Objective To compare the oil yield of celery seeds and the contents of 3-n-butylphthalide and the total phthalocyanine lactones of celery seed oil extracted by different methods. Methods Three routine extraction methods involving organic solvent extraction, Soxhlet extraction, steam distillation extraction, as well as subcritical extraction method and supercritical fluid extraction method were used to extract the celery seed oil. The contents of 3-n-butylphthalide and total phthalocyanine lactones were respectively detected by high performance liquid chromatography(HPLC) and ultraviolet visible spectrophotometry. Results The ranges of oil yield and the contents of 3-n-butylphthalide and total phthalocyanine lactones of celery seed oil extracted by different methods were 0.30%-20.02%, 1.40%-10.13%, 4.74%-17.65%, respectively, indicating obvious differences. Conclusion With R134a and butane as the solvents, the subcritical extraction method is better than other extraction methods for the extraction of 3-n-butylphthalide. With dimethyl ether as the solvent, the subcritical extraction method is the best for the extraction of total phthalocyanine lactones.

Objective Fluctuation of glucose levels is more likely to cause oxidative stress which contributes to the development of insulin resistance through activating c-Jun N-terminal kinase (JNK).Such antio xidants as vitamin C or vitamin E do not appear very helpful.Radix Astragali (RA) is an herbal medicine with antioxidative ability.The study was to explore whether RA would lower the risk of insulin resistance in diabetic rats.Methods Diabetic rats received RA,insulin or both RA and insulin after diabetes were induced in male Wistar rats.Serum levels of interleukin 6 (IL-6) and tumor necrosis factor α (TNFα),advanced glycation endproducts(AGEs) levels in kidney,the expression of insulin receptor (IR),insulin receptor substrate 1 (IRS-1),p38 mitogen-activated protein kinase (MAPK),p-p38 MAPK,p-JNK,p-IRS-1 Ser307,and p-IRS-1 Tyr612 in skeletal muscles were determined.Results Compared to diabetic rats treated with insulin,the diabetic rats treated with both insulin and RA demonstrated significantly lower levels of IL-6,TNF-α and AGEs (P ＜ 0.05),significantly lower activation of p-p38 MAPK and JNK (P ＜0.05),significantly higher expressions of IRS-1 (P ＜0.05),p-IRS-1 Tyr612,and significantly lower expression of p-IRS-1 Ser307 (P ＜ 0.05).Conclusions RA can lower the risk of insulin resistance through fighting oxidative stress in diabetic rats.

Objective To evaluate the effects of mild hypothermia performed at different times on the levels of glutamate, Bcl-2 and Bax during global cerebral ischemia-reperfsusion (I/R) in rats. Methods Twenty-four male SD rats weighing 230-270 g were randomly divided into 4 groups according to the time at which mild hypothermia was performed ( n =6 each): group A, B, C and D. Global cerebral ischemia was produced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and occlusion of bilateral common carotid arteries) in the 4 groups. In group B, C and D, the nasopharyngeal temperature was reduced to 32.5-33.5 ℃ and maintained for 1 h. Ischemia was performed after termination of cooling in group B. While ischemia was performed, cooling was started in group C. While reperfusion was performed, cooling was started in group D. Rewarming was started after termination of cooling in group B, C and D. Samples of dialysate in hippocampal CA1 area were collected every 10 min during 100 min reperfusion for determination of glutamate concentrations by high-performance capillary electrophoresis with laser-induced fluorescence detection ( HPCE-LIF). The brain tissues were taken at 3 h of reperfusion for determination of the expression of Bax and Bcl-2 in hippocampal CA1 area, and the ratio of Bcl-2/Bax was calculated. Results Compared with group A, the glutamate concentrations were significantly decreased, the Bcl-2 expression was up-regulated, the Bax expression was down-regulated and the ratio of Bcl-2/Bax was increased in the other thee groups ( P ＜ 0.05). Compared with group B, the Bcl-2 expression was significantly down-regulated, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group C ( P ＜0.05). The glutamate concentrations were significantly increased, the Bax expression was up-regulated and the ratio of Bcl-2/Bax was decreased in group D compared with group C ( P ＜ 0.05 ). Conclusion The earlier cooling is performed, the better the cerebral protective effect is during global cerebral I/R in rats.

Objective To study the effects of electro-acupuncturing points of Yangming meridians on hemiplegia.Methods45 early hemiplegic patients were divided into Groups A,B and C(18 cases in each group).Physical therapy was performed in each group,those in Group B were electro-acupunctured at points of Yangming meridians,while those in Group C at the points according to balancing muscular tension.They were assessed with Fugl-Meyer Assessment and Ashworth Spasm Rating before and 2,4 weeks after treatment.Results2 weeks after treatment,motor function and muscular tension were significantly improved in all the groups,while the efficacy in Groups B and C was superior to Group A(P<0.05).4 weeks after treatment,the Ashworth score of lower extremity in Group B was superior to Groups C and A(P<0.05).ConclusionElectro-acupuncturing at points of Yangming meridians can improve the motor function especially muscular tension in hemiplegic patients,which may be helpful for flaccid paralysis,but adverse for spastic paralysis.

AIM:To observe the effect of plumbagin on the mRNA and protein expression of nicotinamide ade -nine dinucleotidephosphate oxidase 4 ( Nox4 ) , reactive oxygen species ( ROS ) level and protein expression of α-smooth muscle actin (α-SMA) in the HSC-LX2 cells stimulated with transforming growth factor β1 (TGF-β1) in vitro.METH-ODS:HSC-LX2 cells were cultured in vitro and divided into blank group, model group, high-, medium-and low-dose (2, 1.5 and 1 μmol/L) plumbagin groups .After incubated with each drug for 72 h, the mRNA expression of Nox4 was detec-ted by RT-PCR.ROS levels were tested by in situ loading probe method.The protein contents of Nox4 and α-SMA were measured by Western blot .RESULTS:Compared with model group , after treated with plumbagin for 72 h, the mRNA ex-pression of Nox4, ROS level and α-SMA protein were significantly decreased in high-and medium-dose plumbagin groups (P<0.01).CONCLUSION:Plumbagin inhibits the activation of HSC-LX2 cells via decreasing the expression of Nox4, thus decreasing ROS levels .

Objective To analysis of the detection result of amniotic fluid chromosome which in NIPT high-risk pregnant women.Methods Amniotic fluid cells via amniotic cavity puncture were cultured and analyzed,the chromosome karyotypes were observed.Results The highest positive predictive value of NIPT was for trisomy 21(85.00%),then trisomy 18(75.00%),sex chromosome abnormalities(68.00%),other chromosome abnormalities(41.67%),trisomy 13 (25.00%).Conclusion The highest accuracy of NIPT was shown in detection of Down''s syndrome by NIPT.NIPT was screening test which is effective and noninvasive in prenatal diagnosis.Amniotic fluid Chromosomal karyotype analysis was the gold standard in the diagnosis of fetal chromosomal disease.

Huaihua medical College has inquired into the questionnaire from more 500 students who are clinical doctors and graduate nurses and 50 teachs who are clinical teaching hospitals,finding that the results have indicated the significance,the necessity and the actual problems of clinical technical ability practice teaching,when they are in school by their firsthand experience,which are very helpful to shortening scheduled time to practice,improving clinical basic skills,adjusting to medical environment as soon as possible,to ensure practice teaching quality and also have notable effects on continuing reinforcing building the clinical technical ability training centre and improving the quality of clinical technical teaching ability at the same time.

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

Objective To evaluate the diagnostic value of virtual touch imaging quantification (VTIQ) technique in the differential diagnosis between benign and malignant breast lesions.Methods From September to December 2015, the imaging data of 115 breast lesions in 100 patients confirmed by pathology on conventional ultrasound (2DUS) and VTIQ were retrospectively analyzed. The breast nodules were examined by 2DUS firstly and then the lesions were classified by breast imaging reporting and data system (BI-RADS). The maximum, minimum and average of shear wave velocity (SWV) values were obtained from multiple SWV measurement under the VTIQ speed mode. According to the pathology results, receiver operating characteristic (ROC) curve were plotted to determine the most accurate SWV value and the cut-off value for differential diagnosis. And the diagnosis efficiency was compared between 2DUS and VTIQ.Results There were 38 malignant nodules and 77 benign nodules in 115 breast nodules. BI-RADS grading≥4b level was set as the malignant nodules and grading≤ 4a level as the benign nodules. The SWVmax, SWVmin, and SWVmean on VTIQ of benign and malignant breast nodules were (5.11±1.61) m/s, (2.90±0.86) m/s, (3.73±1.02) m/s, and (7.22±0.94) m/s, (4.33±1.31) m/s, (5.73±1.11) m/s. There were significant differences between malignant and benign breast nodules in SWVmax, SWVmin, and SWVmean on VTIQ (t=-7.442,-7.047,-9.356, allP<0.001). Based on the area under curve of ROC, the SWVmean value in the nodule was the best value in comparison with other SWV values. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of diagnose malignant lesion conducted by 2DUS were 89.5％, 85.7％, 87.0％, 0.63 and 0.84, respectively. The cut-off value of VTIQ mean was 4.34 m/s. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value for the diagnosis of malignant lesion conducted by VTIQ were 89.5％, 85.7％, 87.0％, 0.76 and 0.95, respectively. The diagnosis efficiency of VTIQ was better than 2DUS.Conclusion The study proved that the VTIQ SWVmean is the best parameter for differential diagnosis and VTIQ plays an important role in differential diagnosis of breast nodules.

BACKGROUNDThe expressive level of glucose-regulated protein 78 (GRP78) is elevated and correlated with resistance of chemotherapy drugs in breast cancer cell. However, little is known about the relationship between its expression and drug resistance in non-small cell lung cancer (NSCLC). The aim of this study was to explore the relationship between drug resistance and the expression of GRP78 in NSCLC.

METHODSDrug sensitivity test was used to detect the resistance to 8 chemotherapy drugs in 52 NSCLC fresh surgical samples by methylthiazoletrazolium (MTT), and expression of GRP78 was detected by immunohistochemistry method. Spearman correlation assay was used to investigate the correlation between the GRP78 expression and drug resistance.

RESULTSThe resistance rates to paclitaxel (PTX), adriamycin (ADM), carboplatin (CBP), topotecan (TPT), navelbine (NVB), vincristine (VCR), cisplatin (DDP) and etoposide (VP-16) of the 52 samples were 42.31%, 57.69%, 63.46%, 65.38%, 67.31%, 73.08%, 78.85%, 90.38%, respectively. Fourteen cases showed the complete resistance to the total 8 chemotherapy drugs. Furthermore, the expression of GRP78 was stronger in poorly differentiated cancer as compared with the well and moderately differentiated cancer (P < 0.05), so as in stage II and III cancer than in stage I cancer (P < 0.05). Spearman correlation assay showed that there was a correlation between the chemotherapeutics resistance to ADM, VP-16, VCR, TPT and the expression of GRP78 in NSCLC (P < 0.05).

CONCLUSIONSIt is feasible to detect the drug sensitivity to chemotherapy for tumor cells by MTT method. The results of chemosensitivity assay in vitro are indicative of clinical drug administration in NSCLC. The detection of GRP78 isalso indicative of the resistance to chemotherapy drugs and the differentiation and the clinical stage in NSCLC.