Body Fluids ; cytology ; Cytodiagnosis ; methods ; Humans ; Immunohistochemistry ; methods ; Lung Neoplasms ; pathology ; Pleural Effusion, Malignant ; pathology ; Quality Control ; Specimen Handling ; standards

As an important gene regulatory molecule, microRNA is closely related with various human diseases. Numerous studies have confirmed that microRNAs function as tumor suppressors or oncogenes in various tumor types. Therefore, microRNA investiga-tion has become a new direction in oncology research. As a member of the miR-17 to-92 cluster, miR-17-5p has been the focus of re-search recently. MicroRNA is involved in many aspects of diseases, such as diabetes mellitus, endometriosis, and a variety of tumors. In this review, the basic characteristics and roles of miR-17-5p in tumors are elaborated.

Objective To obtain highly expressing cell lines by inserting the glutamine synthetase (GS) screening system and replacing the promoter of the vector.Methods The mutation of the point BamHⅠwas induced to build a new vector pIRES2-EGFP.The marker gene GS was inserted by AseⅠ and NheⅠ, and the promoter hCMV was replaced by PacⅠand NheⅠ.The new vector pHGS1.0 and the vector pIRES2-enhanced screen fluorescein protein( EGFP)-B were inserted by the recombinant protein TEM8 ( 1-227 )-VEGFR1 domain2-IgG2 ( TV-IgG2 ) gene to analyze the advantages of the expression.Results The glutamine synsthetase is successfully inserted, the human cytomegalovirus replaced, and recombinant protein is increased 5-fold by human immunoglobulin quantification kit.Conclusion The GS system is a highly protein expressing system.

Objective To understand the status quo of the illness situation and the implementation of prevention and control measures on Kashin-Beck disease( KBD ) in Shandong Province , and provide a scientific basis for assessment and acceptance of the work on eliminating KBD. Methods In accordance with the requirements of the national “Implementation Scheme for Endemic Disease Control in 2012” and the “Shandong Provincial Technical Scheme for KBD Control in 2012”. In four towns of Qingzhou, three epidemic villages were selected as the surveying villages in each town in order to understand the implementation of measures for prevention and control of KBD; information such as population of investigating villages, per capita income, farming, food and the implementation of control measures,etc were included. In each investigating village, 7 - 12 years old children were selected to receive clinical and X-ray examination. When fewer than 50 people in a investigating village , one additional endemic village was included near the investigating village , and all children aged 7 to 12 of the village were checked. The number of cases of clinical degree Ⅰ, Ⅱ, Ⅲand the detection rates, X-ray positive detection rate, the number of cases of metaphysis, epiphysis, distal end of phalanx, carpal bone, triad and the detection rates were calculated, respectively. Clinical and X-ray diagnosis were based on “The Diagnostic Criteria of Kaschin-Beck disease”(WS/T 207-2010). Results A total of 14 epidemic villages were selected to survey in four towns of Qingzhou. In the recent three years in the epidemic areas, measures of supplying Se had not been implemented;measures of resettlement and off-site education had not been implemented either. In the epidemic areas , the main crops were wheat and corn; the staple food was flour which accounted for more than 80%, and the foreign food bought accounted for only 11%. The areas of returning farmland to forest (grass) accounted for 0.32%(36/11 151) of the original arable land, and the areas of replanting economic crops accounted for 1.17%(131/11 151) of the original farmland. A total of 536 children aged 7 to 12 were examined by clinical and X-rays. Patient of KBD was not detected clinically. Six positive patients with metaphyseal changes of KBD were detected by X-rays, and the average X-ray detection rate was 1.12%(6/536). Patients with abnormal epiphysis, distal end of phalanx and carpal bone were not detected by X-rays. Conclusions KBD in Shandong Province has been effectively controlled. Since the cause of KBD is still unknown, it is necessary to build a sustainable long-term control mechanism of KBD;further improve the monitoring system and dynamic monitoring work still need to be strengthened.

Objective To observe the effect of APA-BCC(alginate-polylysine-alginate microencapsulated bovine adrenal medullary chromaffin cells)microcapsules transplantation into the subarachnoid space of cancer pain patients on endogenous opioid peptides and catecholamine concentration in cerebrospinal fluid(CSF).Methods The different doses of APA-BCC microcapsules were transplanted into the spinal subarachnoid space of cancer patients with moderate or serious pain.The concentrations of Leu-enkephalin(L-EK),β-endorphin(β-EP),dynorphin A(DynA),noradrenaline(NA)and adrenaline(AD)in CSF were tested.Results L-EK concentration in CSF increased remarkably after transplantation,and the increase was most remarkable when transplantation doses were 1.0×107and 1.25×107;there were no remarkable changes of β-EP,DynA,NA and AD after transplantation.Conclusion The analgetic effects of APA-BCC microcapsules tranplantation may correlate with the increase of L-EK in CSF of cancer pain patients;the dose of 1.0×107 and 1.25×107cells may be the most effective dose.

Intraperitoneal administration of timosaponin A-III (TA-III) has therapeutic effects on high-fat diet-induced metabolic dysfunction-associated steatotic liver disease (MASLD), but oral administration has no effect. This suggests that gut microbiota may affect the oral bioavailability of TA-III. Metabolic dysfunction-associated steatohepatitis (MASH) is an inflammatory subtype of MASLD. To investigate the therapeutic effect of different administration modes of TA-III on MASH and its relationship with gut microbiota metabolism. In this study, a MASH mouse model was induced by choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD). Comparing the therapeutic effect of intraperitoneal injection (10 mg·kg^-1, ip) and intragastric administration (100 mg·kg^-1, ig) of TA-III. The concentration of TA-III in serum of rats under the two administration modes was analyzed. On this basis, the metabolic effect of gut microbiota on TA-III in mice was verified by the experiment of metabolism of gut microbiota in vitro. The pharmacokinetic experiment of combined antibiotic intervention in mice further verified the metabolism of TA-III by gut microbiota in mice. Finally, the concentration of TA-III in serum of mice after the administration of TA-III by intragastric administration under different antibiotic intervention conditions was compared, and 16S rRNA sequencing analysis was combined to find the key bacteria that may participate in the metabolism of TA-III. The animal welfare and experimental procedures in this paper were in accordance with the provisions of the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine. The ethics approval number is PZSHUTCM2307030004 and PZSHUTCM2310200003. The results showed that TA-III (10 mg·kg^-1, ip) had definite therapeutic effect on MASH mice, but TA-III (100 mg·kg^-1, ig) was ineffective. The analysis showed that the prototype concentration of TA-III in serum and liver of mice in TA-III (100 mg·kg^-1, ig) was significantly lower than TA-III (10 mg·kg^-1, ip), suggesting that the oral administration of TA-III may be metabolized by gut microbiota. The concentration of TA-III in serum of streptomycin (Str) treated mice was higher than normal mice. Combined with 16S rRNA gene sequencing analysis, it was found that the abundance of Akkermansia_muciniphila (A. muciniphila) was significantly reduced in the Str group. In vitro experiments showed that A. muciniphila could metabolize TA-III. In conclusion, gut microbiota is an important factor affecting the efficacy of TA-III administration through the gastrointestinal tract, in which A. muciniphila may play an important role.

OBJECTIVE To investigate the chemical constituents of the roots and rhizomes of Ligularia macrophylla.METHODS The compounds were isolated through repeated column chromatography on silica gel, polyamide, RP-18 and Sephadex LH-20. And the structures were elucidated on the basis of spectroscopic data (UV, IR, MS, 1H NMR, 13C NMR, DEPT and 2D NMR). RESULTS Four flavonol glycosides and two monoterpene glycosides were obtained and determined as rhamnazin-3-O-β-D-rutinoside ( 1 ), rhamnetin-3-O-β-D-rutinoside ( 2), rutin ( 3), afzelin ( 4), betulalbuside A (5) and 3,7-dimethyloct-1 -en-3,8-diol8-O-β-D- glucopyranoside (6). CONCLUSION All the compounds were isolated from this plant for the first time.

Cell Culture Techniques ; methods ; Fixatives ; Humans ; Immunohistochemistry ; methods ; Tissue Fixation ; methods

OBJECTIVETo develop a novel method for preparing decellularized liver biological scaffold (DLBS) for liver tissue engineering.

METHODSDLBS was prepared by treatment of rat livers with detergent and enzymatic cell extraction and observed under optical and scanning electron microscopes. To assess the biocompatibility of the product, C3A cells and bone marrow-derived mesenchymal cells (BM-MSCs) were cocultured with DLBS as the scaffold, and the effect of DLBS on the proliferation of C3A cells was evaluated by MTT assay. DLBS was also implanted under the dorsal skin of SD rats to evaluate the tissue biocompatibility of this material.

RESULTSApplication of the detergent and enzymatic extraction allowed full extraction of the cells in the liver, leaving an extracellular matrix scaffold composed mainly of collagen and elastic fibrin. The coculture experiment showed that C3A cells and BM-MSCs could grow on and adhere to DLBS. The result of MTT assay showed that DLBS could promote the proliferation of C3A cells.

CONCLUSIONThis cell-free DLBS, which retains intact extracellular matrix and promotes cell attachment, proliferation, growth and differentiation, can be an ideal biological matrix scaffold material.

Animals ; Biocompatible Materials ; Cells, Cultured ; Coculture Techniques ; Extracellular Matrix ; chemistry ; Female ; Liver ; cytology ; Male ; Mesenchymal Stromal Cells ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

ObjectiveTo investigate the current status of Kaschin-Beck disease in Shandong province, and to provide a scientific basis for decision-making in controlling the disease. Methods According to the National Monitoring Program of Kaschin-Beck disease requirements, historical serious villages of Kaschin-Beck disease in Qingzhou of Shandong province were selected annually; children aged 7 to 16 were chosen to receive clinical examination and children aged 7 to 12 were taken X-ray examination. Clinical and X-ray diagnosis was carried out according to the Diagnostic Criteria of Kashin Beck Disease(GB 16003-1995). Results From 1996 to 2010, in 53 diseased villages, three thousand three hundred and eighteen school children aged 7 to 16 were clinically diagnosed, and child Kaschin-Beck disease of degree Ⅰ and above were not detected; three thousand and ninety-one school children aged 7 to 12 were examined by X-ray, forty cases were found positive, and the total positive rate was 1.29％(40/3091 ). The year with the highest positive rate was 2002, and the rate was 3.49％(13/372) ; the positive rate was 0 in 1996 and 2008. The difference of the X-ray positive rate between each year was statistically significant(x2 =31.54, P ＜ 0.01 ). ConclusionsChild Kashin-Beck disease in Qingzhou is basically under control.Since etiology of Kashin-Beck disease is still unclear, surveillance of the disease still needs to be strengthened.