Objective To discuss a method combining serum‐pharmacology and electrophysiology technology ,and to research the mechanism of dilating porcine coronary artery of sodium tanshinone Ⅱ‐A sulfonate (DS‐201) .Methods To give mice intragastric administration solution and measure DS‐201 concentration in mice serum ,and apply the serum to single channel patch to research its effect on big conductance calcium‐activated potassium channels(BKca ) in porcine coronary artery smooth muscle cells (PCASMCs) . Results The linear range of concentration containing DS‐201 serum was 0 .73 to 1 .91 μg/mL (r=0 .997 7) ,the recycle rate was 99 .85% -101 .47% ,and the concentration was(7 .32 ± 4 .25)μg/mL ;the result indicates that serum containing DS‐201 has activa‐tion effects on BKCa in PCASMCs ,while there was no statistical significance (P>0 .05) .Conclusion The establishment method of the alcohol extraction of Danshen is useful and reliable .The HPLC method of measuring DS‐201 concentration is precise .Choosing higher quality drugs or raising bioavailability can improve combination of the serum pharmacology and electrophysiological tech‐nique .

Objective To research the homology and cross reaction characteristics of human papillomavirus(HPV)16 type L2 N-terminal(1-200)protein in clinical common HPV infection types.Methods The amino acid sequences of the common HPV infection types(6,11,16,18 ,etc.)were blasted and it was found that 1-200 N-terminal sequence of L2 protein was highly homologous.The gene of HPV16 L2(1-200)was amplificated from tissue sample of cervical cancer patient and inserted into the prokaryotic expression vector PGEX-4T-1 to construct the recombinant plasmid PGEX-4T-1-HPV16 L2(1-200).After sequencing identification,the recombinant plasmid was tranformed into E.coli BL21(DE3).Induced by IPTG,the fusion protein containing HPV16 L2(1-200)was expressed and analyzed by both SDS-PAGE and Western blot.Furthermore,the specific binding capacity of the fusion protein to the HPV 6,11,16 and 18 DNA positive patient serums were analyzed by Western blot.The fusion protein was purified with Ni-NTA Agarose Kit and coated with ELISA reaction plates.The specific serum IgG of 98 condyloma acuminatum patients,135 cervix cancer patients and 96 healthy control subjects were detected respectively by indirect ELISA.Results After comparing the amino acid sequences of the common HPV infection types(HPV6,11,16,18,etc.),We found that the homology of HPV L2(1-200)reached 52.7%-74.3%.The recombinant plasmid PGEX-4T-1-HPV 16 L2(1-200)was constructed successfully.Highly expressed HPV16 L2(1-200)fusion protein was obtained and the expression level was account for up to about 22.6% of total bacterial protein.The relative molecular mass(Mr)of the fusion protein is about 49×103,which matches up to the expected Mr Meanwhile,the serums of HPV 6,11,16,18 DNA positive patients were used as the first antibody and the specific band was detected respectively at about 49 × 103 by Western blot.Indirect ELISA showed that the A490 values of the specific IgG of condyloma acuminatum group,cervical cancer group and healthy control subjects were 0.753 ± 0.262,0.756 ± 0.274 and 0.178 ± 0.157 with the positive rate were 89.8%,88.9% and 9.4% respectively.There was no significance of the specific IgG between condyloma acuminatum group and cervical cancer group(P＞0.05),but it was significant among the three groups(P＜0.001).Conclusion The N-terminal 1-200 amino acids of HPV L2 has high homology and there exits cross reaction among the most common HPV infection types.

Objective:To comparatively analyze the efficacy of S-1 plus cisplatin as first-line chemotherapy between advanced AFP posi-tive and AFP negative gastric cancer. Methods:A total of 89 eligible patients with advanced gastric cancer from January 2010 to June 2013 were enrolled in this retrospective study. The cases were divided into AFP positive group (n=18) and AFP negative group (n=71) based on serum AFP level before treatment. Both groups received S-1 plus cisplatin as first-line treatment. Results:Significant differ-ences were observed in remission rate (66.7% versus 32.4%, P=0.028) and radical surgical resection rate (44.5% versus 18.3%, P=0.029) after chemotherapy between AFP positive group and AFP negative group. In the two groups, neither progression-free survival nor overall survival (OS) showed any significance (P>0.05) in patients without surgery after chemotherapy. However, the disease-free survival of the two groups with radical surgical resection after chemotherapy was significantly different (median:27.0 months versus 15.6 months, P=0.034). The OS of the AFP positive group was evidently longer than that of the AFP negative group (median:16.0 months versus 11.0 months, P=0.005). Conclusion:As first-line chemotherapy, S-1 plus cisplatin was more effective for the advanced AFP positive gastric cancer and prolonged overall survival.

Objective To study on FBD (composed with Poria cocos, Atractylodes macrocephala, and Angelica sinensis) used to improve learning and memory function of animals. Methods The mice were treated by ig FBD with the doses of 35, 70 and 140 mg/kg for one week continuously. Effect of FBD on dysmnesia of acquired learning of mice induced by scopolamine, dysmnesia of memory retention of mice induced by NaNO2, and dysmnesia of reappearance of memory of mice induced by 45% ethanol were studied. Improvement for the dysmnesia of memory due to chronic blood deficiency in brain of rats induced by ligating the carotid arteries of two sides were observed. Results FBD can improve the dysmnesia of mice induced by scopolamine, NaNO2, and 45% ethanol significantly, also dysmnesia due to chronic blood deficiency in brain of rats induced by ligating the carotid arteries. Conclusion FBD has the function to improve the learning and memory function of animals.

Objective To investigate the correlation between long non-coding RNA (lncRNA)-LOC391533 and inadequate placental spiral artery remodeling in severe preeclampsia (sPE).Methods Thirty-six gravidas who were admitted to the Third Affiliated Hospital of Zhengzhou University with sPE from January 2016 to December 2016 were enrolled in sPE group.An equal number of healthy gravidas who experienced uneventful pregnancy and were of similar age (difference less than two years) and gestational age (difference less than one week) to those in the sPE group served as controls.Scanning electron microscopy was used to measure the luminal area and vessel wall thickness of placental spiral arteries for all gravidas.Levels of vascular endothelial growth factor (VEGF) and soluble VEGF receptor-1 (sVEGFR-1) in placenta tissues and maternal serum samples were detected by Western blot and ELISA.Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of LOC391533 at mRNA level in placenta tissues of the two groups.Independent two samples t-test,Mann-Whitney U test and Spearman correlation analysis were used for statistical analysis.Results (1) The average luminal area of spiral arteries of the sPE group was smaller than that of the control group [(130.1 22.3) vs (188.1 ±21.5) μ m2,t=10.888,P＜0.05],but the average thickness of spiral artery wall was thicker [(122.619.5) vs (98.9±2.5) μ m,t=-8.812,P＜0.05].(2) Compared with the control group,the sPE group showed increased sVEGFR-1 at protein level in both placenta tissues and serum samples [placenta:0.2±0.0 vs 0.4±0.1,serum:(15.6±2.4) vs (50.8±6.1) ng/L,t=-17.569 and-30.699,both P＜0.05],decreased VEGF at protein level in both placenta tissues and serum samples [placenta:0.6 ± 0.1 vs 0.2±0.0,serum:(40.8±3.2) vs (28.1 ±3.2) ng/L,t=18.013 and 16.200,both P＜0.05],and enhanced expression of LOC391533 at mRNA level in placenta tissues (1.00.2 vs 2.40.5,t=-14.799,P＜0.05).(3) Expression of LOC391533 at mRNA level in placenta tissues of the sPE group was positively correlated with spiral artery wall thickness and levels of sVEGFR-1 protein in placenta tissues and serum (r=0.683,0.759 and 0.857,all P＜0.05),and negatively correlated with luminal area and levels of VEGF protein in placenta tissues and serum (r=-0.702,-0.806 and-0.796,all P＜0.05).Conclusions Abnormal expression of VEGF and sVEGF-1 in placenta and serum of patients with sPE may be related to inadequate placental spiral artery remodeling.

Objective: To explore growth and intelligence development of low birth weight infants (LBWI) at 24 and 36 months of age, so as to provide reference for early monitoring and intervention of the development of LBWI. Methods: A total of 100 LBWI born and managed in Hefei Maternal and Child Health Care Institution were selected from 2012 October 1 to 2015 December 30, and 99 normal birth weight infants (NBWI) under child health management in the same sitinstitution were selected as controls. According a prospective cohort study method, and based on the establishment of a cohort and monitoring of childhood growth and development, a unified method was used to longitudinally follow up and observe the physical fitness of two groups of infants at the determined time points. The development of LBWI and NBWI at 24 and 36 months of age was surveyed using the Gesell Development Scale. Results: Weight, length and head circumference of LBWI children at the age of 15-36 months were significantly lower than those of NBWI children ( P <0.05). In addition, 117 children (43.98%) completed the full assessment of intelligent development scale, including 62 LBWI and 55 NBWI. The scores of Gesell in NBWI group was higher than that in LBWI group at 24 and 36 months of age, including adaptability, gross motor, fine metor skills, language and personal social functions ( t =-4.17, -3.82, -3.21 , -3.03, -2.61; -4.23, -3.16, -3.07, -3.13, -3.99, P <0.05). Multivariate linear regression analysis found that birth weight was positively correlated with adaptability, gross motor, fine motor skills, language functions at 24 and 36 months of age and personal social function at 36 months of age ( β =0.004, 0.010; 0.003, 0.008; 0.003, 0.007; 0.004, 0.009; 0.011, P ＜0.05). Conclusion The growth and development of LBWI children are significantly delayed compared to NBWI children. The scores of LBWI children are lower than those of NBWI children in all functional areas. Weight is the main factor affecting children s intellectual development. Early monitoring and intervention of low birth weight infants should be carried out to avoid or mitigate adverse consequences.

Vitamins are classified as either fat-soluble (vitamins A, D, E, K) or water-soluble (vitamins B and vitamin C). Traditional methods of immunoassay have only been developed for vitamins D,B6, B9 and B12. However, they cannot distinguish between vitamin subtypes such as D2, D3 and associated epi isomers (which has higher leveks in infants),giving false positive or negative results. Mass spectrometry has become a gold standard method for small molecule analysis in biological samples with its advantages in speed,resolution,sensitivity and specificity. It is widely used in clinical research and diagnosis and provides an efficient method for simultaneous detection of multivitamins in one injection using one low volume sample collection.

Objective:To explore the feasibility of applying quantitative flow ratio(QFR) to assess the degree of coronary artery functional stenosis before surgery, and to guide coronary artery bypass grafting(CABG) revascularization strategy.Methods:The study prospectively included a total of 154 patients who were electively treated with CABG in the 11th ward of the Department of Cardiac Surgery of Beijing Anzhen Hospital from January 2019 to September 2020, and their coronary angiography visually showed stenosis of the coronary artery to perform QFR analysis to know the diseased blood vessels. For functional stenosis, the surgeon was blinded to the results of QFR analysis before surgery. Collect its baseline data, perioperative data and recent clinical outcomes for summary analysis.Results:One year later, the coronary artery CTA showed that the occlusion rate of functionally significant disease(QFR<0.8) was 5.5%, and that of non-functionally significant disease(QFR≥0.8) was 15.6%. There was no difference in angina class or repeat interventions between patients with or without occluded bypass grafts.Conclusion:According to QFR analysis, coronary arteries with functional non-significant disease have a higher risk of grafts failure than those with functionally significant disease. For coronary arteries with negative QFR lesions, the risk of occlusion of arterial grafts is higher than that of venous. However, this finding is not significantly related to clinical prognosis, because patients with patency or occlusion of the grafts in non-significant lesions have not found excessive angina pectoris or repeated coronary interventions. QFR-guided selection of coronary surgery strategies is safe and feasible.

Objective To analyze the short-term clinical efficacy and influencing factors of ustekinumab monoclonal antibody(UST)in the treatment of Crohn′s disease(CD).Methods Retrospective cohort study was used to collect the clinical data of CD patients treated with UST in the 10th People′s Hospital affiliated to Tongji University from December 2020 to October 2022.The main analysis is the short-term clinical efficacy and influencing factors of UST treatment for CD at weeks 8 and 16,And analyze the endoscopic response rate of some patients.Results A total of 91 CD patients who first used UST were included.The 8-week clinical response rate of UST treat-ment for CD was 61.5%,and the clinical response rate was 45%;The clinical response rate at 16 weeks was 71.4%,and the clinical response rate was 54.9%.56 cases underwent endoscopic re-examination in our hospital,and the endoscopic response rate at 16 weeks was 41.1%.Univariate analysis showed that fistula(including anal fistula,personal history of anal fistula,and intestinal skin fistula)is associated with clinical remission in Crohn′s disease patients at 8/16 weeks.Further multivariate COX regression analysis showed that the presence of a history of anal fistula surgery was an independent protective factor affecting clinical remission in CD patients treated with UST at 8 weeks(HR = 0.04,95%CI:0.00～0.38;P = 0.005)and 16 weeks(HR = 0.04,95%CI:0.01～0.34;P = 0.003)compared to those without fistula;Narrow lesions are an independent risk factor for 16 week clinical remission in CD patients compared to non-narrow and non-penetrating lesions(HR = 1.75,95%CI:1.08～2.84;P = 0.023).No patients were found to have stopped medication due to serious adverse reactions.Conclusions UST can improve the clinical remission and response of CD patients at 8/16 weeks,and has good short-term clinical efficacy.CD patients with a personal history of anal fistula are recommended to use UST monoclonal antibodies,while patients with stenotic lesions should be cautious in using UST monoclonal antibodies.Whether the patient has undergone surgical treatment in the past,as well as whether UST has been used on the first or non-first line,has no significant impact on clinical remission.

OBJECTIVETo explore the immuno-effect of plasmacytoid dendritic cells (pDC) on bacteria infection induced spontaneous remission (SR) of leukemia.

METHODSBoth pDC and myeloid dendritic cells (mDC) were isolated and purified from leukemic patient with SR and healthy donor by combination of immunomagnetic beads and flow cytometry. pDC were cultured in RPMI1640 medium and stimulated with different bacteria. The T cells proliferation was detected by MTT, and cytokine production by ELISA kits.

RESULTSThe human bacterial pathogen Staphylococcus aureus and Pseudomonas aeruginosa stimulation for 48 h resulted in the maturation of pDC with production of high quantity of IFN-α at (15.34 ± 2.91) ng/ml and (10.38 ± 1.41) ng/ml, respectively, comparing with that of negative group at (1.36 ± 0.13) ng/ml (P<0.01). Activated pDC could promote the differentiation of naive CD4⁺ T cells to Th1 cells with secretion of IFN-γ at (2.16 ± 0.37) ng/ml and (2.73 ± 1.11) ng/ml, respectively, comparing with that of positive control at (2.55 ± 0.23) ng/ml (P > 0.05). Activated pDC showed higher T cell stimulatory capacities [proliferation index (PI) was 4.36 and 4.05, respectively] than that of non-activated pDC (PI was 1.23 and 0.13, respectively) (P < 0.01).

CONCLUSIONStaphylococcus aureus and Pseudomonas aeruginosa activated pDC may play a key role in SR of leukemia following severe infections.

CD4-Positive T-Lymphocytes ; Dendritic Cells ; immunology ; Flow Cytometry ; Humans ; Interferon-alpha ; Leukemia ; diagnosis ; immunology ; Lymphocyte Activation ; Pseudomonas aeruginosa ; immunology ; Remission, Spontaneous ; Staphylococcus aureus ; immunology