ObjectiveTo facilitate the establishment of fully automatic clinical diagnostic nucleic acid assay,a study was carried out for developing a fast and convenient method of extracting highly purified viral nucleic acid from human blood with magnetic beads.Method Five KHB (Shanghai Kehua Biological Engineering Company Limited) internal strong positive HBV serum samples were used in the process,for the method development,screening and selecting the magnetic beads based on magnetism,suspension power and extraction effects of HBV DNA,the optimization of the selected key components of extraction reagents were performed,which including the quantity of magnetic beads( 1 - 1.75 μl/ml),the buffer pH of lysis (4 -9),washing conditions with or without alcohol,elution for proliferation with or without magnetic beads,and etc.The evaluation for the extraction method was carried out including linearity,precision,sensitivity,specificity,anti-interference and resistance to contamination.The magnetic beads method was compared with domestic mature boiling method in a quantitative test with 100 HBV serum samples,the magnetic beads method was also compared with the interntional Gold standard Roche COBAS Tagman test method ( Roche method) in a quantitative test with 50 HBV serum samples and the results were statistically compared for their linear regression.ResultsOptimization experiment result showed superparamagnetic beads A was the best among the candidates ( C,B,D) in terms of their physical properties and extraction effects of DNA.The optimum condition of the assay were as follow:magnetic beads best used at 1.25 μl/ml,buffer pH 7 - 8 of lysis,ethanol was eliminated from washing buffer,proliferation can be carried out with magnetic beads.The parallel comparison experiments with five KHB internal strong positive HBV serum sample and gradient diluted serum sample between beads method and boiling method were carried out.The results showed that the linear range of beads method was 4 × 101 -4 × 108 IU/ml and that of boiling method was 4 × 102 -4 × 108 IU/ml.The sensitivity for beads method was 30 IU/ml and that of boiling method was 100 IU/ml.The precision CV of beads method was 6.0％ and boiling method was 11.9％.The false positive rate was 0 for beads method vs 3.1％ for boiling method.The anti-interference ability experiment showed that when sample contained sodium heparin concentration exceed 62.5 U/ml, there was significant drop of HBV DNA detection with boiling method and no HBV DNA was detected when sodium heparin concentration in the sample reached 1000 U/ml.However,sample sodium heparin concentration had no effect on HBV DNA detection with beads method.The comparison study between beads method and boiling method was carried out by use of 100 clinical HBV positive serum samples.The result showed that the sensitivity for beads method was 86％ vs boiling method 74％.The correlation between the two method was R2 =0.948,P ＜ 0.05.The comparison study between beads method and Roche method was carried use of 50 clinical HBV DNA positive serum samples and the result showed that sensitivity for beads method was 95％ (49/50) vs Roche method was 100％ (50/50).The correlation was R2 =0.963,P ＜ 0.05.ConclusionsThe established method for automatic nucleic acid extraction with magnetic bead displayed characteristic of high yield nucleic acid extraction,broader testing range,accurate quantification and convenient in operation.This can be one choice for nucleic acid extraction and applicable for clinical automatic diagnostic system.( Chin J Lab Med,2012,35:843 -850 )

Objective To investigate the effect of anti-epileptic soup on the levds of interleukin-2 (IL-2) and interleukin-6 (IL-6) in pentylenetetrazol-induced rats and its mechanism.Methods Healthy adult male SD rats were injected intraperitoneally with pentylenetetrazol kindling 35.0mg/kg to induce a rat epilepsy model.After successful construction of the epilepsy model,the rats were divided into four the groups:blank group,model group,anti -epileptic soup group and sodium valproate (VPA) group.All of the treatments were performed three times a day continuously for 28 days.The rats in the anti-epileptic soup group were given 6ml of anti-epileptic soup.The rats in the VPA group were given 400mg/kg of sodium valproate by garage.The rats in blank group and model group received 6ml of normal saline.After treatment,the seizures level was recorded by using the Diehl six point grading scale,and the expression of serum IL-2 and IL-6 were detected by the enzyme linked immunoassay (ELISA).Results The seizure level of the model group was (4.23 ± 0.43),while that of the anti-epileptic group was (3.47 ± 0.39),indicated that the anti-epileptic soup can reduce the seizures level.The expressions of IL-2 (4.18 ± 0.41)pg/mL and IL-6 (350.0 ± 82.5) pg/mL of the model group were higher than those of the blank group [IL-2 (3.20 ± 0.36) pg/mL,IL-6 (260.2 ± 69.0) pg/mL,P ＜ 0.01],while the expressions of IL-2 (3.48 ± 0.44) pg/mL and IL-6 (287.5 ±74.6) pg/mL of the anti-epileptic group were lower compared with the control group (P ＜0.01).No significant difference was found between the anti-epileptic group and VPA group IL-2 (3.35 ± 0.56) pg/mL and IL-6 (253.8 ± 59.8) pg/mL(P ＞ 0.05).Conclusion The anti-epileptic soup is effective for treating epilepsy in rats,the possible mechanism may be related with downregulating the expression of IL-2 and IL-6.

OBJECTIVETo study the chemical constituents from the antioxidant fraction of Neo-Taraxacum siphonathum.

METHODVarious chromatographic techniques were used to isolate and purify the constituents. The structures were elucidated on the basis of chemical evidence and spectral analysis.

RESULTTen compounds were isolated and identified from Neo-T. siphonathum, caffeic acid (1), chlorogenic acid (2), quercetin (3), luteolin (4), quercetin-3-O-beta-D-glucopyranoside (5), quercetin-3-O-alpha-D-arabinofuranoside (6), quercetin-3-O-alpha-D-arabinopyranoside (7), luteolin-7-O-beta-D-glucopyranoside (8), beta-sitosterol (9) and daucosterol (10).

CONCLUSIONCompounds 1-10 were isolated from this plant for the first time.

Caffeic Acids ; chemistry ; isolation & purification ; Chlorogenic Acid ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Luteolin ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Quercetin ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification ; Taraxacum ; chemistry

Objective: To analyze the application of accelerated rehabilitation surgery in elderly patients with gastric cancer surgery and its influence on inflammation and nutritional indicators. Methods: From October 2017 to October 2018, 80 elderly patients with gastric cancer who underwent gastrectomy in Shaoxing People's Hospital were selected.According to random number table method, they were randomly divided into traditional control group and ERAS group, with 40 cases in each group.The traditional control group was treated by traditional perioperative treatment + operation, while ERAS group was treated with ERAS perioperative treatment + operation.The recovery and complications, inflammation and nutritional changes before operation, 1 day after operation and 3 days after operation, and the improvement of quality of life after operation were compared and analyzed between the two groups. Results: In the ERAS group, the first exhaust time[(2.3±0.8)d] and defecation time[(2.5±0.4)d]were shorter than those in the traditional control group[(3.5±0.5)d and (3.7±0.6)d], and the incidence rate of complications (7.5%) was lower than that in the traditional control group (35.0%), the differences were statistically significant (t=8.72, 10.52, χ²=9.04, all P<0.05). On the first day after operation, the serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6(IL-6) in the ERAS group were (28.3±4.9)μg/L and (23.1±5.8)μg/L, respectively, which were lower than those in the traditional control group[(39.8±6.7)μg/L and (34.5±8.1)μg/L](t=8.76, 7.24, all P<0.05). The serum TNF-α and IL-6 levels in the ERAS group on the 3rd day after operation were (18.4±6.2)μg/L and (18.3±3.2)μg/L, respectively, which were lower than those in the traditional control group[(26.2±5.1)μg/L and (26.5±4.8)μg/L](t=6.14, 8.99, all P<0.05). The serum levels of albumin (Alb) and prealbumin (PRE) in the ERAS group on the first day after operation were (31.1±0.7)g/L and (161.3±7.2) mg/L, respectively, which were lower than those in the traditional control group[(30.2±0.9)g/L and (154.3±4.8)mg/L](t=4.99, 5.12, all P<0.05). The serum levels of Alb and PRE in the ERAS group on the 3rd day after operation were (33.4±0.5)g/L and (172.1±5.0)mg/L, respectively, which were lower than those in the traditional control group[(31.9±0.8)g/L and (165.3±3.2)mg/L](t=10.06, 7.24, all P<0.05). The WHOQOL-BREF score of the ERAS group[(91.4±6.5)points]was higher than that of the traditional control group[(80.3±7.8)points](P<0.05). Conclusion Accelerated rehabilitation surgery has good effect for elderly gastric cancer surgery.It can maintain the inflammation and nutritional status of patients after operation.It is worthy of clinical reference.

Objective:To investigate the clinical value of three-dimensional arterial spin marker imaging(3D-ASL) for evaluating collateral circulation compensatory status in patients with cerebral infarction.Methods:A total of 24 patients with cerebral infarction who were treated at Wenzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang University of Chinese Medicine from September 2017 to January 2019 were examined by 1.5-T high-field MR scanner.The changes of cerebral blood flow(CBF) between the infarcted area and the contralateral normal area were compared when the labeled delay time(PLD)=1.5s and PLD=2.5s, and the patients were divided into the group with strong collateral circulation compensation and the group with weak collateral circulation compensation according to the difference in lesion area measured by 3D-ASL(PLD=1.5s and 2.5s). The changes of national institutes of health stroke scale(NIHSS) score and Bathel index at admission and 15 days after admission were compared.Results:3D-ASL measurement of PLD=1.5s showed that the CBF value in the infarcted area was (33.70±20.83)mL/(100g×min), which was significantly lower than that in the contralateral normal area(PLD=1.5s)[(49.93±13.13)mL/(100g×min)]( t=3.229, P<0.05). When PLD=2.5s, ASL measurement results showed that the CBF value in the infarcted area increased significantly[(58.26±23.50)mL/(100g×min) vs.(33.70±20.83)mL/(100g×min)]( t=3.831, P<0.05), and the CBF value in the contralateral normal area also increased significantly[(68.29±14.03)mL/(100g×min) vs.(49.93±13.13)mL/(100g×min)]( t=4.681, P<0.05). At this time, the CBF value in the infarcted area was still significantly lower than that in the contralateral normal area[(58.26±23.50)mL/(100g×min) vs.(68.29±14.03)mL/(100g×min)]( t=1.795, P<0.05). On the day of admission, the BI index of patients in the weak collateral circulation compensation group(12 cases) was significantly higher than that in the strong collateral circulation compensation group(12 cases)[(90.42±10.50) vs.(67.92±27.57)]( t=2.642, P<0.05), and the NIHSS score was significantly lower than that in the strong collateral circulation compensation group[(1.25±1.01) vs.(3.83±3.62)]( t=2.378, P<0.05). After 15 d of admission, there were no significant differences in BI index and NIHSS score between the two groups(all P>0.05). Conclusion:3D-ASL with different delay time can effectively and intuitively reflect collateral circulation compensation of patients with cerebral infarction, and it has very important reference value for the evaluation of patients' disease and the formulation of clinical treatment plan.

Animals ; Blood-Brain Barrier ; metabolism ; Ferritins ; metabolism ; Horses ; Mice ; Receptors, Transferrin ; metabolism ; Spleen ; chemistry