Objective To explore the application value of automatic nucleic acid extractor combined with vacuum concentrator in forensic DNA extraction. Methods Gradient samples of human peripheral venous blood were collected at 40, 80, 120, 160, 200, 240, 280 and 320 fold dilution. The samples of each gradient were treated with no inhibitor, black oil, rust, fruit acid, tin foil and indigo, respectively. The automatic nucleic acid extractor was used for DNA purification and extraction of the above samples. The extracted DNA eluent （6 μL） was taken for amplification directly, and the rest was concentrated by vacuum concentrator. DNA was amplified and examined using the Investigator 26plex QS kit before and after concentration. Results Only gradient samples treated with fruit acid obtained complete STR typing results at 40 fold dilution. The other 5 methods obtained complete STR typing results at 40-160 fold dilution. The results of STR typing after DNA concentration showed that the average peak height and detection rates of gene loci both increased to a certain extent, but the effect was not obvious. Conclusion The automatic nucleic acid extractor has an efficient inhibitor removal ability and high extracting efficiency of DNA. The vacuum concentrator can concentrate DNA samples to a certain extent. Combining the automatic nucleic acid extractor with the vacuum concentrator can improve the examination success rate of forensic materials.
DNA/genetics* ; DNA Fingerprinting ; Humans ; Microsatellite Repeats ; Nucleic Acids ; Vacuum

OBJECTIVETo classify colorectal carcinoma (CRC) by TNM staging integrated with the gene expression profile and copy number variation (CNV).

METHODSProfile data of gene expression and CNV of CRC were downloaded from public database and processed with batch bias adjustment, quartile normalization, missing value estimation and feature filtration. The processed profiles of mRNA and CNV were introduced into the codes of Bayesian consensus clustering (BCC) method and were used to calculate the subclasses of CRC. With the follow-up information of disease free survival of CRC patients, the prognostic values of the subclasses was investigated and the software of function enrichment analysis was employed to discover the major pathway signaling to each interesting subclass. All statistic analyses were performed under R-3.0.1 environment or by using SPSS 16.0 software.

RESULTSProfile data of gene expression and corresponding CNV from 335 CRC patients with TNM stage Ⅱ-Ⅲ and followed-up information were obtained. After feature filtering, the profiles contained 1578 probes of mRNA and 345 location of CNV. Four CRC subclasses were identified by the integrative analysis with BCC, and the concordances of BCC subclasses and each of gene-based subclasses (Cramer's V=0.49), CNV-based subclasses (Cramer's V=0.51) and Marisa's subclasses (Cramer's V=0.32) were statistically significant (Ps<0.001). Among BCC subclasses, BCC-I had a favorable prognosis, while BCC-Ⅳ had more unfavorable prognosis. The differences of prognosis were significant among BCC-I, BCC-(Ⅱ+Ⅲ) and BCC-Ⅳ with an overall log-rank P<0.001. The top enriched function was DNA damage and repair signaling when BCC-I compared to BCC-Ⅳ, and the new subgroups classified by the genes associated with enriched signaling had the better prognostic value than BCC subclasses but both of them were significantly correlated (Cramer's V=0.39, P<0.001).

CONCLUSIONBCC method is effective to integrate multi-type genomic data for molecular classification of colorectal carcinoma, and the BCC-Ⅳ subclass has poor prognosis, which may be associated with the decreased repairing function of DNA damage.

Colorectal Neoplasms ; classification ; genetics ; pathology ; DNA Copy Number Variations ; Gene Expression Profiling ; Humans ; Neoplasm Recurrence, Local ; Postoperative Period ; Prognosis ; Transcriptome

Objective:To summarize the clinical application value of color Doppler ultrasound (CDU) in the nursing observation of flap vascular circulation after free flap transplantation.Methods:From September 2018 to November 2020, 18 patients with skin defect of limbs treated by free flap transplantation in the Second Affiliated Hospital of Wenzhou Medical University were monitored by skin color, skin temperature, skin tension, capillary reaction time and CDU. When the vascular circulation disorder was found, the patients should be treated with volume expansion, rehydration, local massage of the flap, anticoagulation and microcirculation improvement. Positive drainage under the flap to prevent infection, combined with hyperbaric oxygen treatment, accurate and timely exploration of vascular crisis to avoid flap necrosis.Results:Postoperatively, CDU monitoring found that 2 cases had regional vascular circulation disorder, 1 case had complete embolization of the main vessels and perforators in the flap, the flap was thinned and changed to full thickness skin grafting, the deep tissue exposed area was healed by dressing change, and 1 case had incomplete embolization of the flap′s reflowing vein. After operation, the venous crisis was relieved and the flap survived completely.Conclusions:Comprehensive observation is needed after free flap transplantation. Combined with four routine vascular circulation indexes, CDU monitoring of flap blood flow can provide more objective, accurate and detailed basis for vascular circulation status, and can accurately find the vascular circulation disorder after free flap transplantation in early stage, and understand the vascular flow status of the flap, so as to effectively avoid the continuous progress of vascular crisis, to avoid flap necrosis and improve the survival rate of flap.

Objective:To construct a teamwork model, Partnership Rehabilitation Therapy (PRT), for therapists in critical wards for patients with Corona Virus Disease 2019 (COVID-19), and observe its effect. Methods:PRT had been developed, in which one therapist (main) implementing therapy and another (assistant) monitoring and supporting in the treatment. Eleven COVID-19 patients from infectious critical ward were treated with PRT. The behavior safety of therapists was recorded during the treatment, and the patients were assessed with Borg Index, Cough Score, Miller Sputum Grading and World Health Organization Disability Assessment Schedule (WHODAS) 2.0 before and after treatment. Results:No physiotherapist was infected by COVID-19. Seven times of infection risks were recorded and avoided, and six times of treatment risks were corrected instantly. All the patients improved in Borg Index (P < 0.01), Cough Score (P < 0.05), Miller Sputum Grading (P = 0.02) and WHODAS 2.0 (P < 0.01) after a 1-week physical therapy. Conclusion:Based on the Family International Classifications, a teamwork model is established, which provides a safe and practicable way for rehabilitation for COVID-19 patients in critical wards.