Serious troubles were caused by the highly heterogeneous cancer cells in clinical cancer treatment.In recent years,molecular characterization of tumors and corresponding individualized cancer management has become hotspots for cancer research.Circulating tumor cells (CTCs) are shed from primary solid tumors or metastases into the peripheral blood.CTCs could serve as liquid biopsy for patients with cancer,with non-invasive,real-time and repeatable access.Serial monitoring of the amount and molecular characteristics of CTCs in the blood samples has significant clinical value for prognostic prediction,targeted drugs choice and real-time evaluation of clinical effectiveness in individualized cancer treatment.This review summarizes current methods for the CTC isolation and detection,and discusses the perspectives of CTC analyses in real-time personalized cancer therapy.Some future research directions in this field are proposed.

Circulating tumor cells are tumor cells that spontaneously or, following operations, migrate into the peripheral blood circulation from the primary tumor or metastatic tumor. As diagnostic, prognostic and predictive biomarkers in many types of cancers including breast cancer, lung cancer, pancreatic cancer and colorectal cancer, circulating tumor cells contribute to the early diagnosis of cancers, drug resistance monitoring, prognostic assessment, survival analysis, detection of tumor recurrence and evaluation of drug efficacy to assist in treatment decision?making and adjustment of treatment plans. However, the current approaches to the detection of circulating tumor cells still have limitations, and the development of new detection methods with higher sensitivity and specificity will be helpful for better use of these cells. Herein the authors review the research progress in circulating tumor cells in terms of the detection techniques, clinical applications of circulating tumor cells, and their prospects in future clinical practice.

OBJECTIVETo observe the clinical efficacy of Penyanqing Capsule (PYQC) in treating pelvic inflammation of Qi-stagnation with blood stasis syndrome.

METHODSThe randomized, single blinded, parallel positive drug controlled method was adopted, with 82 patients assigned into two groups by envelop method. The 42 patients in the treated group received PYQC 3 times a day, 4 capsules each time taken orally; the 40 patients in the control group were given orally Fuyankang tablets (FYKT) 3 times a day, 6 tablets each time. The therapeutic course for both groups was 2 months, and 2 courses of treatment were given successively to observe the comprehensive effect, changes of symptoms and signs before and after treatment. The effects of PYQC on hemorrheological character in part of the patients and on the pathogenetic chlamydia and mycoplasma were also observed.

RESULTSThe total effective rate in the treated group was 83.3%, which was insignificantly different from that in the control group (77.5%, P > 0.05). However, PYQC could significantly lower the hemorrheologic indexes in patients and showed definite influence on the pathogenetic chlamydia and mycoplasma.

CONCLUSIONPYQC has good therapeutic effect in treating chronic pelvic inflammation of Qi-stagnation with blood stasis syndrome, and showed definite effect on chlamydia and mycoplasma.

Adolescent ; Adult ; Blood Circulation ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Pelvic Inflammatory Disease ; drug therapy ; Qi ; Single-Blind Method

Objective:To assess the effect of serum heat inactivation on the detection of 2019 novel coronavirus (2019-nCoV) specific IgM and IgG antibodies by colloidal gold method.Methods:The serum specimens were collected from a total of 106 Coronavirus Disease 2019 (COVID-19) patients and 52 control subjects. Both the fresh serum and the heat inactivated serum samples from the same patient were detected simultaneously with the 2019-nCoV IgM and IgG antibodies detection kit (colloidal gold method). According to the patient′s onset time, the positive rates of antibodies production profile were calculated. The influence of heat inactivation on the detection rates of antibodies at different stages of disease after onset was analyzed.Results:The test results of the specimens of the healthy control group before and after inactivation were all negative. For the 106 specimens of COVID-19 patients, the detection rates of 2019-nCoV specific IgM and IgG antibodies were reduced after heating at 56 ℃ for 30 min. The positive rates of IgM antibodies significantly decreased from 66.04% (70/106) to 43.40% (46/106) ( χ2=22.042, P=0.000), while the positive rates of IgG antibodies slightly decreased from 81.13% (86/106) to 76.42% (81/106) ( χ2=0.800, P=0.063). Further analysis revealed that there was a significant difference in the positive rates of IgM antibodies before and after heat inactivation in the 3rd, 5th and 6th week after onset. However, there was no statistically significant difference in the detection rates of IgG antibodies before and after serum heat inactivation in different periods of onset. Conclusions:Heat inactivation significantly decreased the detection rates of 2019-nCoV specific IgM antibodies, which may lead to serological false negative results.

Objective:This study aims to evaluate the performance of digital PCR (dPCR) detecting multiple and single copies genes of the Epstein-Barr virus (EBV) for nucleic acid quantification and explore their applicability in clinical settings.Methods:Compared the sensitivity, specificity, precision, lower limit of detection (LoD), and linearity for multicopy BamHI-W dPCR and single-copy EBNA1 dPCR systems. Linear regression analysis using the least squares method was employed to evaluate the linearity. Additionally, we analyzed plasma samples from 182 patients with suspected EBV-related diseases between January and July 2022 at the Southern Medical University Southern Hospital, using both dPCR and quantitative PCR (qPCR) for EBV DNA quantification. Linear regression analysis using the least squares method was conducted to assess their quantitative correlation.Results:The dPCR systems for both multicopy and single-copy genes showed excellent linearity ( R 2 values of 0.992 and 0.997, respectively, both P<0.001). The LoD were 188 IU/ml for BamHI-W gene and 358 IU/ml for EBNA1 gene dPCR systems. The logarithmic coefficient of variation ( CV) values for high-concentration samples (1 000 000 IU/ml) were 0.34% and 0.21% for the BamHI-W gene and EBNA1 gene dPCR assays, respectively, while for low-concentration samples (5 000 IU/ml) were 0.98% and 0.64%, respectively. In the detection of seven common clinical infectious pathogens and EBV positive samples, only EBV-positive samples yielded positive signals in the dPCR detection system, with no cross-reaction with other pathogens. In 182 samples, the positive detection rates were 47.80% (87/182) for BamHI-W gene and 35.16% (64/182) for EBNA1 gene dPCR, compared to 43.41% (79/182) for qPCR. Linear correlation analysis with qPCR showed R2 values of 0.837 for BamHI-W gene and 0.763 for EBNA1 gene dPCR (both P<0.001). The BamHI-W gene copy number ranged from 3 to 18 copies per clinical sample, with patient-specific variations. There was a high consistency in viral load trends between the multicopy BamHI-W gene and single-copy EBNA1 gene dPCR systems within individual patients. Conclusions:The dPCR methods detecting EBV multiple and single copies genes showed high sensitivity, specificity, precision, and quantitative accuracy, suitable for clinical sample analysis. The multicopy BamHI-W gene dPCR method notably enhances detection sensitivity and can be used as a supplement to current EBV DNA load detection methods, especially in low-concentration samples. For within-patient EBV DNA monitoring, the multicopy gene method proves more effective, while inter-patient comparisons might necessitate single-copy gene methods or normalize them using the same standard.

Objective: To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (CYP2C9), CYP2C19, CYP4F2, Vitamin K epoxide reductase (VKORC1) and ATP-binding cassette subfamily B member1 (ABCB1) gene, which were associated with warfarin and clopidogrel, based on liquid phase chip technology. Methods: Method establishment. The eight gene sequences near targeted sites related to warfarin and clopidogrel were found in Genbank, and the specific primers and probes were designed. Through multiple PCR amplification, followed by allele specific primer extension (ASPE), and MagPlex-Tag microspheres hybridization, the suspension array Luminex 200 system step-by-step, the genotypes were determined by fluorescence signal. The reaction system was optimized and its methodological evaluation was performed. 260 patients with antithrombotic therapy from Dongguan houjie hospital were recruited in this study form June 2017 to December 2018. The eight genotypes of the 260 patients were detected by the established method, and the results were compared with the sequencing results. Results: The results of 260 samples showed that allelic median fluorescence intensity (MFI) ratios of homozygotes (mutant/wild-type) were all greater than 0.9 or less than 0.1, and all the allelic MFI ratios of heterozygotes were between 0.3 and 0.6. The within run and between run coefficients of variance for allelic MFI ratios were lower than 6.4% and 10.9%, respectively. The minimum DNA template requirements was 0.75ng. The genotypes of 260 patients determined by the established method were completely concordant with the sequencing results. Conclusion A method was established successfully for rapid detecting the genotypes which associated with warfarin and clopidogrel based on liquid phase chip technology.