Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant metabolites renowned for their pharmacological properties. However, sustainable sources for these compounds remain limited. Consequently, researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches. CYP80B, a family of cytochrome P450 (CYP450) enzymes, plays a crucial role in BIA biosynthesis. Previously reported CYP80Bs are known to catalyze the 3'-hydroxylation of (S)-N-methylcoclaurine, with the N-methyl group essential for catalytic activity. In this study, we successfully cloned a full-length CYP80B gene (StCYP80B) from Stephania tetrandra (S. tetrandra) and identified its function using a yeast heterologous expression system. Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that StCYP80B could catalyze N-methylcoclaurine and coclaurine into their respective 3'-hydroxylated products. Notably, StCYP80B exhibited an expanded substrate selectivity compared to previously reported wild-type CYP80Bs, as it did not require an N-methyl group for hydroxylase activity. Furthermore, StCYP80B displayed a clear preference for the (S)-configuration. Co-expression of StCYP80B with the CYP450 reductases (CPRs, StCPR1, and StCPR2), also cloned from S. tetrandra, significantly enhanced the catalytic activity towards (S)-coclaurine. Site-directed mutagenesis of StCYP80B revealed that the residue H205 is crucial for coclaurine catalysis. Additionally, StCYP80B exhibited tissue-specific expression in plants. This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
Cytochrome P-450 Enzyme System/chemistry* ; Benzylisoquinolines/chemistry* ; Hydroxylation ; Plant Proteins/chemistry* ; Alkaloids/metabolism* ; Stephania tetrandra/genetics*

Objective To investigate the role of knockdown of peroxiredoxin-6(PRDX6)in injury and adaptive expression of bile acid transporter in human hepatoellular carcinomas(HepG2)cells induced by rifampicin(RFP).Methods Cells in logarithmic growth phase were uniformly inoculated in six-well plates,and HepG2 cells were transiently transfected with specific PRDX6-siRNA and control-siRNA to construct the knockdown group and control group.After 24 h of induction with 100 μmol/L RFP,Western blot and qRT-PCR were performed to detect the protein and gene expression levels of PRDX6,multidrug resistance protein 1(MDR1),multidrug resist-ance-associated proteins 2,3 and 4(MRP2,MRP3 and MRP4),and Na+/taurine taurocholate cotransporter pro-tein(NTCP).Annexin V-FITC/PI double staining assay was used to detect the apoptosis rate of cells in each group;CCK-8 assay was used to detect the changes of cell proliferation in each group;The relative contents of ala-nine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),indirect bilirubin(IBIL)and total bile acid(TBA)in the supernatant of cell culture medium of each group were detected by kits.Results RFP increased the protein and gene expression levels of MRP2,MRP3,MRP4,MDR1,NTCP and PRDX6 in HepG2 cells(P<0.05),while the protein and gene expression levels of MRP2,MRP3,MRP4,MDR1 and NTCP decreased to different degrees after PRDX6 knockdown(P<0.05).In addition,PRDX6 knockdown re-sulted in increased apoptosis rate of HepG2 cells(P<0.05),decreased cell proliferation ability(P<0.05),and increased levels of cell injury markers(ALT,AST,TBIL,DBIL,TBA)in cell culture supernatants(P<0.05).Conclusion RFP increased the protein and gene expression of bile acid transporter and PRDX6 to increase in HepG2 cells.However,following knockdown of PRDX6 and treatment with RFP,the protein and gene expression levels of the bile acid transporter decreased and cell injury was aggravated,suggesting that PRDX6 played a protec-tive role in RFP-induced adaptive response in HepG2 cells.

Objective : To investigate the effect of mesencephalic astrocyte-derived neurotrophic factor (MANF) on the adaptive expression of bile acid transporter in human hepatoellular carcinomas (HepG2) induced by rifampicin (RFP) ． Methods: The control group cell line (Y07) and the knockdown group cell line (Y25) were constructed by lentiviral stable transfection technology．The Y07 and Y25 cells were treated with RFP of 200 μmol / L for 48 h， and qRT-PCR and Western blot were used to detect the protein and gene expression levels of MANF，bile salt export pump ( BSEP) ，multidrug resistance-related proteins 2 /3 /4 ( MRP2 ，MRP3 ，MRP4) ，multidrug resistance protein 1 (MDR1) ，organic solute transporter a / β ( OSTα/ β) ，organic anion transporter ( OATP2B1) ．The protein and gene expression levels of proliferating cell nuclear antigen ( PCNA) ，proliferating cell marker Ki67 were used to evaluate the proliferation of cells in each group changes in levels．Changes in the protein and gene expression levels of C / EBP homologous protein( CHOP) and cysteinyl aspartate specific proteinase-3 ( Caspase-3) were used to evaluate the apoptosis of cells in each group．The relative contents of alanine aminotransferase(ALT) ，aspartate aminotransferase(AST) ，alkaline phosphatase ( ALP) ，total bilirubin ( TBIL) ，indirect bilirubin ( IBIL) and total bile acid(TBA) in the supernatant of cell culture medium of each group were detected by kits. Results: RFP could induce the protein and gene expression of MANF，BSEP ，MRP2 ，MRP3 ，MRP4 ，MDR1 ，OSTα , OSTβ , OATP2B1 in HepG2 cells (P ＜0. 05 ) ，while the protein and gene expression levels of BSEP ，MRP2 ，MRP3， MRP4，MDR1，OSTα、OSTβ、OATP2B1 decreased after MANF knockdown(P＜0. 05) ．Moreover，under the action of RFP，the protein expression of PCNA and Ki67 in the knockdown group was still higher．The protein and gene levels of CHOP and Caspase-3 significantly increased after MANF knockdown(P＜0. 05) ．The levels of the hepatic cell injury markers in the cell supernatant increased significantly(P＜0. 05) ． Conclusion RFP can induce the expression of bile acid transporter such as BSEP，MRP2，MRP3，MRP4，MDR1，OSTα , OSTβ and OATP2B1 to increase in HepG2 cells(P＜0. 05) ，but the expression of bile acid transporter of HepG2 after MANF knockdown will significantly decrease under the induction of rifampicin(P＜0. 05) ，and cell indury is aggravated，indicating that MANF plays a protective role in RFP-induced adaptive responses by regulating the bile acid transporter.

ABSTRACT: Meningiomas are the most common primary intracranial neoplasm with diverse pathological types and complicated clinical manifestations. The fifth edition of the WHO Classification of Tumors of the Central Nervous System (WHO CNS5), published in 2021, introduces major changes that advance the role of molecular diagnostics in meningiomas. To follow the revision of WHO CNS5, this expert consensus statement was formed jointly by the Group of Neuro-Oncology, Society of Neurosurgery, Chinese Medical Association together with neuropathologists and evidence-based experts. The consensus provides reference points to integrate key biomarkers into stratification and clinical decision making for meningioma patients. REGISTRATION Practice guideline REgistration for transPAREncy (PREPARE), IPGRP-2022CN234.
Humans ; Meningioma/pathology* ; Consensus ; Neurosurgical Procedures ; Meningeal Neoplasms/pathology*

Objective To obtain the information on the frequency of diagnostic X-ray examinations in Guangdong province in order to provide reference data for the rational application of diagnostic radiology.Methods A typical stratified sampling method was used to select 56 medical institutions in 7 districts in Guangdong province for the survey of the frequencies of diagnostic X-ray examinations.Multiple linear regression analysis was performed using the relevant variables of the surveyed sample (total number of hospitals,number of outpatients,number of inpatients,number of radiation workers,and number of diagnostic equipment).The dependent independent variables were fitted to the linear regression equation to estimate the frequency of routine diagnostic X-ray examinations and diagnostic CT examinations and the total frequency of diagnostic medical X-ray examinations in Guangdong province in 2016.Results By the largest percentage contribution,chest radiography accounted for 60.84％ of the routine diagnostic X-ray examinations,brain scans 28.03％ of diagnostic CT examinations,and cardiovascular interventions 48.83％ of interventional radiology.For the group of above 40 years old,the largest percentage contributions were,respectively,routine diagnostic X-ray examination 47.95％,diagnostic CT examinations 65.41％,and interventional radiology 75.64％.All types of medical X-ray examinations are concentrated in the tertiary and secondary hospitals.Based on this survey,the examination frequencies for 2016 in Guangdong were estimated to be,respectively,diagnostic X-ray examination 699 examinations per 1 000 population and CT diagnostic 231 examinations per 1 000 population,with the total of 992 examinations per 1 000 population.Conclusions This survey was completed to a limited degree and less investment.The results showed that the diagnostic radiology in Guangdong province is developing with increased frequencies,with wide varieties of X-ray examination types in the tertiary and secondary hospitals.Relevant competent defpartments should pay attention to strengthening the radiation protection of examined individuals and actively providing guidance on the correct and rational applicc ation of diagnostic radiology so as to achieve profit and avoid disadvantages.

Objective To investigate the behavioral characteristics of circadian rhythm disturbance in 3xTg-AD mice.Methods The free-running period,average activity per hour,total amount of exercise and circadian amplitude were examined with voluntary wheel-running test in 3-to 9-month-old C57BL/6 and 3xTg-AD mice under 12 h light/12 h dark cycle and constant darkness environment.Results In constant darkness environment,the free-running period in 3-month-old 3xTg-AD mice was (23.2±0.4) h,and shorter than the period((23.5±0.2) h) in C57BL/6 mice.In 6-and 9-month-old 3xTg-AD mice,activity pattern was disorganized,without clear boundary between activity and rest phase.The free-running period was unavailable.The circadian amplitude and total exercise amount were(40.6± 11.5) counts/5 min and(2.6±0.1) × 104 counts/d,(37.0±20.8) counts/5 min and(2.3±0.4) × 104 counts/d,(29.3± 11.0) counts/5 min and(1.6± 0.9) × 104 counts/d in 3-,6-and 9-month-old 3xTg-AD mice,respectively,which was significantly lower than that in age-matched C57BL/6 mice.In 12 h light/12 h dark cycle,the circadian amplitude and total exercise amount of 3-,6-and 9-month-old 3xTg-AD mice were (87.0 ± 37.8) counts/5 min and (2.2 ± 0.8) × 104 counts/d,(25.9± 6.3) counts/5 min and (1.1 ± 0.2) × 104 counts/d,(14.3 ± 5.7) counts/5 min and (0.6 ± 0.3)× 104 counts/d respectively,and with a significant decrease from the age of 6 months.Meanwhile,the locomotor activity decreased at night and increased during the day.Conclusion The endogenous circadian rhythm disturbance emerges in 3xTg-AD mice at 3-month-old,while the exogenous circadian rhythm disorder appears at 6-month-old;the degree of disorder in circadian rhythm is gradually aggravated with the increase of age in of the AD mice.

Objective To investigate the effect of split-bolus spectral computed tomography(CT) on the portal venography and radiation dose.Methods The prospective study was adopted.The clinical data of 119 patients who underwent spectral CT at China-Japan Union Hopital from September 2014 to March 2015 were collected.Patients were randomly divided into the portal venography with split-bolus spectral CT single-phase enhanced scan group and portal vein multi-phase scan group by random sequence method.In the portal venography with split-bolus spectral CT single-phase enhanced scan group,the spectral CT was used with the method of split-bolus single phase imaging,and in the portal vein multi-phase scan group,standard spiral CT was used to perform three-phase scan.Two observers evaluated CT portal venography subjectively and objectively,measured CT values,contrast to noise ratio (CNR),signal noise ratio (SNR),and calculated radiation dose.Observed indices included (1) choice of optimal monochromatic images.(2) CT values of portal veins,measurement of CNR and SNR.(3) Subjective scoring of portal venography quality.(4) Comparison of radiation dose.Measurement data with normal distribution were presented as x ± s,comparison between groups was analyzed by independent sample t test.Comparison of count data was analyzed by chi-square test.Results The 113 patients were screened for eligibility,including 59 in the portal venography with split-bolus spectral CT single-phase enhanced scan group and 54 in the portal vein multi-phase scan group.(1) Choice of optimal monochromatic images:optimal monochromatic images were abstracted at 60 keV from spectral CT portal venography.(2) CT values of portal veins and measurement of CNR and SNR:the CT values of intrahepatic portal vein,extrahepatic portal vein and branches of portal vein were (319 ± 44) HU,(328 ± 53) HU,(294 ± 45) HU in the reconstructed images at the energy level of 60 keV in the portal venography with split-bolus spectral CT single-phase enhanced scan group and (213 ±41)HU,(228 ±49)HU,(210 ±41)HU in the portal vein multi-phase scan group,with significant differences between the 2 groups(t =8.04,6.34,6.82,P ＜ 0.05).The CNR of intrahepatic portal vein,extrahepatic portal vein and branches of portal vein were 15 ± 5,24 ± 8,22 ± 7 in the portal venography with split-bolus spectral CT single-phase enhanced scan group and 13 ± 4,20 ± 6,19 ± 6 in the portal vein multi-phase scan group,respectively,with no significant difference (t =-1.13,-1.89,-1.51,P ＞ 0.05).The SNR of intrahepatic portal vein,extrahepatic portal vein and branches of portal vein were 31 ± 6,29 ± 6,27 ± 6 in the portal venography with split-bolus spectral CT single-phase enhanced scan group and 29 ± 7,28 ± 9,26 ± 6 in the portal vein multi-phase scan group,respectively,with no significant differences (t =-0.688,0.615,0.600,P ＞ 0.05).(3) Subjective scoring of portal venography quality:the subjective score of image quality of portal venography was 14.3 ± 1.0 in the portal venography with split-bolus spectral CT single-phase enhanced scan group and 12.5 ± 1.8 in the portal vein multi-phase scan group,with a significant difference (t =12.43,P ＜ 0.05).(4) Comparison of radiation dose:the radiation dose was (8.1 ± 1.1)mSv of patients in the portal venography with split-bolus spectral CT single-phase enhanced scan group and (17.4 ± 7.5) mSv in the portal vein multiphase scan group,with a significant difference (t =24.14,P ＜ 0.05).Conclusion Spectral CT portal vein imaging combined with split-bolus protocol can achieve better manifestations of portal vein and its branches,and reduce radiation dose in the scanning process.

The hospital information structure, which is made up of various medical business systems, is suffering from the problems of the "information isolated island". Medical business systems in the hospital are mutually isomerous and difficult to become a whole. How to realize the internal barrier-free interaction of the patients effective medical information in the hospital and further to complete the area sharing of patients longitudinal diagnosis and treatment information has become a question having to be solved urgently in the process of healthcare informatization. Based on the HL7 standard, this paper refers to the IHE technical framework, expounds the overall structure of the interaction in the hospital internal and area sharing of medical information with the medical information exchange platform. The paper also gives the details of the whole process of the complete display of the discrete patient health information using Portal technology, which is saved in the business systems in different hospitals. It interacts internally through the information exchange platform and at last stores the information in the regional clinical data repository (CDR).
Humans ; Information Dissemination ; methods ; Medical Informatics ; organization & administration ; Medical Record Linkage

OBJECTIVETo evaluate the structural characteristics and biocompatibility of a novel nano-Ta-Ti alloy rod for its potential application in internal fixation.

METHODSTa coating of a Ti alloy rod with nano-Ta (tantalum) powder was performed using laser melting with symmetrical grooves repleted with nano-Ta powder along the whole length. The microstructure of the cross section of Ta-Ti alloy rod, pore diameter and components of the coating were analyzed using scanning electron microscopy. The influence of this nano-Ta-Ti alloy on proliferation and differentiation of MC3T3-E1 cells was evaluated by MTT cytotoxicity test and ALP activity test.

RESULTSUnder scanning electron microscope, the Ta-coating surface presented with a gross porous (200-300 µm) structure with dense fusion between Ta particles, and no new element was produced after laser melting. Biocompatibility evaluation showed that Ti alloys with and without Ta coating both promoted the proliferation of MC3T3-E1 cells, but the coated alloy showed better performance and obviously promoted the differentiation of the osteoblasts.

CONCLUSIONAlloying between Ta and Ti can be accomplished successfully by laser melting technique, and the alloy obtained has ideal surface structure and good biocompatibility.

3T3 Cells ; Alloys ; Animals ; Biocompatible Materials ; Cell Differentiation ; Cell Proliferation ; Materials Testing ; Mice ; Microscopy, Electron, Scanning ; Osteoblasts ; cytology ; Porosity ; Tantalum ; Titanium