Objective To explore the reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells by tetrandrine ( Tet) and to clarify its molecular mechanism.Methods LOVO/5-Fu cells were treated for 48 h with Tet.Drug sensitivity was measured by MTT.The cell cycle, apoptosis of cells and expression of P-glycoprotein (P-gp) were determined by flow cytometry assay.Expression of MDR1 mRNA was detected by real-time quantitative PCR (real-time PCR).P-gp expression was detected by Western blot.Results After LOVO/5-Fu cells were treated for 48 h with Tet, the IC50 of 5-Fu decreased to ( 4.15 ± 0.31 ) μg/ml ( P ＜ 0.05 ) ; and the apoptotic rate increased to (3.44% ± 0.28% ) ( P ＜ 0.05) ; the expression of MDR1 mRNA reduced to (570 ± 85) (P ＜ 0.05 ).Conclusions Tetrandrine reverses MDR1 gene-mediated multidrug resistance in human colon carcinoma LOVO/5-Fu cells possibly by inhibiting the expression of MDR1, decreasing the expression of P-gp, thus enhancing the sensitivity of LOVO/5-Fu cells to 5-fluorouracil.

Colorectal neoplasms always present with thickness of the intestinal wall or a soft tissue mass in the enteric cavity. Multi-slice computed tomography (MSCT) with high spatial resolution and advanced post-processing techniques can demonstrate the above signs of the tumor, and the invasive signs of adjacent structures and lymph node metastasis. Combined with three dimensional reformation images, MSCT shows a higher sensitivity than that of double contrast barium enema and electronic colonoscope. MSCT is promising in the diagnosis and preoperative evaluation of colorectal neoplasms.

The art of warm disease of TCM not only has abundant clinical experience,but also has a profound theoretical thinking,and it starts with its unique concept category and logical system,and riches and enriches,updates and develops on the basic of this,and goes along the road of independent development,has a distinct characteristics.This paper preliminarily studies on wei qi ying and xue of the art of warm disease of TCM from meaning,development of history,nature and action,et al.And it provides references of ideas and methodologies for the development and innovation of the art of warm disease of TCM.

Objective To evaluate the safety of temporary cardiac pacemaker during perioperative period of laparoscopic cholecystectomy (LC) in patients with bradyarrhythmia. Methods Temporary cardiac pacemakers were installed preoperatively in 34 elderly patients with bradyarrhythmia for LC. On operation the ultrasonic harmonic scalpel, instead of electrotome, was used to perform dissection and coagulation. Results LC was completed successfully in all the 34 patients without surgery-related complications or perioperative serious cardiovascular diseases. Conclusions Utilization of temporary cardiac pacemaker and ultrasonic harmonic scalpel may considerably enhance the perioperative safety and reduce the incidence of cardiovascular complications in elderly patients with bradyarrhythmia.

Objective To compare the efficacy of XELOX and FOLFOX4 in the treatment of locally advanced unresectable gastric cancer. Methods The clinical data of 72 patients with gastric cancer who were admitted to the Shandong Provincial Hospital from July 2006 to October 2009 were prospectively analyzed. Of all the patients, 3 lost follow-up, and 69 patients with locally advanced unresectable gastric cancer were randomly divided into XELOX group ( n = 36 ) and FOLFOX4 group ( n = 33 ) according to the random number table.All patients received chemotherapy for six weeks. The efficacy of the two regimens were evaluated by the multidiscipline team six weeks later. The cell cycle of patients with complete or partial remission and received surgical treatment was detected by flow cytometry. All data were analyzed using the Pearson chi-square test, Levene test or t test. Results The curative rates of XELOX and FOLFOX4 were 53% (19/36) and 52% (17/33), respectively,with no significant difference between the two groups ( x2= 0. 01 , P ＞ 0. 05 ). The incidences of nausea and vomiting, phlebitis and hand-foot syndrome were 25% (9/36), 6% (2/36) and 19% (7/36) in the xELOX group, and 55% ( 18/33), 39% (13/33) and 3% (1/33) in the FOLFOX4 group, respectively, with significant difference between the two groups ( x2 = 6.31, 11.59, 4.53, P ＜ 0.05 ). Nineteen patients in the XELOX group and 17 patients in the FOLFOX4 group received surgical resection of the gastric cancer, and no complications such as anastomotic leakage and hemorrhage occurred postoperatively. In the XELOX group, the s-phase fraction (SPF),proliferation index (PI) and G2/M of the gastric cancer cells were 5.89% ± 0.79%, 9.22% ± 1.99% and 5.19% ± 1. 66% after neoadjuvant chemotherapy, which were significantly lower than 6.76% ± 1.21%, 10.44% ±2.12% and 6. 04% ± 0. 57% before neoadjuvant chemotherapy, while the ratio of gastric cancer cells in the G0/G1 phase after neoadjuvant chemotherapy was 90.39% ±4.78%, which was significantly higher than 87.54%±6.34% before neoadjuvant chemotherapy (x2 =3.61, 2.52, 2. 15, 2.91, P ＜0.05). In the FOLFOX4group, the SPF, PI and G2/M of the gastric cancer cells were 6.09% ± 0.96%, 10.65 % ± 2.47% and 4.88% ±0.87% after neoadjuvant chemotherapy, which were significantly lower than 7.15% ± 1.45%, 11.87% ± 2.33%and 5.67% ± 1.03% before neoadjuvant chemotherapy, while the ratio of gastric cancer cells in the G0/G1 phase after neoadjuvant chemotherapy was 91.45% ± 5.22%, which was significantly higher than 88.01% ± 4.23%before neoadjuvant chemotherapy ( x2 = 3.50, 2.06, 3.37, 2.94, P ＜ 0.05 ). There was a significant difference in PI between XELOX group and FOLFOX4 group after neoadjuvant chemotherapy ( x2 = 2.66, P ＜ 0.05 ).Conclusions XELOX and FOLFOX4 are safe and effective in the treatment of locally advanced unresectable gastric cancer, and they can significantly restrain the proliferation of gastric cancer cells. XELOX regimen is more effective than FOLFOX4 regimen.

Objective To describe the clinical characteristics of one child with primary hyperoxaluria types Ⅲ, and to analyze the potential mutant genes in his family.Methods AGXT, GRHPR and HOGA1 genes were analyzed by direct sequencing analysis in this family.One hundred unrelated healthy subjects were also analyzed as controls.Results The child had early onset of symptoms (0.8 year).His principal clinical manifestation included nephrolithiasis and obstructive nephropathy, however his nephrocalcinosis was mild.And he presented high urine oxalate, high urine calcium, and lower citrate levels.Two novel heterozygous mutations in HOGA1 were identified (compound heterozygous), one mutation was a 2-bp substitution at the last position in exon 6 and the first position of intron 6 respectively (c.834_834 + 1GG ＞ TT);another was a guanine to adenine substitution of the last nucleotide of exon 6 (c.834G ＞ A).Both of these variants found in this study probably acted as splicing mutations.Direct sequencing analysis failed to find these mutations in 100 unrelated healthy subjects.In addition, a SNP (c.715G ＞ A, p.V239I) was found in this family.There were no mutations detected in AGXT and GRHPR.Conclusions Two novel mutations are identified probably in association with PH Ⅲ.This is the first description and investigation on mutant gene analysis of PHⅢ in Asia.

Objective To compare the reversal effects of short hairpin RNA (shRNA) interference and tetrandrine on multidrug resistance (MDR) of human colorectal cancer cell line LoVo/5-fluorouracil(5-FU ).Methods An eukaryotic expression plasmid of shRNA targeting MDR1 was constructed and transfected into human colorectal cancer cell line LoVo/5-FU (transfection group).LoVo/5-FU was also pretreated with tetrandrine (tetrandrine group).Drug sensitivity was detected by methyl thiazolyltetrazolium colorimetric method.Cell cycle,apoptosis of cells and positive expression rate of P-glycoprotein (P-gp) were determined by flow cytometry assay.The expressions of MDR1 mRNA and P-gp were detected by real-time polymerase chain reaction and Western blot,respectively.All data were analyzed by analysis of variance and SNK-q test.Results (1)Drug sensitivity:the 50％ concentration of inhibition(IC50)of the control group,tetrandrine group and transfection group were (7.3 ± 0.3),(4.4 ±0.7) and (2.4 ±0.4) mmol/L,respectively,a significant difference between the 3 groups was found(F =65.27,P ＜ 0.05 ).There was a significant difference in the IC50 between the tetrandrine group and the transfection group (q =6.67,P ＜ 0.05 ).(2) Changes of cell cycle:the proportion of cells in the G1 phase and S phase of the control group,tetrandrine group and transfection group were 38.13％ ± 3.75％,51.36％ ± 2.76％,59.24％±4.31％ and 20.46％±2.23％,14.32％± 1.91％,9.40％± 1.65％,respectively,a significant difference between the 3 groups was found(F =25.23,24.37,P ＜ 0.05 ).There were significant differences in the proportion of cells in the G1 phase and S phase between the tetrandrine group and the transfection group(q =3.67,4.35,P ＜ 0.05 ).(3) Cell apoptosis:the cell apoptotic rates of the control group,tetrandrine group and transfection group were 1.32％ ± 0.47％,3.24％ ± 0.26％,5.88％ ±- 0.44％,respectively,a significant difference between the 3 groups was found(F =99.26,P ＜ 0.05 ).There was a significant difference in the cell apoptotic rate between the tetrandrine group and transfection group(q =11.48,P ＜ 0.05 ).(4)The expression of P-gp:the positive expression rates of P-gp of the control group,tetrandrine group and transfection group were 96.9％ ± 2.3％,61.6％ ± 4.9％,76.6％ ± 3.6％,respectively,a significant difference between the 3 groups was found(F =67.83,P ＜ 0.05 ).There was a significant difference in the positive expression rate of P-gp between the tetrandrine group and transfection group (q =6.97,P ＜ 0.05 ).(5)The mRNA expression of MDR1:the mRNA expressions of MDR1 of the control group,tetrandrine group and transfection group were 1462 ±161,570 ±85,233 ± 81,respectively,a significant difference between the 3 groups was found(F =90.59,P ＜ 0.05 ).There was a significant difference in the mRNA expression of MDR1 between the tetrandrine group and transfection group (q =5.12,P ＜ 0.05 ).Conclusions MDR1 shRNA and tetrandrine could reverse M DR1 gene-mediated m.ultidrug resistance in human colon cancer cell line LoVo/5-FU,but the effect of MDR1 shRNA is better than that of tetrandrine.MDR1 shRNA and tetrandrine might take effect by inhibiting P-gp expression and down-regulating mRNA expression of MDR1.

Biopsy, Needle ; methods ; Bronchoscopy ; Child ; Humans ; Pediatrics ; methods

Objective:To explore the diagnostic value of mammography and ultrasonography(US)in diagnosis of breast disease with microcalcifications.Methods:Seventy-eight cases with breast microcalcifications were detected by mammography.The detection rate of breast microcalcification at US was analyzed.And the sensitivity,specificity and the accuracy of US in diagnosis breast disease with microcalcifications was compared with these of mammography.Results:The detection rate of breast microcalcification was 66.7%,And US depicted more malignant(87.5%)than benign microcalcification(33.3%).The sensitivity specificity and accuracy of US was 68.7%、83.3%、 74.3%respectively,and the sensitivity specificity and accuracy of mammography was 72.9%、 73.3%、75.6%respectively,there were no significant difference between the two methods.The sensitivity,specificity and accuracy of the combined the two methods was 89.6%,90.0%,92.3% respectively.Conclusion:US can effectively detect and identify the breast microcalicications.The combination can improve the assessment of breast disease with microcalicications,and has a significant clinical practical value in diagnosis of early breast cancer.

Objective To describe and analyze the clinical characters of patients with FRG from 7 Chinese families.Then analyze and identify their mutations in SGLT2 gene,and explore the association of genotype and phenotype.Methods Quantitative test for 24-hour urine glucose and other laboratory tests were carried out among 7 probands (14 patients in all) and their family members from 7 pedigrees (totaling 23 subjects).All coding regions,including intron-exon boundaries,were analyzed using PCR followed by direct sequence analysis.Results Five novel mutations in SLC5A2 gene were identified in this investigation,including four missense mutations (A Serine to Glycine at position 335 (c.1003A＞G,p.S335G),a Glutamine to Arginine at position 448 (c.1343A ＞ G,p.Q448R),an alanine to proline at position 474 (p.A474P,c.1420G ＞ C) and a glycine to aspartic acid at position 580 (c.1739G ＞ A,p.G580D) and a deletion in intron 7 (c.886(-10_-31)del).By the minigene studies using the pSPL3 plasmids,we confirmed the deletion c.886(-10_-31)del as a splicing mutation.In this study,the mutation c.886(-10_-31)del accounted for about 43％ of the total alleles (12/28).These patients with compound heterozygous or homozygous mutations manifested middle degree or severe glycosuria (Quantitative test for 24-hour urine glucose:10.56-50.68 g/1.73 m2),however those with heterozygous variants presented with mild to moderate glycosuria (Quantitative test for 24-hour urine glucose ≤ 2.45 g/1.73 m2).This fits co-dominant inheritance pattern.Conclusions Five novel mutations which may be related to FRG are found in this study,and c.886(-10-31) del may be a high frequency mutation in Chinese patients.