Objective:Prepared the specific chicken egg yolk immunoglobulins (IgY) against the cell wall protein of Trichophyton mentagrophytes (tmCWP) and detected its biological activities,which was to establish the basis for the preventment and treatment in dermatophytes disease.Methods: In this work,tmCWP was extracted and purified by cold alkali method,and being used as immunogen to immunized healthy laying hens.The IgY was extracted from the egg yolk by polyethylene glycol method and purified by saturated ammonium sulfate method,respectively.The concentration of the extracted IgY was detected by Bradford method.The purity and molecular weight of the specific anti-tmCWP IgY were analysed by SDS-PAGE.The titer of IgY was obtained by ELISA.The immunoreactivity of IgY was performed by Western blot.Results: The purity of the extracted IgY reached to 87.27％.ELISA indicated that the titer of the specific anti-tmCWP IgY gradual rised 20 days after primary immunization and reached to the highest value (1∶32 000) after 45 days.Western blot revealed that the specific IgY showed a good immunoreactivity and a specifically combination capacity.Conclusion: In our work,the tmCWP could be regarded as the immunogen to prepare the specific anti-tmCWP IgY,which could provide a novel thought for the therapy of Trichophyton mentagrophytes infection.

Objective To investigate the mechanism of radiosensitization by cetuximab (C225) on human tongue cancer Tca8113 cell line in vitro.Methods Tca8113 cell line with and without C225 treatment received 6 MV X-ray irradiation of different doses (0, 2,4,6, 8 and 10 Gy). Cell proliferation,cell-cycle distribution and clonogenic survival were analyzed through cell counting,MTT,colony formation assay,and flow cytometry,respectively.Results After irradiation of different doses,the growth inhibition rates in C225 group were higher than control (t =- 15.6 - -3.0,P＜0.05),the radiobiological parameters (D0,Dq,N,and SF2 ) in C225 group were lower than control so that SER of C225 group was 1.353,and the proportions of G0/G1 cells in C225 group were higher than control ( t =-7.64,-7.89,-4.78,P ＜0.05 ) at 4,6,8 Gy.When the irradiation doses increased,the early phase apoptosis in both groups increased at first and then decreased with the maximum difference at 4 Gy [(7.96±0.36)％ in C225 group and (4.13 ±0.29)％ in control group,t =-12.75,P＜0.01 ].Conclusions C225 has radiosensitization effect on Tca8113 cell line,possible through Go/G1 arrest and induction of apoptosia.

Objective To evaluate the prognosis-related factors of colorectal cancer patients with positive PD-L1 expression in liver metastases after hepatectomy. Methods We reviewed retrospectively the clinical data of 68 colorectal cancer patients with positive PD-L1 expression in liver metastases receiving personalized comprehensive treatment which was mainly consisted of surgical resection. We observed the results and prognosis after surgical resection and analyzed related factors. Results Univariate analysis showed that no radiotherapy, N stage, RAS mutation status, T stage, dMMR, Duck stage, disease free interval from primary to metastases≤12 months and largest hepatic tumor diameter > 5 cm had obvious significance (all P < 0.05). Multivariate logistic analysis regression revealed that without dMMR (P=0.012), Duck A stage (P=0.000), disease free interval from primary to metastases > 12 months (P=0.020) and largest hepatic tumor diameter < 5 cm (P=0.006) were independent protective factors for the prognosis of colorectal cancer patients with positive PD-L1 expression in liver metastases after surgical resection. Conclusion Personalized comprehensive treatment which is mainly consisted of surgical resection still has a good effect on colorectal cancer patients with positive PD-L1 expression in liver metastases after hepatectomy. Without dMMR, Duck A stage, disease free interval from primary to metastases > 12 months and largest hepatic tumor diameter ≤5cm are independent protective factors for patients' prognosis.

Background 5-hydroxytryptamine 2A receptor (5-HT2A) gene has been regarded as a candidate gene for susceptibility to schizophrenia. In particular, the 5-HT2A receptor has received much attention because it demonstrates to be an important site of action of atypical antipsychotic agents to alleviate negative symptoms. The current study investigated whether the 5-hydroxytryptamine 2A receptor (5-HT2A) gene T102C polymorphism was associated with treatment response to risperidone in the first episode Chinese patients with schizophrenia.Methods 201 first episode Chinese Han patients with schizophrenia were given risperidone for up to 56 days.Genotyping of 5-HT2A gene T102C polymorphism were performed by using PCR-RFLP. The Positive and Negative Symptom Scale (PANSS) was used for the evaluation of the severity of psychotic symptoms before and after 8 weeks treatment with risperidone.Results 5-HT2A receptor 102 -T/T genotype was significantly associated with both the PANSS total and negative syndrome subscale scores before treatment, and with the reduction rates in both the PANSS total and negative syndrome subscale scores after eight weeks risperidone treatment.Conclusions The results suggest that 5-HT2A T102C A/A1 genotype subgroup influences individual response to risperidone in first-episode Chinese patients with schizophrenia.

AIM: To construct pVAX1-GrB. METHODS: Lymphocytes from human laryngeal carcinoma tissue were separated from tumor tissue. The fragment of granzyme B (GrB) was amplified by RT-PCR and was recombined to the downstream of T7 promoter in the vector pVAX1. The construction was transfected into Hep2 cells with lipofectamine 2000. The expression of protein was identified by indirect immunofluorescent antibody assay. RESULTS: It has been proved that the sequence of the RT-PCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The GrB cDNA fragment was cloned into the vector of pVAX1 in the right direction and the open reading fragment of GrB was maintained. The target protein was detected in the transfected Hep2 cells. CONCLUSION: The pVAX1-GrB plasmid was successfully constructed and expressed. [

No abstract available.
Bone Marrow* ; Flow Cytometry* ; Myelodysplastic Syndromes*

To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chro-matography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work il-lustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.At-tributes of nanobody based pharmaceutical assays are discussed.

Developing analytical methods for the chemical components of natural medicines remains a challenge due to its diversity and complexity. Miao-Fu-Zhi-Tong (MFZT) granules, an ethnic Yi herbal prescription, comprises 10 herbs and has been clinically applied for gouty arthritis (GA) therapy. Herein, a series of chemical profiling strategies including in-house library matching, molecular networking and MS/MS fragmentation behavior validation based on ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) were developed for qualitative analysis of MFZT granules. A total of 207 compounds were identified or characterized in which several rare guanidines were discovered and profiled into alkyl substituted or cyclic subtypes. Moreover, network pharmacology analysis indicated that MFZT's anti-gout mechanism was mostly associated with the nuclear factor kappa-B (NF-κB) signaling, nucleotide oligomerization domain (NOD)-like signaling and rheumatoid arthritis pathways, along with the synergistic effect of 84 potential active compounds. In addition, a quantitative analytical method was developed to simultaneously determine the 29 potential effective components. Among them, berberine, pellodendrine, 3-feruloylquinic acid, neoastilbin, isoacteoside and chlorogenic acid derivatives at higher concentrations were considered as the chemical markers for quality control. These findings provide a holistic chemical basis for MFZT granules and will support the development of effective analytical methods for the herbal formulas of natural medicines.
Humans ; Chromatography, High Pressure Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Drugs, Chinese Herbal/chemistry* ; Quality Control ; Arthritis, Gouty