BACKGROUND: Microsatellite instability (MSI) is a key biomarker for cancer immunotherapy and prognosis. Integration of MSI testing into a next-generation-sequencing (NGS) panel could save tissue sample, reduce turn-around time and cost, and provide MSI status and comprehensive genomic profiling in single test. We aimed to develop an MSI calling model to detect MSI status along with the NGS panel-based profiling test using tumor-only samples. METHODS: From January 2019 to December 2020, a total of 174 colorectal cancer (CRC) patients were enrolled, including 31 MSI-high (MSI-H) and 143 microsatellite stability (MSS) cases. Among them, 56 paired tumor and normal samples (10 MSI-H and 46 MSS) were used for modeling, and another 118 tumor-only samples were used for validation. MSI polymerase chain reaction (MSI-PCR) was performed as the gold standard. A baseline was built for the selected microsatellite loci using the NGS data of 56 normal blood samples. An MSI detection model was constructed by analyzing the NGS data of tissue samples. The performance of the model was compared with the results of MSI-PCR. RESULTS: We first intersected the target genomic regions of the NGS panels used in this study to select common microsatellite loci. A total of 42 loci including 23 mononucleotide repeat sites and 19 longer repeat sites were candidates for modeling. As mononucleotide repeat sites are more sensitive and specific for detecting MSI status than sites with longer length motif and the mononucleotide repeat sites performed even better than the total sites, a model containing 23 mononucleotide repeat sites was constructed and named Colorectal Cancer Microsatellite Instability test (CRC-MSI). The model achieved 100% sensitivity and 100% specificity when compared with MSI-PCR in both training and validation sets. Furthermore, the CRC-MSI model was robust with the tumor content as low as 6%. In addition, 8 out of 10 MSI-H samples showed alternations in the four mismatch repair genes ( MLH1 , MSH2 , MSH6 , and PMS2 ). CONCLUSION MSI status can be accurately determined along the targeted NGS panels using only tumor samples. The performance of mononucleotide repeat sites surpasses loci with longer repeat motif in MSI calling.
Humans ; Microsatellite Instability ; Colorectal Neoplasms/diagnosis* ; Microsatellite Repeats/genetics* ; DNA Mismatch Repair

Objective : To investigate the epidemiological and clinical characteristics of cases with pertussis in Yiwu City, Zhejiang Province, so as to provide insights into pertussis diagnosis and control. Methods: A total of 186 patients with definitive diagnosis of pertussis in medical institutions in Yiwu City from November 1, 2020 to August 31, 2022 were recruited, and subjects' demographic, clinical symptoms and history of pertussis vaccination were collected using questionnaire surveys. The temporal, population, and spatial distribution and clinical symptoms of pertussis were analyzed using a descriptive epidemiological method, and the clinical characteristics of pertussis patients with different doses of pertussis vaccination were compared. Results: Pertussis was found to predominantly occur during the period between July and November (101 cases, 54.30%), and the three highest-incidence regions included Jiangdong Street, Beiyuan Street and Choujiang Street (87 cases, 46.77%). The 186 pertussis cases included 105 males (56.45%) and 81 females (43.55%), and included 144 cases with age of onset under 7 year (77.42%). Preschool and diaspora children were predominant among all pertussis cases, and the main clinical symptoms included spasmodic cough (97 cases, 52.15%), post-tussive vomiting (82 cases, 44.09%) and aggravated cough at night (77 cases, 41.40%). Routine blood tests measured 119 cases with abnormal white blood cell counts (63.98%), 137 cases with abnormal lymphocyte counts (73.66%), 39 cases with abnormal neutrophil counts (20.97%), 21 cases with abnormal platelet counts (11.29%) and 111 cases with abnormal hemoglobin concentrations (59.68%). There were 55 cases that were unvaccinated (29.57%), 23 cases that were not fully vaccinated (12.37%), and 108 cases that were fully vaccinated (58.06%). There were significant differences among pertussis cases with different doses of vaccination in terms of age, incidence of post-tussive vomiting, percentage of abnormal platelet counts and percentage of hemoglobin concentrations (all P<0.05). Conclusions The majority of pertussis cases are preschool and diaspora children in Yiwu City from November 1, 2020 to August 31, 2022, and the clinical symptoms mainly include spasmodic cough, post-tussive vomiting and aggravated cough at night, with atypical symptoms. The capability for differential diagnosis of pertussis is required to be improved in medical institutions.

Objective To investigate the effect of home diabetes care platform based on internet and family fixed partner on the continuous care of diabetes patients outside the hospital.Methods A total of 150 out-patients with diabetes were collected from June 2016 to November 2016,divided into family fixed partner group(group A),smart phone APP home diabetes care platform group(group B),family fixed partner combination with smart phone APP home diabetes care platform group(group C)with 50 cases each by random digits table method.The three groups received the same health education during their stay in hospital,patients in group A and group C were required to have family fixed partners,patients in group B and group C were required to receive the home diabetes care platform for smart phones APP after they left the hospital,the intervention time was six months,and the indexes of blood glucose metabolism,self-management ability of diabetes were assessed at the end of six months after intervention and before intervention.Results Fasting blood glucose and postprandial blood glucose and glycosylated hemoglobin values of the three groups after intervention were lower than those before intervention.The postprandial blood glucose and glycosylated hemoglobin values was(9.96±4.23)mmol/L,(7.16±1.47)%in group C,(13.78±3.34),(11.46±4.85)mmol/L and(8.46±2.21)%,(8.07±2.45)%in group A and B,the difference was significant(F=10.57,3.92,P＜0.05).The scores of self-management ability of diabetes of the three groups after intervention were all higher than those before intervention.The score of self-management ability of diabetes of item 1-6 was(6.45±1.65),(4.87±2.23),(6.17±2.12),(5.24±1.65),(4.67±2.13),(6.27±2.02)points in group C,(5.78±1.96),(3.63±2.14),(5.25±2.34),(4.12±1.97),(3.65±1.34),(5.26±2.21)points in group B,(5.04±1.78),(3.37±1.64),(4.63±1.87),(4.03±2.17),(3.32±1.74),(5.30±1.97)points in group A,the difference was statistically significant(F=3.82-7.94,P＜0.05).Conclusions Home diabetes care platform based on internet,combined with family fixed partner education,are more conductive to patient blood sugar control,and enhance self-management ability and account ability.

Objective To translate the Self-Stigma Scale (SSS )into chinese version and check its reliability and validity. Methods The Chinese version of Self-Stigma Scale was translated by Brislin method, the reliability and validity was tested in 750 diabetes patients. Results The Chinese version of Self-Stigma Scale includs 28 items three dimensions. Total scale Cronbach′s α :0.949, all subscales>0.8, Retest reliability coefficient:0.873.The scale average validity index (S-CVI/Ave) was 0.944. Confirmatory factor analysis results show that the GFI: AGFI: 0.833, 0.805, RMSEA: 0.065 the fitting indicators show that the fitting is good. Conclusions The revised Chinese SSS has good reliability and validity. It can be a suitable tool to evaluate stigma level of diabetes patient.

Background:Dissolution therapy is used as an alternative to surgery in patients with gallbladder stone and contraindication for surgery.Finding medicines with optimal litholysis effect and low cytotoxicity is of great importance in clinical practice.Aims:To study the litholysis effect and biological safety of different ethers for gallbladder stones.Methods:Methyl tertiary-butyl ether (MTBE),ethyl tertiary-butyl ether (ETBE),ethylene glycol mono-tert-butyl ether (ETB),and di (ethylene glycol) tert-butyl ether (DETB),arranged by the order of relative molecular weight,were used to dissolve the gallbladder stones in vitro.Normal human liver cell line LO2 was treated with the above mentioned four ethers,and the cell viability was assessed by CCK-8 assay for analyzing the proliferative toxicity.Thirty Sprague-Dawley rats were administered intragastrically with the four ethers for 2 weeks,and then the serum biochemical indices and histopathology of liver,lung,kidney,stomach,duodenum and jejunum were examined.Results:The litholysis effect of MTBE was the most prominent (P ＜ 0.05);that of ETB and DETB were weaker than ETBE (P ＜ 0.05) but still had a substantial effect.CCK-8 assay showed that ETBE was the most toxic ether (P ＜ 0.05),and the proliferative toxicity of ETB and DETB was similar (P ＞ 0.05).Except for the slightly increased serum AST in MTBE,ETBE and ETB groups (P ＜ 0.05),the four ethers had no significant impact on serum biochemical indices and histopathology of multiple organs of the model rats.Conclusions:The increase of relative molecular weight of ether may reduce its litholysis effect,but meanwhile its biological safety is increased.

Objective: To investigate the possibility and diagnostic efficiency of ¹⁸F-NaF PET/CT bone scan after oral administration (PO) by comparing with that of intravenous injection (IV). Methods: Fifty patients (19 males, 31 females; average age: (52.8±11.7) years) with cancer who underwent PET/CT scans after oral and intravenous administration of ¹⁸F-NaF respectively with an interval of 2-7 d from June 2015 to September 2016 were prospectively enrolled. Single-phase ¹⁸F-NaF PET/CT was performed 60 min after IV, and dual-phase ¹⁸F-NaF PET/CT was performed 60 and 120 min after PO. All PET/CT images were reviewed, lesions were counted, and maximum standardized uptake value (SUV_max) and target/non-target (T/NT) ratios were calculated and compared. Paired t test was used. Results: Forty-one patients (15 males, 26 females; average age: (53.5±10.4) years) who finished all PET/CT scans were enrolled. The images at 120 min after PO was visually similar to the images at 60 min after IV. Three modalities detected the same cases and lesions (35 positive cases: 25 malignant, 8 benign, 2 cases with indefinite diagnosis; 302 lesions: 172 malignant, 108 benign, 22 ambiguous lesions). The SUV_{max-PO60 min} and SUV_{max-PO120 min} were lower than the SUV_{max-IV60 min} in the same lesion (18.22±12.64, 26.60±19.49 vs 28.07±16.34; t values: -12.36 and -3.59, both P<0.05). A total of 194 lesions were included for T/NT ratio analysis. T/NT_{IV60 min}, T/NT_{PO60 min} and T/NT_{PO120 min} were 2.76±1.30, 2.87±1.50, 2.98±1.42, respectively, and T/NT_{PO120 min} was higher than T/NT_{IV60 min} (t=3.18, P<0.05). Conclusion ¹⁸F-NaF PET/CT images after PO, especially at 120 min post-PO, has similar diagnostic power of lesion-detection and SUV_max-measurement with IV.

Objective To study the effects of letrozole and clomiphene on the treatment of ovulation induction and the changes of reproductive hormone in the patients with polycystic ovary syndrome.Methods In Zhenhai District People's Hospital from January 2015 to December 2016, 100 cases with polycystic ovary syndrome as the research object in this study, were randomly divided into the control group and the experimental group, 50 cases in each group.The control group were treated with clomiphene, the experimental group were given letrozole combined clomiphene.The levels of reproductive hormones, ovulation rate and pregnancy rate were compared in the two groups.Results The ovulation rate was 90%, pregnancy rate was 46%, the numbers of follicles was(1.2±0.4).In the control group, the ovulation rate was 72%, the pregnancy rate was 22%, and the number of follicles was(2.2±0.5).The results showed that the ovulation rate and pregnancy rate in the experimental group were significantly higher than those in the control group, with statistical significance(P<0.05).The number of follicles in the control group was significantly higher than that in the control group, with statistical significance(P<0.05).There was no significant difference between the experimental group and the control group in the time of follicular maturation, which was not statistically significant.The levels of reproductive hormones in the experimental group were significantly lower than those in the control group, with statistical significance(P<0.05).Conclusion Letrozole combined clomiphene can improve the ovulation rate and pregnancy rate, reduce the reproductive hormone level, which has clinical application value.

Objective To investigate the effect of Daying-35 combined with metformin on ovulation induction after polycystic ovary syndrome, and to summarize the clinical experience.Methods Forty patients with polycystic ovary syndrome were treated with Daying-35 and treated as control group.Forty-five patients were treated with meta-35 combined with metformin and treated as observation group.After treatment, Level and ovulation induction, selected patients were our hospital in January 2016 to December 2016 between the treatment.Results There were no significant differences in T, FSH, LH, E2 and other hormone levels between the two groups before treatment.After treatment, the improvement of the treatment group was significantly more obvious;compared with the two groups of ovarian hyperstimulation syndrome and The rate of pregnancy was 47.5% in the observation group and 15.0% in the control group.The difference between the two groups was significant(P<0.05).Conclusion The effect of Daying-35 combined with metformin on ovulation induction after polycystic ovary syndrome was found to improve the progesterone level and improve the clinical status of the patients.Making the ovulation situation improved, improve the pregnancy rate, it is worth clinical reference.

Objective To explore the application and effect of case-based learning(CBL)in vas-cular surgery clinical teaching. Methods Totally 37 resident doctors were randomly divided into 2 groups respectively: CBL teaching group (n=21)and traditional teaching group (n=16). CBL teaching was con-ducted through the following procedures:selecting typical cases-establishing and applying typical case library-autonomous learning-holding regular seminars. Traditional teaching was conducted through the following procedures: basic theory studying-participating in clinical practice-participating in case discus-sion. Evaluation was conducted based on test socre (written test and clinical operational skill test)and res-idents' feedback of teaching effect. Data were statistically described and independent sample t test was performed. Results Theoretical exam score and clinical skill test score were high in CBL group than in traditional group ((thoretical score:(85.53 ±1.75) vs. (79.94 ±2.29);clinical skill test score:(85.10±1.64)vs.(80.31±1.82)). CBL teaching group had advantages in improving learning efficiency, cultivat-ing clinical thinking,promoting mastery and application of knowledge,broadening knowledge, promoting communication and expression ability and improving study enthusiasm ,et al . Conclusion CBL teaching can effectively improve the teaching quality and obtain higher evaluation. Typical case li-brary should be constantly improved and education of vascular surgical basic theory should be strength-ened to promote CBL.

Objective To analyze, treatment of ipsilateral Pilon and calcaneal fractures (IPCF). Methods Injury causes included fallings in seven patients, traffic injury in two and impact injury in one, with ISS score of 5-22 points. There were seven patients with open injuries and 3 with close ones. Nine pa-tients had Ⅱ-Ⅲ degree of Pilon fractures according to the AO/OTA classification and six comminuted com-pression calcaneal fractures according to the Essex-Loprest classification. The Pilon fractures were fixed by cannulated nail, screw or Kirschner wire through open reduction in six patients including four fixed with ex-ternal fixators (the calcaneal fractures were fixed with percutaneous eannulated nail or Kirschner wire by poking reduction). The calcaneal fractures were fixed with only Kirschner wire through poking reduction in two patients and double fractures treated conservatively in two. Results The reduction of Pilon and cal-caneal fractures treated by operation were markedly improved in six patients, while there occurred angular deformity and joint surface introeession in the patients treated conservatively. The wound healing of soft tis-sue was sound in seven patients but unsatisfactory in the other three. The ankle-hind foot score was 82-94 points in four patients who were followed up. Conclusions IPCF is caused by the high energy force and characterized by severe injures of the soft tissues and the bone tissues at the distal tibia and calcaneua, for which limited internal fixation plus external fixation are suitable treatment choice.