OBJECTIVE: To establish a method for determination of the contents of puerarin in infant Fuxieling effervescent granules with HPLC.METHODS:The puerarin was extracted by ultrasonics, using methanol as solvate.The stratographic column was Shim - Pack C1s with acetonitrile - water( 10: 90) as mobile phase. The flow rate was 2.0ml/min and the detecting wavelenghth was 250nm.The temperature of column was 25℃ .RESULTS:The resolution between the puerarin and the adjacent peaks was ＞1.5 .The number of theoretical plates was 18 550 .The standard curve was linear in the range of 0.964μg～7. 712μg .The average recovery was 100.2% (RSD = 1.76%, n = 6).CONCLUSION:The method is simple, rapid and accurate. It can be used for quality control of the preparation.

The inter-species differences of estradiol metabolism were investigated in human, Beagle dog and rat liver microsomes by comparing enzyme kinetics of parent drug and the formation of its major metabolites. The incubation systems of estradiol with liver microsomes of the three species were optimized in terms of estradiol concentration, microsomal protein content and incubation time. The concentrations of estradiol and its metabolites were measured by LC-MS/MS method. The t1/2, CLint, CLh, Km and Vmax of estradiol incubated with male human liver microsomes were 40.02 +/- 8.32 min, 41.39 +/- 6.57 mL x min(-1) x kg(-1), 13.81 +/- 12.36 mL x min(-1) x kg(-1), 26.8 +/- 6.99 micromol x L(-1) and 0.75 +/- 0.92 micromol x L(-1) x min(-1), respectively. The corresponding parameters of female human were 44.71 +/- 10.21 min, 29.85 +/- 8.97 mL x min(-1) x kg(-1), 0.01 +/- 0.68 mL x min(-1) x kg(-1), 44.2 +/- 7.73 micromol x L(-1) and 1.27 +/- 4.41 micromol x L(-1) x min(-1), that of male dog were 21 +/- 7.33 min, 165.53 +/- 29.33 mL x min(-1) xkg(-1), 26.01 +/- 8.39 mL x min(-1) x kg(-1), 19.5 +/- 7.34 micromol x L(-1) and 1.6 +/- 0.65 micromol x L(-1) x min(-1), that of female dog were 25.5 +/- 5.32 min, 135.11 +/- 42.34 mL x min(-1) x kg(-1), 0.24 +/- 3.18 mL x min(-1) x kg(-1), 8.33 +/- 6.32 micromol x L(-1) and 0.51 +/- 2.15 micromol x L(-1) x min(-1), that of male rat were 5.11 +/- 3.84 min, 485.63 +/- 36.52 mL x min(-1) x kg(-1), 49.57 +/- 15.29 mL x min(-1) x kg(-1), 62 +/- 13.74 micromol x L(-1) and 19.16 +/- 9.67 micromol x L(-1) x min(-1), and that of female rat were 7.0 +/- 3.69 min, 354.82 +/- 33.33 mL x min(-1) x kg(-1), 8.04 +/- 3.23 mL x min(-1) x kg(-1), 35.38 +/- 7.65 micromol x L(-1) and 8.39 +/- 4.91 micromol x L(-1) min(-1), respectively. There were nine metabolites detected from all the three species, but the relative amounts of the metabolites generated were different in three species. The results indicted that the major phase I metabolic pathway of estradiol was similar in the liver microsomes from all the three species. However, the inter-species differences were found in the view of relative amounts of the metabolites as well as the metabolic characteristics of estradiol in liver microsomal incubation.

Objectives To investigate the effects of different degrees of carotid artery stenosis on cognitive function and neuronal apoptosis of hippocampal CA1 region in rats and to analyze the possible mechanisms of cognitive impairment. Methods According to the random number table,50 male Wistar rats were allocated into a sham operation group,a unilateral mild stenosis group,a unilateral moderate stenosis group,a unilateral severe stenosis group,a bilateral mild stenosis group,a bilateral moderate stenosis group,
a bilateral severe stenosis group,and a sham operation group. Aneedle-controlled suture method was used to induce a carotid stenosis model with different degrees of stenosis in rats. Water maze was to localize navigation and spatial search test was used to evaluate the cognitive function with different degrees of carotid artery stenosis in rats. Immunohistochemical method was used to observe the numbers of positive cells of P75 neurotrophin receptor (p75NTR),Bax,Bcl-2,neurotrophic factor-3 (NT-3 ),nerve growth factor (NGF)in hippocampal CA1 region under the light microscope. The conditions of neuronal apoptosis were observed. Results In 50 rats,6 died,1 rat in poor health failed to complete the Morris water maze test. The degree of bilateral carotid artery stenosis in 1 rat failed to meet 30%-60% at the same time,they were all removed. The remaining 42 rats were 6 in each group. (1)Compared with the sham operation group,the mean escape latency was prolonged (39 ± 6 s,32 ± 5 s,69 ± 7 s,respectively vs. 23 ± 4 s;all P < 0. 01),and the percentage of swimming distance in the platform quadrant was decreased (35 ± 4%,44 ± 4%,22 ± 5%,respectively vs. 53 ± 7%;all P < 0. 01),and the cognitive function was decreased with the degree of stenosis in the unilateral severe stenosis group,bilateral moderate stenosis group,and bilateral severe stenosis group. Compared with the unilateral severe stenosis group,the mean escape latency was prolonged and the percentage of swimming distance in the platform quadrant was decreased in the bilateral severe stenosis group (P < 0. 01). (2)Compared with the numbers of positive cells of Bax,p75NTR and Bcl-2 (8. 8 ± 3. 1,4. 2 ± 2. 3,and 5. 8 ± 1. 8,respectively)in the sham operation group,the numbers of positive cells of Bax,p75NTR,and Bcl-2 were increased (25. 5 ± 3. 5,11. 0 ± 2. 2,12. 3 ± 2. 7;15. 8 ± 3. 7,8. 9 ± 2. 2, 10. 5 ± 2. 9;and 47. 9 ± 6. 3,24. 7 ± 3. 0,12. 8 ± 2. 5,respectively)in the unilateral severe stenosis group, bilateral moderate stenosis group,and bilateral severe stenosis group (all P < 0. 01). The numbers of Bax and p75NTR positive cells were increased with the degree of stenosis. When the stenosis was severe,the numbers of Bax and p75NTR positive cells were increased in the bilateral severe stenosis group compared with those of the unilateral severe stenosis group (P < 0. 01). The numbers of NGF and NT-3 positive cells in each stenosis group had an increased trend compared with sham operation group,but there were no significant differences (F =1. 034,and 1. 358;P = 0. 420 and 0. 259 respectively). Conclusions Carotid stenosis can cause cognitive disorder in rats,and it is correlated with the degree of carotid stenosis. Ischemia caused neuronal apoptosis in hippocampal CA1 region may be one of the mechanisms of cognitive impairment after carotid artery stenosis in rats.

Objective To compare the effects of intensive blood pressure (BP) lowering and guideline-recommended standard BP lowering on the early reperfusion and prognosis after intravenous thrombolysis in patients with acute ischemic stroke. Methods This is a randomised controlled trial consisting of 118 consecutive patients who came from Department of Neurology, Nanjing First Hospital from July 2012 to April 2016, accepting intravenous recombinant tissue plasminogen activator thrombolysis with the systolic blood pressure (SBP) being 150-185 mmHg(1 mmHg=0.133 kPa). The patients with ischemic stroke were diagnosed by multi-mode MRI and confirmed to have ischemic penumbra. The SBP of patients randomly assigned to intensive BP lowering group and guideline BP lowering group was maintained in 140-150 mmHg or below 180 mmHg respectively for 72 h and all patients needed to reexamine multi-mode MRI at 24 h. The primary endpoints were the neurologic function at early stage, modified Rankin Scale (mRS) score and the mortality at 90 d;the secondary endpoints were the volume of infarction and hypoperfusion area, the rate of reperfusion, hemorrhagic transformation (HT) and syptomatic intracerebral hemorrhage (sICH). Results Forty-nine cases in intensive BP lowering group and 56 cases in guideline BP lowering group acquired the available images. The volume of infarction was increased both in these two groups, and there was no statistically significant difference in the increased values ((13.21±9.51) cm3 vs (12.95±9.68) cm3). There were no statistically significant differences in the volume of hypoperfusion, reperfusion rate, neurologic function at early stage, the mRS scores and mortality at 90 d, the incidence of sICH except the rate of HT (9.4%, 5/53 vs 23.1%, 15/65, χ2=3.860, P=0.049) between the two groups.Conclusion Early intensive BP-lowering treatment has no adverse effects on the transformation of ischemic penumbra and prognosis after intravenous thrombolysis in patients with acute ischemic stroke and may decrease the the rate of HT in some degree.

Objective To observe the effects of Xifeng Tongnao capsules on the content of TNF-α and IL-1β in brain tissue of rats with focal cerebral ischemia-reperfusion injury. Methods A total of 120 adult SD rats were randomized into 6 groups: Xifeng Tongnao low-, middle- and high-dose groups, model control group, Buchang Naoxintong group and sham-operated group.Buchang Naoxintong group were treated with Buchang Naoxintong capsules at 0.864 g??kg-1.Xifeng Tongnao high-, middle-and low-dose groups were treated with 3.456, 1.728, and 0.864 g??kg-1 Xifeng Tongnao capsules, respectively;sham-operated group and model control group were treated with equal volume of purified water. Medications were administered intragastrically once daily for 7 days. The acute transient focal cerebral ischemia-reperfusion model was established by middle cerebral artery occlusion ( MCAO) 1 h after the final dose, and rats in the sham-operated group only received anesthesia and stripping without occlusion.All rats were sacrificed after reperfusion for 24 h, and expression levels of TNF-α and IL-1β in the brain tissue were detected by ELISA. Results TNF-αcontent in Xifeng Tongnao capsutes low-, middle-and high-dose groups were (35.34±8.95), (33.75±6.92), and (40.95±5.39) ng??L-1, respectively.IL-1β content were (1.44±0.47), (1.45± 0.23), and (1.61±0.33) ng??L-1 in low-, middle- and high-dose groups, respectively.TNF-α and IL-1β were (38.96±9.84) and (1.56±0.31) ng??L-1, respectively in Buchang Naoxintong group, (52.74±6.76) and (2.79±0.45) ng??L-1in the model control group, and (32.54±4.00) and (1.32±0.22) ng??L-1 in sham-operated group.TNF-αand IL-1βcontent were significantly higher in Buchang Naoxintong group than in sham-operated group ( P<0. 05 ) . TNF-α and IL-1β content were significantly decreased in Xifeng Tongnao high-, middle- and low-dose groups (all P<0.05). Conclusion Xifeng Tongnao capsules can protect brain tissue by supressing TNF-α and IL-1β and alleviating inflammatory injury from ischemia.

The association between serum fetuin A and diabetic nephropathy (DN) in patients with type 2 diabetes mellitus(T2DM) was investigated.Serum levels of fetuin-A and adiponectin were measured by ELISA and the clinical data were collected in 101 T2DM patients with normal albuminuria (NA group),87 patients with microalbuminuria (MA group),44 patients with overt albuminuria (OA group),and 130 normal control subjects (NC group).Fetuin-A levels increased progressively from NC,NA,MA,to OA groups,and there were significant differences among groups(P＜0.05 or P＜0.01).Fetuin-A level in patients was positively associated with duration of illness,urinary albumin to creatinine ratio (ACR),fasting blood glucose,2-hour postprandial blood glucose,and C-reactive protein(CRP,P＜0.05 or P＜0.01),but was negatively associated with age and adiponectin (all P＜ 0.01).Multiple stepwise regression analysis indicated that duration of diabetes,fetuin-A,CRP,and adiponectin were independent determinants for ACR (all P ＜ 0.01).Fetuin-A may be involved in the development and pathogenesis of DN by proinflammatory effects and repressing adiponectin production.

Tumor metastasis is a primary cause of death among solid tumor patients.Interventions such as chemotherapy with cytotoxic drugs and other targeted therapies focusing on specific genes or tumorigenesis can induce tumor shrinkage in most cases.However,these interventions do not substantially prolong the lives of patients.In recent years,immunotherapy has made significant breakthroughs in treating solid tumors with the introduction of immune checkpoint inhibitors(ICIs),following the clinical use of Ipilimumab,the first anti-tumor drug with ICIs initially approved in 2011.Currently,four types of ICIs including CTLA-4,PD-1,PD-L1,and LAG3 have been clinically applied and have demonstrated specificity towards more than 20 types of solid tumors.The significance of ICIs lies in their superiority to conventional drugs in three aspects:(1)They can prolong the survival of late-stage patients and even achieve"clinical cure";(2)They allow patients with rare tumors access a pan-tumor treatment based on specific biomarkers;and(3)They can free a majority of patients from invasive surgical approaches to preserve organ functions,thereby improving patient's quality of life.This article provides a summary of multiple clinical research projects and explores the clinical evidence derived from the notable achievements related to the three advantages.

Objective:To investigate the correlation between the door-to-needle time (DNT) delay and the short-term functional outcome after intravenous thrombolysis in patients with minor ischemic stroke and the influencing factors of DNT delay.Methods:From October 2016 to May 2018, patients with minor ischemic stroke received intravenous thrombolysis with alteplase from the Stroke Database of Nanjing First Hospital were enrolled retrospectively. DNT delay was defined as DNT > median. The modified Rankin Scale was used to evaluate the short-term functional outcome at 3 months after stroke. 0-1 was defined as good outcome, and ≥2 was defined as poor outcome. Univariate analysis was used to evaluate the correlation between DNT delay and short-term functional outcome. Multivariate logistic regression analysis was used to evaluate the possible influencing factors of DNT delay. Results:A total of 102 patients with minor ischemic stroke were enrolled. The median DNT was 40 min, 36 patients (35.3%) had DNT delay, and 27 patients (26.5%) had poor short-term outcome. Univariate analysis showed that there was no significant difference in the proportion of patients with DNT delay between the poor outcome group and the good outcome group (44.4% vs. 32.0%; χ2=1.346, P=0.252). Multivariate logistic regression analysis showed that there was a significant independent negative correlation between hypertension and DNT delay (odds ratio 0.359, 95% confidence interval 0.137-0.939; P=0.037). Conclusion:For patients with minor ischemic stroke receiving intravenous thrombolysis, DNT delay is not associated with the outcome. The absence of hypertension may be one of the factors affecting the DNT delay.

Objective To investigate the diagnostic value of detection of protein SP70 in differentiating benign and malignant pleural effusion.Methods A case-control study was conducted from July 2011 to February 2012.108 cases of pleural effusion from patients with clinically proven lung cancers and 122 cases of benign pleural effusion were collected.SP70 was detected by Sandwich ELISA,while CEA,CYFRA21-1,NSE were measured by electrochemiluminescence immunoassay for comparison.Meanwhile,protein SP70 on exfoliated cells in pleural effusion was detected by direct immunofluorescence,and was compared with the results of HE staining.The differences between the groups were evaluated by the chisquare test Fisher' s exact test.Results Positive rates of SP70,CEA,CYFRA21-1,NSE were 72.2％,58.3％,52.8％ and 30.6％ in malignant pleural effusion,obviously higher than benign pleural effusio (9.8％,13.1％,23.0％ and 19.7％).The specificity of SP70,CEA,CYFRA21-1,NSE were 90.2％,86.9％,77.0％ and 80.3％,NSCLC had significantly higher positive rate than SCLC(74.3％ ＞0.0％,P =0.02 ＜ 0.05),detection of protein SP70 in malignant pleural effusion had significantly higher coincidence rate than HE staining(72.2％ vs 47.2％,x2 =14.03,P ＜ 0.05).Conclusion Determination of the protein SP70 in pleural effusion and in exfoliated cells,can improve the sensitivity and specificity of the diagnosis of malignant pleural effusion.