Objective To separate the side population cells(SP) from breast cancer MCF-7 cell line,and observe its biological characteristics.Methods Flow cytometry and Hcechst 33342 dye efflux assay were used to isolate SP cells and non-SP cells from the MCF-7 cell line of human breast cancer.Tumorigenicity of the two subpopulations was observed by a soft agar cloning method.Results The results of FACS analysis indicated that (6.5 ± 0.4 ) ％of the MCF-7 cells were SP cells;The vitro colony formation rate of SP cells was(38.5 ±9.4)％,and higher than that of non-SP cells ( 8.4 ± 2.6 ) ％ ( t =5.34,P ＜ 0,05 ).Concluslon The SP cells sorted from MCF-7 cell line enriched tunor stem cells,which exhibited high tumorigenicity.It indicated that SP cells should play a principal role in breast cancer.

Adolescent ; Biopsy ; Female ; Hepatocytes ; diagnostic imaging ; pathology ; Humans ; Jaundice, Chronic Idiopathic ; pathology ; Sibling Relations ; Ultrasonography

This study is to investigate the influence and mechanism of action of asymmetrical dimethylarginine (ADMA) and the induced oxidative stress level on Alzheimer's disease (AD) incidence. ADMA concentration, nitric oxide, Abeta(40)/Abeta(42) ratio, inducible NO synthase (iNOS) activity and the concentrations of the induced free radicals including malondialdehyde (MDA), 3-nitrotyrosine (3-NT) and peroxynitrite (ONOO-) in the cerebrospinal fluid (CSF) from 34 neurologically normal controls and 37 AD patients were quantitatively determined and statistically compared. The results showed that the ADMA concentration significantly decreased in AD patients, and it showed negative correlation with the NO, iNOS activity, and showed positive correlation with MMSE score. ADMA concentration was negatively correlated with Abeta(40)/Abeta(42) ratio (P<0.01) with the observation that Abeta(40)/Abeta(42) ratio increased while ADMA level decreased in CSF in AD patients. The concentration levels of MDA, 3-NT and ROS significantly increased compared with the control with all the P values less than 0.05. These findings suggested that the ADMA disorder and the oxidative damage effect of the induced free radicals in CSF of AD patients are an important mechanism of AD incidence, and their joint regulation may provide new idea for the prevention and clinical treatment of AD.

The secondary metabolites of plants are important sources of natural drugs. Betula plants have abundant pharmacological value, complex mechanism and wide applications, which are closely related to the triterpenoids of theirs. Triterpenoids in Betula species are mainly divided into dammarane-type, ocotillol-type, oleanane-type, lupane-type and cycloaltunane-type. The extracts of Betula species have varieties of activities such as anti-tumor, anti-inflammatory, anti-oxidant, anti-bacterial, etc. And the biosynthetic pathways of triterpenoids after 2,3-oxidosqualene are split into four branches of dammarenediol-II, lupeol, cycloartenol and amyrin according to the different oxidosqualene cyclases. This review summarizes the chemical constituents, pharmacological activities and biosynthetic pathways of triterpenoids in Betula plants. It provides a reference for the research and development of new drugs and the production of these triterpenoids in microbial cell factories by synthetic biology methods.

Natural products are important sources of drug discovery. However, the traditional methods of extraction and isolation, as well as chemical synthesis for obtaining natural products are associated with issues such as operational complexity, high costs, low efficiency, and environmental pollution. Constructing microbial cell factories through synthetic biology methods to produce medicinal natural products has the advantages of high efficiency, low cost, and environmental protection. Nevertheless, the scope and yield improvement of the products are limited by the limitations of enzymes in microbial cell factories. Protein engineering is considered one of the most effective approaches to overcome these limitations. This article introduces commonly used methods of protein engineering technology and summarizes its specific applications in improving enzyme performance, modifying the enzymatic environment, and promoting the development of synthetic biology tools in the field of pharmaceutical natural product synthesis. Furthermore, it analyzes the current bottlenecks and challenges in protein engineering and looks forward to its future application prospects, offering insights for the development and practical use of protein engineering technology.

Objective To discuss the combined value of gray-scale ultrasound ( GSUS) and contrast-enhanced ultrasound (CEUS) in the diagnosis of benign and malignant thyroid lesions and to explore the optimal diagnostic point of scoring method .Methods Ultrasound images of 178 thyroid lesions confirmed by pathology were synthetically reviewed by scoring 5 GSUS indicators including shape , orientation, interior echogenicity, halo sign, microcalcification and 6 CEUS indicators including relative arrival time of microbubbles in the periphery and interior, peak periphery and interior echogenicity, peripheral ring-enhancement, and homogeneity of enhancement .One positive indicator scored one point .The optimal diagnostic points and their clinical value were explored according to ROC curves .Results Scores of GSUS, CEUS and the combination of GSUS and CEUS were significantly different (Z =10.188,9.843,10.705,all P <0.001). Areas under ROC curves of GSUS, CEUS and the combination of GSUS and CEUS were 0.936, 0.919 and 0.964, respectively.Three or more positive GSUS indicators of five in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 79.6% and the specificity of 91.2%.Two or more positive CEUS indicators of six in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 91.8% and the specificity of 81.2%.Five or more positive GSUS and CEUS indicators of eleven in a thyroid lesion predicted that the thyroid lesion was malignant , with the sensitivity of 93.6%and the specificity of 92.3%.The areas under the ROC curve of GSUS and CEUS were 0.936 and 0.919.The area under the ROC curve of the combination of GSUS and CEUS was 0.964, larger than the areas under the GSUS ROC curve and the CEUS ROC curve.Conclusion Ultrasound is valuable in the differential diagnosis of benign and malignant thyroid lesions , and the combination of GSUS and CEUS is the most valuable with 5 points as the optimal diagnostic scoring method .

OBJECTIVE To observe the clinical effects of the combination of Duhuo Bugu decoction and zoledronic acid in the treatment of malignant bone metastasis.METHODS Fifty patients with bone metastases of malignant tumor were selected from the Department of oncology,Chongqing Hospital of traditional Chinese medicine from September 2014 to February 2016, and were randomly divided into the observation group and the control group.Two groups were both treated by zoledronic acid, and the observation group was also treated by adding Duhuo Bugu Decoction.Changes of the analgesic effects,analgesic dosa-ges and life quality scores(KPS)of two groups were observed before and after treatment.Urinary type I collagen amino termi-nal peptide(uNTX)and bone alkaline phosphatase(BALP)were measured via ELISA method before and after treatment.RE-SULTS Compared with the control group,the analgesic medicinal amount of the observation group reduced significantly(P<0.05),and KPS improved significantly(P<0.05).After the treatment,numerical pain scores(NRS) of two groups had no significant changes(P>0.05).After the treatment,uNTX and sBALP level of the treatment group decreased compared with that before treatment(P<0.01).While compared with the control group,the difference was not significant(P>0.05).CON-CLUSION The combination of Chinese herbs and zoledronic acid is an effective therapy for bone metastasis malignant tumors. Chinese herbs can effectively improve the analgesic effect,reduce the analgesic medicinal amount,improve the patients'life quality,and have a tendency of synergy in improving the bone destruction caused by tumor.

Aim To evaluate the vasorelaxant effect of two new chemical entities, J35242 and J35243, on iso-lated rat thoracic aorta rings as Rho-kinase inhibitors, and further to explore the underlying mechanisms of these two compounds. Methods Isolated rat thoracic aorta rings pre-contracted by KCl or norepinephrine ( NE) were used to evaluate the vasodilatory effect of J35242 and J35243 . Through the interventions of sev-eral tool drugs, the mechanisms of compounds concern-ing endothelium, K+ channels and Ca2+ were studied. Results J35242 and J35243 showed potent relaxant effect on both KCl and NE pre-contracted vessels, and exhibited partial endothelium dependency. L-NAME and Methylene Blue( MB) could influence the relaxant effect of these compounds. Meanwhile, the compounds could inhibit intracellular Ca2+ release and extracellu-lar Ca2+ influx, which indicated that the compounds might block the calcium channels to relax the vessels. In addition, the two compounds probably did not dilate the aorta rings through opening potassium channels. Conclusions J35242 and J35243 have vasorelaxant effects on vessels in vitro and the potency of J35242 is stronger than that of J35243 . The underlying mecha-nisms might be endothelium-dependent. Also the com-pounds might block Ca2+ channels, lowering intracel-lular Ca2+ concentration to relax the vessels.

Objective To compare the effect on intraoperative cerebral metabolism between the propofol combined with remifentanil infusion in total intravenous anesthesia and the desflurane combined with remifentanil intravenous inhalation anesthesia in patients in neurosurgery.Methods Thirty-four patients were randomly divided into the propofol combined with remifentanil group (group A,n=16) and the desflurane combined with remifentanil group (group B,n =18).The B-ultrasound under the guide of retrograde catheterization through right internal jugular vein and artery was prepared after the patients entered the operation room.Atropine,propofol,fentanyl,rocuronium were used in the induction of anesthesia.The mechanical ventilation was applied after conventional trachea cannula.Once patients were anesthetized steadily,jugular bulb venous oxygen saturation (SjvO2),jugular bulb venous oxygen partial pressure (PjvO2),Jugular Bulb venous hemoglogin (Hbv),jugular bulb venous lactate (Ljv),jugular bulb venous gluxose (Gv),arterial oxygen saturation (SaO2),arterial oxygen pressure (PaO2),arterin (Hba),arterial blood lactate (La),arterial blood gluxose (Ga),arterial blood oxygen content (CaO2),jugular bulb venous oxygen content (CjvO2),arteriovenous oxygen difference (AVDO2),cerebral extraction of oxygen (CEO2),cerebral lactate production rate (CLP) and cerebral glucose uptake rate (CGU) at different time [before anesthesia induction (T1),1 hours after the start of the operation (T2),2 hours after the start of the operation (T3),half an hour after the operation]were collected.Results (1) The value of SjvO2,PjvO2,CjvO2 and CaO2 in group B was significantly higher than that in group A (P＜0.001);The value of AVDO2 and CEO2 in group B was lower than that in group A (P＜0.05);(2) The value of Gv and Ga in group B was higher than that in group A (P＜0.05);There were no significant differences about CGU in two groups (P＞0.05);(3) The value of CLP in group B was lower than that in group A (P＜0.05);there were no significant differences about Ljv and La in two groups (P＞0.05);(4) Compared with the value at moment of T1 between group A and group B,the value ofPaO2,SaO2,PjvO2 and SjvO2 were increased with time (P＜0.05),the value of CaO2,CjvO2,AVDO2 and CEO2 showed a downward trend (P ＜0.05).Conclusion (1) Both total intravenous anesthesia and intravenous inhalation anesthesia can reduce the cerebral oxygen metabolism;(2) For the cerebral protection of neurosurgery operation,it seems that the effect of intravenous inhalation anesthesia is more stronger than total intravenous anesthesia.

OBJECTIVETo investigate whether programmed death ligand 1 (PD-L1) expressed in the periodontal tissue of chronic periodontitis and the correlativity of PD-L1 and different degrees of chronic periodontitis, provide experience for immunoregulation mechanism, clinical treatment and prognosis of chronic periodontitis.

METHODSGingiva and periodontal tissue of healthy people and chronic periodontitis patients were collected. Based on clinical probing, periodontal tissue were classified into three groups: periodontal tissues of healthy people, periodontal tissue of mild chronic periodontitis, periodontal tissue of severe chronic periodontitis. Fluorescent quantitation polymerase chain reaction was applied to explore the expression of PD-L1 mRNA in the periodontal tissue of the different groups. Western blot and immunohistochemistry method were utilized to test the expression of PD-L1 protein in the periodontal tissue of the different groups. Combining with clinical image data, the relationship between differentially expressions of PD-L1 and different degrees of chronic periodontitis was analyzed.

RESULTSThe relative expression quantity of PD-L1 in the periodontal tissue of the mild chronic periodontitis was significantly higher that of the severe chronic periodontitis (P<0.01). The relative expression quantity of PD-L1 in the periodontal tissue of healthy subjects and severe chronic periodontitis had no statistical significance (P>0.05).

CONCLUSIONThe expression of PD-L1 in the periodontal tissue negativelv regulates inflammatory periodontal tissue damage.