Twelve pre-processing protocols for formalin-fixed paraffin-embedded(FFPE)tissue samples were developed by orthogonal experimental design,incorporating different dewaxing buffers(Triton X-100 and xylene),lysis buffers(TFE and RapiGest),and enzyme digestion methods(iST,SP3,and FASP)to explore the optimal experimental conditions.These protocols were assessed based on protein and peptide identification depth,identification stability,and quantitative levels of protein abundance.The results indicated that Triton X-100 and xylene minimally impacted proteomics identification,whereas the TFE lysis buffer and iST digestion method significantly enhanced the proteomics analysis of FFPE samples.Considering the potential toxicity of xylene,the TTI protocol based on Triton X-100,TFE,and iST was determined to be the optimal choice.This protocol exhibited the best repeatability and stability,and a higher number of proteins associated with significant biological functions were identified.In conclusion,the established TTI protocol offered an efficient and comprehensive approach for proteomic analysis of FFPE samples,significantly enhancing the repeatability and stability of protein identification.

According to the pathological characteristics of symptomatic chronic thoracic and lumbar osteoporotic vertebral fracture (SCOVF), the different clinical treatment methods are selected, including vertebral augmentation, anterior-posterior fixation and fusion, posterior decompression fixation and fusion, and posterior correction osteotomy. However, there is still a lack of a unified understanding on how to choose appropriate treatment method for SCOVF. In order to reflect the new treatment concept and the evidence-based medicine progress of SCOVF in a timely manner and standardize its treatment, the clinical guideline for surgical treatment of SCOVF is formulated in compliance with the principle of scientificity, practicability and advancement and based on the level of evidence-based medicine.

Hyoscyamine and scopolamine are two main alkaloids in Atropa belladonna with great medicinal value. In this paper, the contents of hyoscyamine and scopolamine, the upstream products in alkaloid synthesis, and the expression levels of key enzyme genes PMT, TRⅠ and H6H in secondary metabolism of A. belladonna seedlings were measured to clarify the mechanism of nitrogen forms regulating alkaloids synthesis.The results showed that the 50/50 (NH⁺₄/NO⁻₃) treatment was more favorable for the accumulation of alkaloids and the conversion of hyoscyamine to scopolamine. The content of putrescine was almost consistent with the change of key enzymes activities in the synthesis of putrescine, they both increased with the rise of ammonium ratio, reaching the highest at 75/25 (NH⁺₄/NO⁻₃). The detection of signaling molecule nitric oxide (NO) showed that the NO concentration decreased with the decrease of nitrate proportion. Further detection of gene expression levels of PMT, TRⅠ and H6H in TAs synthesis pathway showed that a certain amount of ammonium promoted the expression of PMT and H6H in roots. When the ratio of ammonium to nitrate was 50/50, PMT, TRⅠ and H6H in leaves and roots had higher expression levels. It can be speculated that the regulation of the formation of hyoscyamine to scopolamine by nitrogen forms mainly through affecting the expression of key enzyme genes. 50/50 (NH⁺₄/NO⁻₃) treatment increased the gene expression of TRⅠ in both leaves and roots as well as PMT and H6H in roots, promoting the synthesis of putrescine to hyoscyamine and the conversion of hyoscyamine to scopolamine.
Atropa belladonna ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Hyoscyamine ; biosynthesis ; Mixed Function Oxygenases ; Nitrogen ; metabolism ; Scopolamine ; metabolism

AIM To establish an HPLC method for the simultaneous content determination of four constituents in Hydnocarpus anthelmintica Pierre.METHODS The analysis of 60％ ethanol extract from H.anthelmintica was carried out on a 35 ℃ thermostatic Diamonsil C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of 80％ acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 347 nm.RESULTS Luteolin,hydnocarpusol,chrysoeriol and hydnocarpin showed good linear relationships within the ranges of 0.0523 6-1.047 μg (r =0.999 9),0.011 24-0.224 8 μg (r =0.999 9),0.029 46-0.589 2 μg (r =0.999 9) and 0.130 5-2.609 μg (r =0.999 9),whose average recoveries were 102.45％ (RSD=1.9％),98.66％ (RSD=1.8％),97.60％ (RSD=1.6％) and 97.88％ (RSD=1.4％),respectively.CONCLUSION This simple,accurate and sensitive method can be used for the quality control of H.anthelmintica.

Objective To investigate the prevalence of periodontal pathogens from saliva of patients with type 2 diabetes mellitus (T2DM),and to characterize the association between the glucose status and periodontal pathogens in oral cavity.Methods All the subjects were hypertension patients under regular care at Beijing hypertension prevention and management institute.Whole unstimulated saliva samples were collected from 45 non-diabetic subjects (non-DM group),80 well-controlled diabetic patients (DM-well group) and 100 poor-controlled diabetic patients (DM-poor group).DNA was extracted from the salivary deposition,Porphyromonas gingivalis (Pg),Tannerella forsythia (Tf) and Treponema denticola (Td) were detected by polymerase chain reaction (PCR) method based on 16SrRNA.Prevalence and quantity of the pathogens under different glucose states were compared and logistic regression model was set to analyze the factors related to each bacterium.Results The prevalence of Tf in DM-well group and DM-poor group was significantly lower than that of non-DM group [81％ (65/80),80％ (80/100) vs 91％ (41/45),P =0.048],meanwhile the quantity of Tf was also lower than that of non-DM group[1.9 (2.6),2.1(5.3) vs 3.4 (6.4)] (P ＞ 0.05).With the worsening of glucose control,the quantity of Tf was declining (P =0.032).However,the prevalence and the quantity of Pg,Td in 3 groups had no statistical differences(P ＞0.05).After adjusting age,gender,number of missing teeth and other periodontal parameters,OR of having Tf in saliva from DM-well group and DM-poor group was 0.58 and 0.53,respectively.Conclusions Abnormal blood glucose state may affect the colonization of Tf in oral cavity.

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome