OBJECTIVETo investigate the biotransformation of daphnetin by suspension transgenic hairy root of Polygonum multiflorum and provide a biotechnological method for large-scale production of the daphnetin-8-O-beta-D-glucoside using this new culture system.

METHODDaphnetin was added into the media of suspension to culture 36 h. The biotransformation product was detected with TLC and HPLC, and isolated by various chromatographic methods. The influence of co-cultured time on conversion ratio, content of degradation product and the reason for the degradation of product II were investigated using HPLC.

RESULTOne biotransformation product, daphnetin-8-O-beta-D-glucoside, was obtained, the optimal co-cultured time in suspension hairy root of P. multiflorum was 36 h with the highest biotransformation molar ratio of 32.11%, the sucrose medium (sucrose-only) can increase the biotransformation molar ratio to 72.44%. The result demonstrated that the degradation products of the product II was induced by the MS medium.

CONCLUSIONThe potential application of suspension transgenic hairy root of P. multflorum in the sucrose-only medium on generating daphnetin-8-Obeta-D- glucoside could be prospective.

Biotransformation ; Coculture Techniques ; Glucosides ; metabolism ; Plant Roots ; genetics ; metabolism ; Plants, Genetically Modified ; Polygonum ; genetics ; metabolism ; Umbelliferones ; metabolism

Objective To compare the efficacy of uniform multi-drug therapy (UMDT) versus routine multi-drug therapy (RMDT) for the treatment of multi-bacillary (MB) leprosy patients based on bacterial index changes and frequencies of leprosy reaction.Methods This study recruited newly diagnosed leprosy patients after taking informed consent in three districts of Guizhou province as well as in one district of Yunnan province from November 2003 to June 2005.The patients received 6-month UMDT or 2-year RMDT.Clinical follow up and bacterial reexamination were carried out once a year.Changes of bacterial index (BI) and frequencies of leprosy reaction were compared between the patients receiving RMDT and UMDT.Results A total of 166 patients received UMDT and 170 received RMDT in this study.Among the UMDT-treated patients,114 were skin smear positive,and 83 had been followed up for 42 months; of the RMDT-treated patients,149 underwent all the bacterial examinations during a 48-month follow up.The mean bacterial index decreased from 2.84 before treatment to 0.33 at the end of the 42-month follow up in the 83 patients,and from 2.55 to 0.26 at the end of the 48-month follow up in the 149 patients,with no significant difference in the changes of bacterial index between the two groups (t =0.77,P ＞ 0.05).Bacterial index became negative in 73.5％ (61/83) of the UMDT-treated patients and in 77.2％ (115/149) of the RMDT-treated patients (x2 =0.40,P＞ 0.05)at the end of follow up.During the follow up peroid,the incidence of type Ⅰ leprosy reaction was 14.6％ (13/89) in the UMDT group,significantly higher than that in the RMDT group (3.4％ (5/149),x2 =10.08,P＜ 0.01 ).Conclusions There is no significant difference in mean bacterial index changes and bacterial clearance rate during the follow up peroid between UMDT- and RMDT-treated patients.The incidence of type Ⅰ leprosy reaction is higher in the UMDT group than in the RMDT group,and further investigation is needed to clarify the mechanisms underlying the phenomenon.

Objective To detect gene mutations associated with dapsone-,rifampicin-and ofloxacinresistance in lesions of patients with recurring or treatment-resistant leprosy collected from 2010 to 2011.Methods Clinical data and lesional specimens were collected during 2010-2011 from patients with recurring or treatment-resistant leprosy who were diagnosed and reported by provincial centers for leprosy control.Mycobacterium leprae DNA was extracted from the specimens and subjected to PCR for the amplification of folP1,rpoB and gyrA genes.The PCR products were directly sequenced and BLAST program was used to compare the sequence of isolated strains with the reference sequence in GenBank.Results Twenty-four patients were enrolled in this study,including 13 with recurring leprosy and 11 with treatment-resistant leprosy.Twenty-one patients showed positive PCR results in all the three regions.Of these PCR-positive specimens,3 from 1 patient with recurring and 2 patients with resistant leprosy harbored a point mutation,acc (threonine)→gcc (alanine),at codon 53 in the floP1 gene,1 from a patient with recurring leprosy harbored a missense mutation,gat (aspartic acid) → aac (asparagine),at codon 441 in the rpoB gene.Conclusions Mutations are detected in the folP1 and rpoB genes,which are associated with the resistance to dapsone and rifampicin respectively,but not in the ofloxacin resistance-associated gyrA gene,in Mycobacterium leprae isolates from patients with recurring or treatment-resistant leprosy.

OBJECTIVETo investigate the influence of 2,3-dihydrocyclopenta coumarins to the growth of transgenic hairy root of Polygonum multiflorum and the accumulation of active ingredient stilbene glucoside in the culture.

METHODIn different culture time, 2,3-dihydrocyclopenta coumarin compounds were administrated to the hairy roots. The yield of stilbene glucoside and the growth of the hairy roots were determined after some period of co-culture. Through the investigation of the different concentrations of the 2,3-dihydrocyclopentacoumarin, and sub-culture times, the optimal experimental conditions were established.

RESULTThe optimal administration time of the 2,3-dihydrocyclopentacoumarin was on the fourth day, the optimal concentration of 2,3-dihydrocyclopentacoumarin was 0.025 g x L(-1). Under this condition, the growth of the hairy root greatly increased. Comparing with control group, the accumulation of stilbene glucoside was increased about four times.

CONCLUSION2, 3-dihydrocyclopentacoumarins could promote the hairy hair root growth and stimulate the stilbene glucoside accumulation.

Coumarins ; pharmacology ; Glucosides ; metabolism ; Plant Roots ; drug effects ; growth & development ; metabolism ; Polygonum ; drug effects ; growth & development ; metabolism ; Stilbenes ; metabolism

Objective:To access the feasibility and efficacy of percutaneous endoscopic debridement (PED) combined with percutaneous pedicle screw fixation (PPSF) in the treatment of lumbar pyogenic spondylodiscitis.Methods:49 patients (male 29, female 20), aged 51.2±13.9 years (range 19-81 years), who were diagnosed with lumbar pyogenic spondylodiscitis and received PED with PPSF in Orthopedic Department, Sun Yat-Sen Memorial Hospital and Zhuhai People's Hospital from January 2014 to March 2017, were retrospectively reviewed. The patients were operated in the prone position with the infected locus thoroughly debrided, vertebrae fixed and clinical outcomes were assessed by observing the changes of complaining symptoms, laboratory parameters, clinical functional scores (American Spinal Injury Association impairment scale, AIS; visual analog scale, VAS; Oswestry disability index, ODI) and imaging studies during perioperative and follow-up stages.Results:The mean operative time was 110.1±19.8 min (80-165 min), with intra-operative blood loss 47.8±20.6 ml (range 20-120 ml). All patients reported relief of back pain. Causative pathogens were identified in 36 of 49 biopsy specimens, with staphylococcal bacteria being the most prevalent strain (accounting for 50.0%). During 3-12 months' follow-up, 95.9% (47/49) patients' infection was well-controlled. At 3 month post-operative, C-reactive protein declined from 62.1±37.2 mg/L to 7.5±5.8 mg/L, white blood cell declined from (14.2±3.9)×10 9/L to (6.2±1.1)×10 9/L, ESR declined from 90.3±37.4 mm/1 h to 16.9±7.2 mm/1 h, and the values at 3 months post-operative had significant difference compared with values at pre-operative ( t=10.15, P<0.001; t=13.49, P<0.001; t=13.82, P<0.001). Spontaneous fusion was observed among 56.8% (21/37) of the patients during long-term radiological follow-ups (more than 1.5 years). At the last follow-up, the VAS declined from 7.4±0.6 points pre-operative to 0.5±0.3 post-operative, ODI declined from 78.2%±9.1% pre-operative to 14.0%±8.6% post-operative, and the values at the last follow-up had significant difference compared with values at pre-operative ( t=72.00, P<0.001; t=35.89, P<0.001). There were 38 cases of AIS E, and 11 cases of AIS D at pre-operative, while 43 cases of AIS E and 6 cases of AIS D. However, there were 11 patients developed post-operative complications, among whom 2 with recurrent infection, 2 with secondary neurological impairment. Conclusion:PED combined with PPSF effectively eliminated infected locus, stabilized the affected vertebrae, improved patients' clinical outcomes with small trauma, thereby offering an alternative for the treatment of lumbar pyogenic spondylodiscitis.

By analyzing and comparing the IVD product classification supervision system in the EU's new IVDR, old IVDD and related guidance documents, concepts of related regulatory reforms were analyzed and explored, to provide references for industrial personnel to understand and master the new EU IVD classification system, and for China's ongoing classification management reform.
European Union ; Industry ; Reagent Kits, Diagnostic

Objective To evaluate the changes in expression of cold-inducible RNA-binding protein (CIRP) in hippocampus during brain injury in a rat model of cardiac arrest-cardiopulmonary resuscitation.Methods Seventv-two clean-grade healthy male Sprague-Dawley rats,weighing 280-350 g,aged 8-10 weeks,were divided into 2 groups using a random number table:sham operation group (S group,n=18) and ischemia-reperfusion group (I/R group,n=54).Tracheal intubation was performed and artery and veins were punctured in group S.Ventricular fibrillation was induced by transoesophageal cardiac pacing to establish the model of cardiac arrest in group I/R.Rats were sacrificed at 12,24 and 48 h after resuscitation and the hippocampus was harvested for determination of CIRP,tumor necrosis factor-alpha (TNF-α) and interleukin-lbeta (IL-1β) protein and mRNA expression (by quantitative polymerase chain reaction or Western blot) and for determination of pathological changes of hippocampi (with a light microscope).Results Compared with group S,the expression of CIRP mRNA in hippocampus was up-regulated at 24 and 48 h after resuscitation,the expression of TNF-α mRNA was up-regulated at 12,24 and 48 h after resuscitation,the expression of IL-1β mRNA was up-regulated at 12 and 24 h after resuscitation,and the expression of CIRP,TNF-α and IL-1β was up-regulated at 12,24 and 48 h after resuscitation in group I/R (P＜0.05).Pathological changes in hippocampal CA1 region were found in group I/R.Conclusion The expression of CIRP in hippocampus is up-regulated,which promotes central inflammatory responses during brain injury in a rat model of cardiac arrest-cardiopulmonary resuscitation.

OBJECTIVE: Discuss the working ideas of the dynamic adjustment mechanism of medical device classification in the United States, and provide reference for the construction of medical device related mechanisms in China. METHODS: Collect and interpret the documents of regulatory background, procedures and orders of the dynamic adjustment mechanism of the medical device classification in the United States, and summarize the overall situation and specific cases of the medical device classification adjustment under this mechanism in recent years. RESULTS: The US work idea of the medical device classification dynamic adjustment mechanism is based on the latest valid scientific evidence, conducting risk analysis and identification, and determining the corresponding measures. CONCLUSIONS During the adjustment process, industry stakeholders have repeatedly discussed and achieved final agreement. Its procedures and working ideas can be used as a reference for China's work.
China ; United States ; United States Food and Drug Administration

By comparing new medical device regulations with existing directives in the European Union, the revising ideas of new EU medical device regulations, especially the changes of the classification management concepts, were analyzed to provide clues for the industry to interpret the new EU regulations, and references for the classification management innovation in our country.
Equipment and Supplies ; classification ; European Union ; Government Regulation ; Industry

Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.