Objective To establish determination method for total polyphenols in Guhong Injection, and provide reference for internal quality control of Guhong Injection.[Methods]Using the Folin-Ciocalteu colorimetric method, according to the consistent degree of the maximum absorption wavelengths of hydroxysafflor yel ow A and kaempferol relative to Guhong Injection, we selected the appropriate reference and color reaction condition to content determination.[Rsults]With hydroxysafflor yel ow A as the standard, the content of total polyphenols in 10 batches of samples were 9.94, 9.55, 9.75, 9.67, 9.84, 10.03, 9.81, 9.52, 9.88, 10.09mg·mL-1. The average recovery of total polyphenols was 98.11% and RSD was 1.68%(n=6). [Conclusion]The established Folin-Ciocalteu method is simple, accurate, sensitive, and is reliable for detecting total polyphenols in Guhong Injection.

BACKGROUND: Extraction of fungal DNA plays an important role in fungal genetic engineering and molecular biology research.The result of experiment is affected seriously by the efficiency of extracting DNA especially the quality of DNA. OBJECTIVE: To develop a method for extracting genomic DNA of pathogenic fungi and discuss the optimal combination of components in simple sequence repeat PCR (SSR-PCR) system. DESIGN, TIME AND SETTING: A comparative analysis of DNA extraction methods and an orthogonal experiment were conducted in the Mycology Research Lab of Department of Pathogenobiology in Norman Bethune College of Medicine, Jilin University from July 2004 to December 2006.MATERIALS: Clinical specimens were inoculated on Potato dextrose agar, Potato dextrose broth and Yeast extract peptone dextrose and cultivated under the temperature of 28 ℃ for 3-7 days, after which suspectable colonies were selected to be isolated and purified.METHODS: Three kinds of methods of extracting DNA(beading-salt fractionation method, CTAB method and Gene TLE~(TM) extraction method ) were compared in terms of their effects on DNA quality; Experiment was performed with orthogonal design to four factors (Taq DNA polymerase, template DNA, dNTP, primers) in three levels on the basis of L9 (3~4) orthogonal table, The appropriate annealing temperature and cycles were determined through PCR.MAIN OUTCOME MEASURES: The optimal reaction system determined according to the polymorphism and specificity of amplification of banding pattern.RESULTS: The objective fragments were all amplified by Gene TLE~(TM) extraction method, and the banding patterns obtained were clearer and brighter compared with the other two methods. The result of orthogonal experiment on SSR-PCR system showed that,according to the value of R, the significance of factors followed by ascending were template DNA (2.67), Taq DNA polymerase (2.00), dNTP (0.67) and primers (0.33). According to the value of ki, the optimal level of each factor combination was 30 mg/L template DNA, 1U Taq DNA polymerase, 150 μmol/L dNTP, 0.5 μmol/L primer. However, because primers were nonsignificant factors, which was presented by their small R value, we took A level of primer as 0.25 μmol/L. The best reaction condition was 55 ℃ annealing temperature and 35 cycles.CONCLUSION: The Gene TLE~(TM) method shows higher efficiency of extracting DNA and its operation is fast and simplel According to the results of orthogonat experiment, the optimal SSR-PCR system was 30 mg/L template DNA, 1U Taq DNA polymerase, 150 μmol/L dNTP and 0.25 μmol/L primer. The best reaction condition was 55℃ annealing temperature and 35 cycles.

Objective To develop the methods for using 0.015 cc pinpoint chambers, 0.007 cc miniature chambers and diode detector to measure Multi-leaf collimator (MLC) small field in IMRT.Methods MAX4000 and Unidos electrometers were connected with different types of small chambers and diode detectors.MLC shaped fields of10 cm×10 cm, 6 cm×6 cm, 4 cm×4 cm, 3 cm×3 cm, 2 cm× 2 cm were defined at 100 cm SSD.The field sizes for the Varian accelerator were defined by the tertiary MLC, while the secondary jaws were kept at 10 cm × 10 cm field, with the monitor units of 250 MU.Each field was measured three times to obtain the average value.The readings of all small fields were normalized to 10 cm × 10 cm field values for comparison of measured and published output factors.Results The relative deviations of the MLC small field output factors from the published outputs are 1.0％ , 1.7％ , 1.5％ and 2.4％, respectively, for Unidos electrometer connected with 0.015 cc pinpoint chamber;0.2％, 0.8％, 0.8％ and 1.4％, respectively, for Unidos electrometer connected with 0.007 cc miniature chamber;and 0.1％, 0.5％, 0.5％ and 0.9％, respectively, for MAX4000 electrometer connected with 0.007 cc miniature chamber.Conclusions The 0.015 cc chamber-measured MLC output factors for 3 cm × 3 cm and 2 cm × 2 cm fields are excellent.As required by IAEA, the relative deviations of the measured output factor from the published output factor are within ± 2％ for 2 cm × 2 cm fields and ± 3％ for larger fields.The results measured using 0.007 cc chamber are better than those measured using 0.015 cc chamber.The measured results using the diode detector, normalized to the 10 cm × 10 cm field, are consistent with the minimum requirements and excellent when being normalized to the 4 cm × 4 cm field.For dosimetric consideration, MLC small field output factor should be measured using small chamber and diode detector.The method is accurate and reliable, therefore, all measured output factors for MLC small fields should be input into radiation treatment plan system.

Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.

Objective:To explore the postoperative complications of different incisions of parotidectomy in benign parotid tumor and the impact on life quality.Methods:62 patients with benign parotid ttmor underwent improved parotidectomy in our hospital from January 2010 to January 2015 were selected and randomly divided into group A and group B.The patients in group A were using improved S incision,and the patients in group B were using postauricular concealing incision.Then the perioperative indexes,complications after surgery and the influences to life quality of 2 groups were observed and compared.Results:The surgery time,blood loss,postoperative suction drainage and hospital stays of 2 groups had no great differences (P＞0.05).The early and forward complication rate of group A was 29.03 ％ and 25.81％ respectively,of group B was 19.35 ％ and 12.90 ％ respectively.There were no differences between them (P＞0.05).The scores of pain and emotion after surgery of group A were getting better,and appearance,smell and chewing function was getting worse than before surgery with statistically significance (P＜0.05).The scores of pain and emotion after surgery of group B were getting better than before (P＜0.05).The scores of appearance and emotion of group A were worse than those of group B with statistically significance (P＜0.05).Conclusions:Using postauricular concealing incision can obtain good life quality and safety for the patients with benign parotid tumor,which is superior to improved S incision,worthy of clinical applications.

Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.

Objective To compare the effects of target systolic blood pressure on cardiovascular events in patients with hypertension and type 2 diabetes after antihypertensive drugs. Methods This study was a retrospective cohort study. Randomly selected from March 2014 to March 2017, 1563 patients with hypertension and type 2 diabetes diagnosed in the Department of Cardiology of the Tiemei Group General Hospital. There was no cardiovascular complications at admission. After admission, the population variables of each patient were recorded. Each patient was adjusted for blood pressure and blood glucose according to the doctor's advice after admission. After the hospital was discharged, the patient was followed up for blood pressure every week. The follow-up was 3-4 months. The average systolic blood pressure of each patient was recorded as the target systolic pressure. The patients who were followed up were divided into three groups according to the target systolic pressure value, namely ≤120 mmHg(1 mmHg=0.133 kPa) group,>120-130 mmHg group,>130-140 mmHg group. After that, it was changed to a weekly telephone follow-up of the nurses for a total of 3 years. The cardiovascular disease (CVD) occurred in these patients within 3 years were recorded. The CVD was used as the dependent variable, and the systolic blood pressure of each group was used as the independent variable to do COX. Proportional risk regression analysis, the hazard ratio (HR) of each group of concurrent CVD. Taking CVD as the dependent variable, HR of CVD with >130-140 mmHg group was taken as the reference value, and all population variables were used as independent variables, respectively, ≤120 mmHg group,>120-130 mmHg group and>130-140 mmHg group combined with CVD risk ratio. Results During the 3-year follow-up period, 1401 patients with blood pressure met: 104 patients with systolic blood pressure ≤120 mmHg, 543 patients with >120-130 mmHg, and 754 patients with >130-140 mmHg. The incidence of CVD in the three groups was 14.4％(15/104), 8.8％(48/543), 7.0％(53/754). HR of CVD were 1.66, 0.96, 0.53 in≤120 mmHg group,>120- 130 mmHg group,>130-140 mmHg group .Compared with >130-140 mmHg group, the risk ratio of CVD in all subgroups of≤120 mmHg group was statistically increased (HR>1, P<0.05). Compared with >130-140 mmHg, >120-130 mmHg group with 65-79 years old, combined with more than 3 groups of antihypertensive drugs, patients with diuretics showed a statistically significant increase in the risk ratio of CVD (HR>1, P <0.05). Conclusions The target systolic blood pressure of patients with hypertension and type 2 diabetes should be >130-140 mmHg. For patients aged 65-79 years, more than 3 groups of antihypertensive drugs are used. Patients with diuretics should have a target systolic blood pressure of >130-140 mmHg.

Objective To retrospectively investigate the incidence of accidental awareness during general anesthesia in selective operation and analyze the influence factors of accidental aware-ness.Methods Patients scheduled for elective surgery requiring general anesthesia with artificial air-way during April 2013-July 2015 were collected.The incidence was obtained from postoperative fol-low-ups.Perioperative data and anesthetic drugs were collected to analyze influence factors. Results A total of 141 294 patients were enrolled.Eighty patients (0.06%)were definitely identified as awareness.Multiple regression analysis showed that awareness during operation was associated with total intravenous anesthesia (OR=5.181,95% CI 3.032-8.853),using laryngeal mask airway (OR=2.478,95% CI 1.544-3.977)and ASA Ⅲ or Ⅳ (OR=9.202,95% CI 5.475-15.466).Pre-medication of midazolam (OR=0.125,95% CI 0.080-0.196)and combination of sevoflurane (OR=0.193,95% CI 0.113-0.330)had lower incidence of awareness during operation.Conclusion Total intravenous anesthesia,using laryngeal mask airway and ASA Ⅲ or Ⅳ are risk factors of accidental awareness during general anesthesia.Premedication of midazolam and combination of sevoflurane may prevent awareness during operation.

Objective To explore the mechanism,clinic features and treatment of type Ⅳ cardiorenal syndrome.Methods The clinical data of one patient with cardiorenal syndrome characterized with chest distress was analyzed.Results After combination treatment,the symptoms were relieved,the amount of physical activity was increased,and the functions of heart and kidney were improved.Conclusions Active,prompt and rational multidisciplinary care can control the progression of cardiorenal syndrome,increase survival rate and improve life quality.

Objective:To investigate the synergistic killer effect of natural killer cells(NK cells) combined with tamoxifen(TAM) on breast cancer cells(BCC)through the experiment in vitro,and to explore its mechanism.Methods:Three kinds of BCC with different receptor expression levels were selected for the experiment.Blank control group, different concentrations of TAM groups and different time groups were set up.MTT assay was used to detect the inhibitory rates of proliferation of cells,and the final experiment concentration of 5 μmol·L-1 was determined.The cells were divided into natural-release group,largest-release group,TAM group,NK cells group, and combined-experimental group(BCC+NK cells+TAM),and the synergistic killer effect of NK cells combined with TAM in different effector-target ratios were detected with Calcein-AM release assay.In ELISA assay the cells were divided into blank control group (NK cells),NK cells+TAM group, NK cells+BCC group and combined-experimental group,and the levels of IFN-γ and TNF-α in the NK cells in various groups were measured.In flow cytometry detection the cells were divided into blank control group (NK cells),NK cells+TAM group,NK cells+ BCC group, and combined-experimental group,and the expression levels of NKp46,CD158a,CD158b,CD158b2,and CD158e were determined;while the cells were divided into blank control group (BCC),BCC+TAM group,BCC + NK cells group, and combined-experimental group,and the expression levels of the MICA,ULBP1 and ULBP2 were detected.Results: The MTT assay results showed that the inhibitory rates of proliferation of 3 kinds of BCC had obvious time-and concentration-dependence (P<0.05).The Calcein-AM release assay results showed that the killing-rates of BCC in TAM groups were increased with the increase effector-target ratios of compared with NK cells group;and the killing-rate in combined experimental group was obviously higher than those in NK cells and TAM groups(P<0.05).The ELISA assay results showed that the levels of TNF-α and IFN-γ of NK cells in various experimental groups with BCC or not were increased compared with blank control group(P<0.05 or P<0.01);the levels were significantly increased when combined with TAM (P<0.05).The flow cytometry results showed that the NKp46 expression levels in various experimental groups were elevated compared with blank control group(P<0.05);the expression levels of CD158a, CD158b,CD158b2, and CD158e were significantly decreased(P<0.05);the expression levels MICA,ULBP1, and ULBP2 in BCC were significantly increased (P<0.05).Conclusion:The NK cells combined with TAM has the synergistic killer effect on the BCC in vitro.The synergetic mechanism may be as follows: TAM could increase the secretion of TNF-α and IFN-γ of NK cells to enhance their cytotoxicity;TAM also could up-regulate the expression levels of activating receptors and activating ligands,and down-regulate the expression levels of inhibitory receptors to increase the killing ability of NK cells.