This article was aimed to study the impact on substance and energy metabolism by chemical split fractions of Mori Cortex among hypoglycemic diabetic mouse model, in order to explain the new hypothesis of the science connotation in nature and flavor of traditional Chinese medicine (TCM). Male Kunming mice were intraperitoneally injected with a large dose of streptozotocin (STZ) (170 mg·kg-1) to establish type 1 diabetes mellitus mouse model. Medication was given consecutively for four weeks. The enzyme-linked immunesorbent assay (ELISA) was used to detect glucosekinase (GCK), glycogen phosphorylase (PYGL), pyruvate dehydrogenase (PDH), α-ketoglutarate dehydrogenase (α-KGDHC), phosphoglycerate kinase (PGK), acetyl coenzyme A (acetyl-CoA), adenylate kinase (ADK), fumarase (FUM), cytochrome C reductase (CCR), cytochrome C oxidase (COX) and other indicators. Enzymatic detection was used to determine the content of ATP coenzyme (ATPs), the content and ratio of NAD and NADH, the content of myocardial cell Na+-K+ ATP enzyme, as well as the content of ATP and ADP. The results showed that in the model group, the expression of PYGL was increased; and the expressions of GCK and PDH were decreased. It prompted that the source of glucose increased and the expelling of glucose decreased. The glucose level was increased. The COX expression was reduced and the respiratory chain was blocked. It regulated oxidative phosphorylation and the substrate phosphorylation level. It upregulated the expression of CCR, ATPs, NAD+, PGK, α-KGDHC and ADK. However, the expression of FUM was decreased. The activity of Na+-K+ ATPase was decreased significantly. At last, the metabolic disorders appeared. Mori Cortex aqueous extracts and the chemical split fractions significantly increased the GCK and PDH level in substance metabolism among diabetic mice. The levels of PYGL, α-KGDHC, PGK and acetyl-CoA were decreased (P < 0.05, or P <0.01). Meanwhile, it increased ATP and FUM, myocardial cell Na+-K+ ATP enzyme, and COX level in the energy metabolism (P < 0.05). It decreased the level of NAD+, CCR and ATPs (P < 0.05, or P <0.01). It was concluded that both the aqueous extracts and chemical split fractions of Mori Cortex can effectively improve the substance and energy metabolism disorders of diabetic mouse model. This effect may be related to the cold nature of Mori Cortex.

Objective:To detect the expression of tumor necrosis factor alpha in (TNF alpha) breast cacner and its relationship with imaging features.Methods:Using immunofluorescence and flow cytometry to detect the expression of TNF-α in MDA-MB-231 and MCF-7.Collect 82 patients with mammary gland disease,which was confirmed by pathological tissue,its pathological data,imaging data,and by immunohistochemistry to detect the expression of TNF-α in breast tissues,and analyze and the relationship between its expression and the pathological features and imaging characteristics.Results:TNF-α high expression in MDA-MB-231,the expression of TNF-oα in malignant breast tumor tissue significantly higher than that in benign tumor,the expression quantity associated with lymph node metastasis,TNM stages,strengthen uniform in MRI,the boundary and the shape of the X-ray Mammography (P=0.01),and color flow signal strength in ultrasound (P＜0.05).Conclusions:TNF alpha in breast tumor tissue was unusually high expression,and is closely related to some of the imaging features of breast tumor.

Objective To identify the risk factors of Burkholderia cenocepacia associated nosocomial lower respiratory tract infections (NLRTIs),and to investigate the drug resistance of Burkholderia cenocepacia strains.Methods A total of 138 patients with Burkholderia cenocepacia associated NLRTIs and 40 patients with non-Burkholderia cenocepacia associated NLRTIs were enrolled in the study.All patients were collected from the Second Affiliated Hospital of Wenzhou Medical University during January 2009 and December 2012.Clinical data and results of drug sensitivity tests were retrospectively reviewed.Chi-square test and Logistic regression analysis were performed to identify the risk factors of Burkholderia cenocepacia associated NLRTIs.Results Logistic regression analvsis showed that combination use of 2 or more antimicrobial agents,mechanical ventilation,stay in intensive care unit (ICU) for more than two weeks,use of antacid H2 antagonist and deep venous puncture were the independent risk factors of Burkholderia cenocepacia associated NLRTIs (OR =6.315,5.957,5.254,4.585 and 2.017,P ＜0.05).Burkholderia cenocepacia strains were sensitive to levofloxacin,ceftazidime and sulfamethoxazole; More than 40％ strains were resistant to cefotaxime,ceftriaxone,cefepime,aztreonam and tetracycline; And nearly 100％ strains were resistant to gentamicin,amikacin and tobramycin.Conclusion Burkholderia cenocepacia associated NLRTIs are more likely to occur in patients with combination use of 2 or more antimicrobial agents,mechanical ventilation,and those who stay in ICU for more than two weeks,or received antacid and deep venous punctures,and most Burkholderia cenocepacia strains are multiple drug resistant.

Objective:To assess the left ventricular (LV) systolic function in amateur marathoners by two-dimensional speckle tracking echocardiography.Methods:A total of 59 amateur marathon runners were recruited from January 2019 to June 2020 in Hangzhou and were divided into group A (>2-5 years) and group B (>5 years) based on their time of participating in marathon. Thirty-one healthy volunteers were enrolled from Affiliate Hospital of Hangzhou Normal University during the same period as a control group. Conventional echocardiography combined with two-dimensional speckle tracking imaging were applied to all the subjects to obtain interventricular septum diastolic thickness (IVSd), LV posterior wall thickness(PWd), LV end-diastolic diameter (LVEDd), relative wall thickness(RWT) and LV mass(LVM), LV end-diastolic volume(EDV), LV end-systolic volume(ESV) and stroke volume(SV), LV ejection fraction(LVEF), LV global longitudinal strain (GLS), and global circumferential strain (GCS). Pre-marathon, 1 hour and 4th day post-marathon echocardiography were performed in amateur marathon runners.Results:Compared with the control group, group A amateur marathon runners showed significant increases in IVSd, PWd, LVEDd, RWT and LVM(all P<0.05); In addition, the IVSd, PWd, LVEDd, RWT, LVM, EDV, ESV and SV in group B runners were further increased compared to those of group A runners (all P<0.01). The LVEF and pre-marathon GCS showed no statistically significant differences among the three groups (all P>0.05), while the pre-marathon GLS showed a statistically significant difference among the three groups ( P<0.01). Compared with group A runners, the pre-marathon, 1 hour and 4th day post-marathon GLS and 1 hour post-marathon GCS were significantly decreased in group B runners (all P<0.01); In intra-group comparison, 1 hour post-marathon GLS was significantly decreased in relative to pre-marathon and 4th day post-marathon GLS in both group A and B (all P<0.05). One hour post-marathon GCS was significantly decreased compared with pre-marathon GCS in group B ( P<0.05). Conclusions:Amateur runners who have participated in long time marathon have reduced LV longitudinal strains and transient post-marathon decreases in LV systolic function.

Objective:To investigate the value of radiomics based on unenhanced CT texture analysis in predicting the WHO/International Society of Urological Pathology (ISUP) grading of clear cell renal cell carcinoma (ccRCC).Methods:Postoperative pathology-confirmed ccRCC subjects ( n=90) who received CT scanning and had a definite pathological grading in Cancer Hospital of the University of Chinese Academy of Sciences were collected retrospectively from December 2016 to May 2019. The cases were randomly divided into training group ( n=63) and test group ( n=27) as a ratio of 7∶3. All cases were classified into low grade (grades Ⅰ and Ⅱ, n=57) and high grade (grades Ⅲ and Ⅳ, n=37) according to the new pathological grading (WHO/ISUP grading, version 2016) of renal carcinoma. 3D-ROI segmentation was performed on unenhanced CT images and 93 texture features were extracted. The least absolute shrinkage and selection operator (LASSO) regression was used to reduct dimension of texture parameters and then the radiomics score (Rad-score) was established. The logistic regression was used to develop the prediction model with the pathological grading as the gold standard. The ROC curve and calibration curve were used to evaluate the predictive performance of the model, and the area under the curve (AUC), accuracy, sensitivity and specificity were calculated. The Hosmer-Lemeshow test was used to evaluate calibration degree of the model. Results:The 10 non-zero coefficient texture features were screened out through dimension reduction steps. The Rad-score was formed according to the linear combination of these ten features and corresponding coefficients, and then the prediction model was developed. The AUC of the model in training group was 0.933 (95%CI 0.862-1.000), the sensitivity was 92.3%, the specificity was 89.2%, and the model accuracy was 90.5%. The calibration curve showed the good calibration ( P=0.257). The AUC value in test group was 0.875 (95%CI 0.734-1.000), the sensitivity, specificity and accuracy were 72.7%, 87.5% and 81.5%. The calibration curve showed the good calibration ( P=0.125). Conclusion:The radiomics prediction model based on unenhanced CT texture analysis have application potential for the evaluation of WHO/ISUP grading of ccRCC.

OBJECTIVETo investigate the effects of nano-lead exposure on learning and memory and iron homeostasis in the brain of the offspring rats on postnatal day 21 (PND21) and postnatal day 42 (PND42).

METHODSTwenty adult pregnant female Sprague-Dawley rats were randomly divided into control group and nano-lead group. Rats in the nano-lead group were orally administrated 10 mg/kg nano-lead, while rats in the control group were administrated an equal volume of normal saline until PND21. On PND21, the offspring rats were weaned and given the same treatment as the pregnant rats until 42 days after birth. The learning and memory ability of offspring rats on PND21 and PND42 was evaluated by Morris water maze test. The hippocampus and cortex s amples of offspring rats on PND21 and PND42 were collected to determine iron and lead levels in the hippocampus and cortex by inductively coupled plasma-mass spectrometry. The distributions of iron in the hippocampus and cortex were observed by Perl's iron staining. The expression levels of ferritin, ferroportin 1 (FPN1), hephaestin (HP), and ceruloplasmin (CP) were measured by enzyme-linked immunosorbent assay.

RESULTSAfter nano-lead exposure, the iron content in the cortex of offspring rats on PND21 and PND42 in the nano-lead group was significantly higher than those in the control group (32.63 ± 6.03 µg/g vs 27.04 ± 5.82 µg/g, P<0.05; 46.20 ±10.60 µg/g vs 36.61 ± 10.2µg/g, P<0.05). The iron content in the hippocampus of offspring rats on PND42 in the nano-lead group was significantly higher than that in the control group (56.9 ± 4.37µg/g vs 37.71 ± 6.92µg/g, P<0.05). The Perl's staining showed massive iron deposition in the cortex and hippocampus in the nano-lead group. FPNl level in the cotfex of offspring rats on PND21 in the nano-lead group was significantly lower than that in the control group (3.64 ± 0.23 ng/g vs 4.99 ± 0.95 ng/g, P<0.05). FPN1 level in the hippocampus of offspring rats on PND42 in the nano-lead group was significantly lower than that in the control group (2.28 ± 0.51 ng/g vs 3.69 ± 0.69 ng/g, P<0.05). The escape latencies of offspring rats on PND21 and PND42 in the nano-lead group were longer than those in the control group (15.54 ± 2.89 s vs 9.01 ± 4.66 s; 6.16 ± 1.42 s vs 4.26 ± 1.51 s). The numbers of platform crossings of offspring rats on PND21 and PND42 in the nano- lead group were significantly lower than those in the control group (7.77 ± 2.16 times vs 11.2 ± 1.61 times, P<0.05; 8.12 ± 1.51 times vs 13.0 ± 2.21 times, P<0.05).

ONCLUSIONn Nano-lead exposure can result in iron homeostasis disorders in the hippocampus and cortex of offspring rats and affect their learning and memory ability.

Animals ; Cerebral Cortex ; drug effects ; metabolism ; Female ; Hippocampus ; drug effects ; metabolism ; Homeostasis ; Iron ; metabolism ; Lead ; toxicity ; Learning ; drug effects ; Maternal Exposure ; adverse effects ; Memory ; drug effects ; Pregnancy ; Rats ; Rats, Sprague-Dawley

Objective To establish cell strain expressing the genes of HIV vpr and mutant HIV vpr-FS, and to explore cell apoptosis ability by HIV Vpr and Vpr-FS. Methods The recombinant plasmids were constructed by cloning HIV vpr and HIV vpr-FS genes into the eukaryotic expression vector pcDNA3.1respectively. To determine the primary structures of HIV vpr and HIV vpr-FS, plasmids were cleaved by restriction enzymes. After the plasmids were transfected into HeLa cells by liposome, the HeLa cells were selected with G418 selective medium, mRNA expression of HIV vpr or HIV vpr-FS of transfected cells was detected by RT-PCR, and Vpr and Vpr-FS protein expression were detected by Western blot assay respectively. The DNA content and the percentage of apoptosis in HeLa HIV vpr cell, HeLa HIV vpr-FS cell and HeLa pcDNA3.1 cell were monitored by flow cytometry and the DNA fragmentation was analyzed by agarose gel electrophoresis. Results BamH Ⅰ and Hind Ⅲ cleavaged products of pcDNA3.1-vpr and pcDNA3.1-vpr-Fincluded 342 bp length fragments suggesting that the length of DNA sequence containing HIV vpr (HIV vpr-FS) within pcDNA3.1 was the same as theoretical length. The HeLa cells transfected by pcDNA3.1-vpr or pcDNA3, l-vpr-FS and selected with G418 could express HIV vpr or HIV vpr-FS by RT-PCR, and express HIV Vpr or HIV Vpr-FS protein by Western blot. The results of flow cytometry and DNA fragmentation showed that there was significant different in the number of apoptotic cells between HeLa HIV vpr cell and HeLa HIV vpr-FS cell, but the difference between HeLa HIV vpr-FS cell and control group was not obvious. Conclusion Recombinant plasmids pcDNA3.1-vpr and pcDNA3. 1-vpr-FS were constructed successfully, and the cell strain expressing HIV Vpr and HIV Vpr-FS proteins was established. The HIV Vpr could induce host cell apoptosis, while the mutant of Vpr did not or weakened this ability. This study provides foundation for further study on HIV vpr gene.

This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.

This article was aimed to study immunomodulatory effect of chemical split fractions ofMori Cortex, in order to initially explain effective parts that played a role in immunomodulatory effect ofMori Cortex. The carbon clearance test, serum hemolysin test, E-rosette test, and lymphocyte transformation test were carried out to explore influence of these chemical split fractions ofMori Cortex on immune organs, nonspecific immunity, humoral immunity and cellular immunity. The results showed that in the carbon clearance test, 50% ethanol fraction markedly reduced the thymus index (P<0.01) and the correction indexα (P<0.05). In hemolysin test, the half value hemolysis (HC50) was improved by 30% ethanol fraction and fatty oil fraction (P<0.05). Besides, in the E-rosette test, the E-rosette ration was increased in the 30% ethanol fraction group (P<0.05). In the lymphocyte transformation test, the 30% ethanol fraction can promote the thymus and spleen lymphocytes proliferation (P<0.05 orP<0.01), while the 50% ethanol fraction inhibited the proliferation (P<0.05 orP<0.01). It was concluded that the 30% ethanol fraction can boost both the humoral immunity and cellular immunity; the 50% ethanol fraction can induce the growth of thymus with a suppressive effect on nonspecific immunity and cellular immunity; the fatty oil fraction can improve humoral immunity.

This study was aimed to investigate the antitussive, expectorant and antiasthmatic effects of aqueous extracts and the chemical split fractions ofMori Cortex. Cough mice models induced by ammonia water were used to observe the effect on arresting cough. The phenol red expectoration method in mice was used to observe the effect on expelling phlegm. Histamine and acetylcholine mixture induced asthma model was used to observe the effect on relieving asthma. Isolated trachea model was used to observe the effect on relieving spasm. Compared with the control group, the low, medium and high doses of aqueous extracts ofM. Cortex can obviously decrease the frequency of cough, increase phenol red output of trachea in mice, prolong the latent period of asthma in guinea pigs, and increase the antispasmodic rate. The medium dose had the best effect. The comparison between different chemical split fractions ofM. Cortex and the control group showed that the 30% and 50% ethanol fraction ofM. Cortex can obviously decrease the frequency of cough and prolong the latent period of cough induced by ammonia water in mice, increase phenol red output of trachea in mice (P<0.05 orP<0.01); and 30% ethanol fraction and fatty oil fraction can prolong the latent period of asthma in guinea pigs (P< 0.05 orP<0.01). In addition, 30% ethanol fraction can obviously reduce the degree of tracheal contraction. It had certain effect of relieving spasm. It was concluded that aqueous extracts ofM. Cortex had better effects on relieving cough, expectorant and antiasthma. The effective part was the 30% ethanol fraction, which was the dose equivalent of 1/3 of the medium dose. It had significant effect on relieving cough, expectorant and antiasthma. The effect was equivalent to the medium dose of aqueous extracts of M. Cortex.