Objective: To explore change of hemoglobin (Hb) concentration, the relationship between anemia and left ventricular function and the influence of anemia on prognosis in patients with diastolic heart failure (DHF). Methods: According to NYHA classification, a total of 176 DHF patients were divided into class Ⅱ group (n=78), class Ⅲ group (n=50) and class Ⅳ group (n=48), then Hb level and morbidity rate of anemia were analyzed in each group. According to diagnostic standard of anemia, patients were divided into anemia group (n=58, , occupied 33.0%) and non anemia group (n=118, occupied 67.0% ). Left ventricular diastolic function, mortality rate and rehospitalization rate during follow-up were compared and analyzed between two groups. Results: Along with cardiac function class rose (from class Ⅱ to class Ⅳ), Hb level showed a decreasing trend [(130±6) g/L vs. (108±4) g/L vs. (96±12) g/L], while morbidity rate of anemia gradually rose (8.97% vs. 36.0% vs. 68.8%), P<0.05 all in anemia group；Compared with non-anemia group, there were significant rise in percentages of patients with coronary heart disease (55.1% vs. 65.5%), levels of creatinine [(87.6±39.2) μmol/L vs. (113.7±59.8) μmol/L] and N terminal pro B type natriuretic peptide [NT-proBNP, (578.0±136.7) pg/ml vs. (886.0±174.8) pg/ml], and early-diastolic peak velocity deceleration time [(137±15)ms vs. (196±13)ms], and significant reduction in mitral early/late diastolic peak flow velocity [E/A, (0.87±0.32) vs. (0.62±0.29)], P<0.05 all. Compared with non-anemia group, there were significant rise in mortality rate (9.3% vs. 20.7%) and rehospitalization rate (18.6% vs. 32.8%) in anemia group during follow-up, P<0.05 all. Conclusion: DHF patients often complicate with anemia. Along with heart failure aggravates, their morbidity rate of anemia rises, and anemia may aggravate cardiac diastolic dysfunction. Mortality rate and rehospitalization rate rise in DHF patients complicated with anemia.

Objective To screen B and T cell antigen epitopes on Acinetobacter baumannii outer membrane protein 33×103-36×103 (OMP33-36).Methods B and T cell epitopes on OMP33-36 of Acinetobacter baumannii were predicted by bioinformatics methods and synthesized.Recombinant expression plasmid pET-30a-OMP33-36 was cloned and used to express OMP33-36 in a prokaryotic expression system.The expressed OMP33-36 was used to immunize BALB/c mice after purification.Serum sample was collected from each mouse in immunization and negative control groups, and then analyzed by indirect ELISA with synthesized peptides to identify B cell epitopes.Splenocytes were separated from every mouse and then cultured with each of the synthesized peptides, respectively.Double sandwich ELISA was performed to detect IFN-γ secretion in the supernatant of cell cultures for screening of T cell epitopes.Results Candidates of B and T cell epitopes were constructed, which were PB1, PB2, PB3, PT1, PT2 and PT3.Results of the indirect ELISA showed that peptides PB1 and PB2 reacted with the serum samples collected from immunized mice and A450 values of the immunization group were significant higher than those of the negative control group.Compared with the negative control group, enhanced secretion of IFN-γ following peptide PT3 stimulation was observed in the immunization group as indicated by the double sandwich ELISA.Conclusion Two B cell epitopes PB1 and PB2, and one T cell epitope PT3 on the OMP33-36 of Acinetobacter baumannii were successfully constructed and screened out.

Objective: To investigate whether Pseudomonas aeruginosa recombinant protein PA3611 can induce the in vitro epithelial-mesenchymal transformation (EMT) of rat bronchial epithelial cells. Methods: A series of processes including DNA synthesis, gene amplification, vector construction, induction of expression and protein purification was used to synthesize the recombinant protein PA3611. CCK-8 kit was used to detect the proliferation of bronchial epithelial cells after stimulation with the protein PA3611. Morphological changes in bronchial epithelial cells were observed. Western blot and qPCR were performed to measure the expression of E-cadherin (E-CAD) and α-smooth muscle actin (α-SMA). T test was used for statistical analysis. Results: DNA sequencing verified the successful preparation of the recombinant protein PA3611. PA3611 inhibited the proliferation (P<0.05) and induced the EMT of bronchial epithelial cells. Moreover, it also inhibited the expression of E-CAD protein, but promoted the expression of α-SMA (P<0.05). Conclusion The recombinant protein PA3611 can induce the EMT of bronchial epithelial cells and the underlying mechanisms are worthy of further investigation.

OBJECTIVETo study the stability of ginsenoside Rg1 before and after being modified by PEG (PEG-Rg1) in isolated rat stomachs.

METHODSD rats, after 18 h of fasting, were randomly divided into the Rg1 group and the PEG-Rg1 group. Rg1 stomach perfusion fluid and PEG-Rg1 infusion fluid were accurately extracted and injected into their stomachs in vitro, with oscillation at 37 degrees C. Samples were taken in different time points and contents of ginsenoside Rg1 were determined by UPLC to observe and compare the stability of ginsenoside Rg1 and PEG-Rg1 in rat stomachs in vitro.

RESULTRg1 in rat stomachs showed poor stability, Rg1 was measured to be 26.8% in 2 h, with degradation of 73.2%. Its stability in PEG-Rg1 was improved in rat stomachs, Rg1 was measured to be 81.8% in 2 h, with degradation of only 18.2%.

CONCLUSIONPEG-modified ginsenoside Rg1 can enhance the stability of ginsenoside Rg1 in stomach and improve degradation and poor stability of ginsenoside Rg1 in stomach.

OBJECTIVETo investigate targeted distribution of ginsenoside Rg1 in mice tissues before and after modification by the PEG.

METHODSD mice were randomly divided into two groups and given Rg1 and PEG-Rg1 by intravenous injection respectively. Their samples of blood and organ tissues were taken at different time points. The content of Rg1 in samples were determined by UPLC and used as indicator to observe the targeted distribution of Rg1 in mice tissues.

RESULTThe AUC of ginsenoside Rg1 in tissues of the Rg1 group were in the order of liver, kidney, lung, heart and spleen, with the liver targeting coefficient was of 2.01. While the AUC of ginsenoside Rg1 in tissues of the PEG-modified group were in the order of the kidney, liver, lung, heart, spleen, with the liver targeting coefficient was of 9.21.

CONCLUSIONPEG modified Rg1 can increase Rg1's targeting selectivity to the liver, kidney and lung in mice.

Objective To investigate the Rv2346c gene function through constructing Rv2346c gene knockout strains of Mycobacterium tuberculosis (M . tuberculosis) mediated by bacteriophage and observing its virulence after infecting mice lung tissue in vivo .Methods The affinal exchange sites (AES) of the target gene was built ,and then integrated into the phage genomes of M .tuberculosis for harvesting the phagemids .The phagemids was imparted into Mycobacterium smegmatis to get recombinant phages with the same AES .A high titer of the recombinant phages was harvested through amplification in vitro . The M .tuberculosis was transfected and coated on solid medium with hygromycin resistance and cultured for 4 weeks at 37℃ .Single clone was picked out and gene knock-out was confirmed by PCR . Then C57BL/6J mice were infected with either wild type strain (WT ) or knockout strain (KO ) of M . tuberculosis .Mice mortality ,lung tissue inflammation and colony-forming units (CFU ) counts in vitro were observed 6 to 8weeks post infection with different strains . Paired-samples t test was used for comparison between groups ,chi-square test was used for comparison of rates .Results The products of PCR and inserted fragment sizes were consisted with the expectation and confirmed to be the target gene . The target fragment of Rv2346c was removed successfully and the mice were infected for 6-8 weeks .Themice infected with Rv2346c KO strain had reduced mortality (53％ vs 20％ ,χ2 =6 .1112 ,P<0 .05) ,lung tissue inflammation (1040 ± 89 vs 1960 ± 56 ,t=7 .1016 ,P<0 .05) and CFU count in vitro (15 .0 ± 0 .8 vs 90 .0 ± 1 .5 ,t=23 .0361 , P<0 .05) compared with WT strain 6-8 weeks post infection .Conclusion Rv2346c gene knockout strains of M . tuberculosis mediated by bacteriophageis are successfully constructed ,which establishes the foundation for the future gene function study of Rv2346c .

A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of thirteen components including three stilbenes (stilbeneglucoside, polydatin, resveratrol), four anthraquinones (emodin, physcion, emodin-8-β-D-glucopyranoside, aloe-emodin), five flavonoids (epicatechin, rutin, hyperoside, astragalin,quercetin) and one phenolic acid (gallic acid) in Polygoni Multifori Caulis.The separation was carried out on a Waters BEH C18 column（2.1 mm×100 mm,1.7 μm）with gradient elution of acetonitrile-water (0.1% acetic acid) at a flow rate of 0.25 mL•min⁻¹, and column temperature was 35 ℃. The target compounds were analyzed by multiple reaction monitoring (MRM) mode. TOPSIS analysis ware performed to evaluate the samples from different areas and commercial herbs according to the contents of thirteen components. The correlation coefficients of all the calibration curves were higher than 0.991 5. The average recoveries ranged from 95.24% to 102.3%, and the relative standard deviations were less than 5%. The result of TOPSIS analysis showed that the comprehensive quality of Polygoni Multifori Caulis sample from Guangzhou was better. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances, which provided a new basis for the comprehensive assessment and overall control of the quality of Polygoni Multifori Caulis.

[ Abstract]Automatic and accurate segmentation of lung parenchyma is essential for assisted diagnosis of lung cancer. In recent years, researchers in the field of deep learning have proposed a number of improved lung parenchyma segmentation methods based on U-Net. However, the existing segmentation methods ignore the complementary fusion of semantic information in the feature map between different layers and fail to distinguish the importance of different spaces and channels in the feature map. To solve this problem, this paper proposes the double scale parallel attention (DSPA) network (DSPA-Net) architecture, and introduces the DSPA module and the atrous spatial pyramid pooling (ASPP) module in the "encoder-decoder" structure. Among them, the DSPA module aggregates the semantic information of feature maps of different levels while obtaining accurate space and channel information of feature map with the help of cooperative attention (CA). The ASPP module uses multiple parallel convolution kernels with different void rates to obtain feature maps containing multi-scale information under different receptive fields. The two modules address multi-scale information processing in feature maps of different levels and in feature maps of the same level, respectively. We conducted experimental verification on the Kaggle competition dataset. The experimental results prove that the network architecture has obvious advantages compared with the current mainstream segmentation network. The values of dice similarity coefficient (DSC) and intersection on union (IoU) reached 0.972 ± 0.002 and 0.945 ± 0.004, respectively. This paper achieves automatic and accurate segmentation of lung parenchyma and provides a reference for the application of attentional mechanisms and multi-scale information in the field of lung parenchyma segmentation.
Algorithms ; Image Processing, Computer-Assisted/methods* ; Lung/diagnostic imaging* ; Neural Networks, Computer ; Tomography, X-Ray Computed/methods*

Objective:To evaluate the clinical efficacy and safety of Longmu Zhuanggu granule for the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency. Method:This multicenter stratified， block-randomized， double-blind， double-dummy， positive drug （pidotimod granule） parallel controlled， and non-inferiority trail intended to included 240 children patients and divided them into the experimental group （n=120） and the control group （n=120） at the ratio of 1∶1. Patients in both groups were treated for eight successive weeks and followed up for 12 months. The cure rates， numbers of respiratory infections， average courses of disease， curative effects of traditional Chinese medicine （TCM） syndrome， curative effects of individual symptoms， curative effects of immune indexes， and safety indexes between the two groups were observed and compared. Result:A total of 237 subjects were collected from 10 research centers， including 119 cases in the control group and 118 in the experimental group. There were 236 cases enrolled into the full analysis set （FAS）， 210 into the per-protocol set （PPS）， and 236 into the safety set （SS）. The baseline data of the two groups were not significantly different from each other， indicating that they were comparable. The cure rates of the experimental group and control group were 75.21% （88/117） and 73.95%（88/119）， respectively， with the 95% confidence interval （CI） of difference between the two groups being 1.26% （-9.85%，12.37%） for FAS and 3.81% （-6.28%，13.90%） for PPS. The 95% CI fell within the 10% non-inferiority margin， implying that non-infertility test of the cure rate in the treatment of endpoint disease was valid， and the conclusions of FAS and PPS analysis were consistent. There was no significant difference in the number or course of upper respiratory infection， bronchitis， and pneumonia. The difference in curative effects of TCM syndrome between the two groups after four weeks of treatment was not remarkable. After eight weeks of treatment， the total effective rate of the experimental group was 84.62%（99/117）， statistically higher than 78.15%（93/119） of the control group（χ²=-3.26，P<0.05）. There were no significant differences in the disappearance rates of individual symptoms between the two groups after four weeks of treatment. After eight weeks of treatment， the experimental group and control group exhibited the disappearance rates of 67.50%（54/80） and 47.37%（36/76） for shortness of breath and laziness to speak， 75.00%（54/72） and 53.33%（40/75） for poor appetite， 54.55%（60/110） and 37.84%（42/111） for hyperhidrosis， respectively， with obviously better outcomes observed in the experimental group （P<0.05，P<0.01）. The inter-group comparison revealed significant differences in immune indexes after eight weeks of treatment. As demonstrated by comparison with the situations before treatment， IgA， IgG， IgM， and CD4 did not change significantly after treatment. Except for CD8 in the experimental group (P<0.05), there was no significant difference in other immune indexes before and after treatment There was no significant difference in the incidence of adverse reactions. Conclusion:Longmu Zhuanggu granule is not inferior to pidomod granule in the treatment of children recurrent respiratory infection due to lung-spleen Qi deficiency， and it exhibits good safety， implying its promising clinical application value.

Brain tumor incidence shows an upward trend in recent years; brain tumors account for 5% of adult tumors, while in children, this figure has increased to 70%. Moreover, 20%-30% of malignant tumors will eventually metastasize into the brain. Both benign and malignant tumors can cause an increase in intracranial pressure and brain tissue compression, leading to central nervous system (CNS) damage which endangers the patients' lives. Despite the many approaches to treating brain tumors and the progress that has been made, only modest gains in survival time of brain tumor patients have been achieved. At present, chemotherapy is the treatment of choice for many cancers, but the special structure of the blood-brain barrier (BBB) limits most chemotherapeutic agents from passing through the BBB and penetrating into tumors in the brain. The BBB microenvironment contains numerous cell types, including endothelial cells, astrocytes, peripheral cells and microglia, and extracellular matrix (ECM). Many chemical components of natural products are reported to regulate the BBB microenvironment near brain tumors and assist in their treatment. This review focuses on the composition and function of the BBB microenvironment under both physiological and pathological conditions, and the current research progress in regulating the BBB microenvironment by natural products to promote the treatment of brain tumors.