9 % and 90.5% respectively. The overall accuracy of CT and MRI for tumor T staging was 73.1% and 82.9% respectively.

CONCLUSIONBE can be considered as a primary approach for diagnosing colorectal carcinoma, CT and MRI be necessary diagnostic approaches. Combined BE with MRI is the best choice for diagnosing of colorectal carcinoma.

Adult ; Aged ; Aged, 80 and over ; Barium Sulfate ; Colorectal Neoplasms ; diagnostic imaging ; pathology ; Enema ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm Staging ; Sensitivity and Specificity ; Tomography, Spiral Computed

OBJECTIVETo explore the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSCs) on injured hepatocytes mediated by paracrine mechanisms and to investigate the potential molecular mechanism of this action.

METHODSA contact-independent model of aberrant hepatic microenvironment was established by co-culturing BM-MSCs with D-galactosamine (D-GalN)-injured human L02 hepatic cells using a transwell assay platform. Secreted levels of insulin-like growth factor-1 (IGF-1) were measured by enzyme-linked immunosorbent assay of the co-culture supernatant. Expression of the IGF-1 receptor (IGF-1R) was assessed by Western blot. The effect of exogenous IGF-1 on proliferation of D-GalN-injured L02 cells was examined by MTT assay.

RESULTSUpon co-culture, BM-MSCs promoted proliferation of D-GalN-injured L02 cells in a contact-independent manner (absorbance values of at 24 h: 0.36+/-0.08, 48 h: 0.52+/-0.06, and 96 h: 0.68+/-0.06; vs. uninjured cells t = 2.493, 3.116, and 2.285, respectively; all P less than 0.05). Robust expression of IGF-1 was identified in the supernatants of co-cultures and was demonstrated to have been secreted mainly from BM-MSCs under the influence of D-GalN-injured L02 cells. Constitutive expression of IGF-1R was found in the D-GalN-injured L02 cells and blocking of IGF-1R by a neutralizing antibody significantly inhibited the paracrine pro-proliferative effect of co-cultured BM-MSCs at 24 h, 48 h, and 72 h (t = 2.909, 2.328, and 2.560, respectively; all P less than 0.05).

CONCLUSIONBM-MSC-derived IGF-1 plays an important role in the paracrine pro-proliferative effect on D-GalN-injured L02 hepatocytes by engaging with the constitutively expressed IGF-1R on L02 cells.

Bone Marrow Cells ; metabolism ; Cell Proliferation ; Cells, Cultured ; Hepatocytes ; cytology ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Mesenchymal Stromal Cells ; metabolism

OBJECTIVE: To determine the therapeutic effect of spirulina platensis in allergic rhinitis (AR). METHODS: Ovalbumin sensitized white rats used as AR animals were treated with spirulina platensis (SPP). At the end of the treatment, the differences in the behavior science were observed; the changes in the nasal mucosa and mast cell degranulation were studied pathologically; and the levels of serum histamine and total immunoglobulin (Ig) E were determined by enzyme-linked immune sorbent assay. RESULTS: The behavior science score of the SPP treatment group was lower than that of the negative control group (P < 0.01 ) ; inflammatory reaction of nasal mucosa in the SPP treatment group were remarkably relieved; the number of nasal mucosa mastocyte and mast cell degranulation in the SPP treatment group were lower than that of the negative control group (P <0.01 ). The levels of serum histamine and total IgE in the SPP treatment group were lower than that of the negative control group (P <0.01 ). It had no significant difference in the positive control group and the SPP treatment group and the blank control group (P > 0.05 ). CONCLUSION Spirulina platensis can prevent and treat AR in rats, which implies the possibility of using spirulina platensis for AR patients in the future.
Animals ; Eukaryota ; Male ; Ovalbumin ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic, Perennial ; chemically induced ; drug therapy

Objective To investigate the neurotoxic effects ofLDN-57444, a specific ubiquitin C-termiual hydrolase L1 (UCH-L1) inhibitor, on dopaminergic neurons and the possible mechanism. Methods The viability of SK-N-SH cells exposed to 5, 10, 25, 50, 75 or 100 μmol/L LDN-57444 for 24 h was assessed using MTT assay, and the cell apoptosis was detected with Hoechst staining. Western blot was performed to identify the expressions of UCH-L1 protein, ubiquitin monomer and polyubiquitinated proteins, and the activity of the ubiquitin-proteasome system (UPS) was evaluated with fluorometry. Results After exposure to UCH-LI inhibitor for 24 h, the cell process-like structures of SK-N-SH cells diminished, and the cell body shrank and became spherical. Exposure to LDN-57444 resulted in concentration-dependent reduction of the cell viability, and the reduction became statistically significant following the exposure to 50 μmol/L LDN-57444, as compared with that in the control group (P<0.05). The exposure also resulted in obvious cell apoptosis as shown by nuclear fragmentation and presence of the apoptotie bodies. Western blot detected no obvious changes in UCH-L1 protein expression but identified reduced ubiquitin monomer and increased polyubiquitinated protein expression in the cells. Fluorometry showed reduced activity of UPS in the exposed cells. Conclusion UCH-L1 inhibitor produces neurotoxicity to dopaminergie neurons and induces cell apoptosis possibly as the result of impaired UPS activity and intracellular accumulation of polyubiquitinated proteins following the exposure.