[Characteristics and clinical analysis of MLH1 c.463dupC gene mutation in a Lynch syndrome family].

In this study, a case of Lynch syndrome (LS) family line with a novel mutation site in the MLH1 c.463dupC gene was reported and the clinical and pathogenic genetic features of this family were analyzed. A 40-year-old female patient with colon cancer diagnosed at the First Affiliated Hospital of Kunming Medical University on October 2, 2020 was retrospectively included. The clinical data of the family were collected and the family lineage was drawn. The family tumor history met the Amsterdam Criteria Ⅱ and the diagnostic criteria of LS in Chinese, which was a typical LS family lineage. A germline code-shift missense mutation c.463dupC in the MLH1 gene located in exon 6, a possible pathogenic variant, was detected by second-generation sequencing (NGS) in the patient. Subsequently, Sanger sequencing was performed on a total of 20 direct lineage members of the family of the MLH1 gene, 7 cases were found to harbor the mutation and included in the LS high-risk control. Follow-up to October 2023 showed that the patient had endometrial and cervical polyps, one case had colorectal cancer, and two cases had intestinal polyps, all were treated with early intervention and therapy; two cases did not show any clinical symptoms. This study is the first to report a new mutation site for the potentially pathogenic MLH1 c.463dupC, providing a rationale for the pathogenicity of the mutation and standardized health management for familial carriers.

[Clinical effect of free thoracodorsal artery perforator flap in reconstructing large scar on the facial subunit].

Objective: To explore the clinical effect of free transplantation of thoracodorsal artery perforator flap in reconstructing large scar on the facial subunit. Methods: From April 2014 to March 2018, 7 patients with large facial scar were admitted to Ningbo NO.6 Hospital, including 3 males and 4 females, aged from 31 to 49 years, 4 with frontal involvement and 3 with chin and neck. Color Doppler ultrasound was used for the positioning of the thoracodorsal artery perforating vessel, and scar resection was performed according to the principle of facial subunit repair. The wound area was 8 cm×6 cm-21 cm×8 cm, and the wound was repaired with the free thoracodorsal artery perforator flap in the area of 9 cm×7 cm-22 cm×9 cm. The donor site was closed directly by suturing. The consistency of the location of the perforating vessel explored during the operation with its preoperative positioning and the flap survival were recorded. The color, texture, and appearance of the flap and the healing condition, scar formation, and function of the donor area were observed during follow-up. Results: The locations of the perforating vessels of 7 patients explored during the operation were consistent with those positioned by color Doppler ultrasound before the operation. All the flaps of the 7 cases survived successfully after operation. Postoperative follow-up of 12-18 months showed that the flap color was similar to the surrounding skin of the recipient area, with soft texture and no obvious contracture. Slight bloated appearance was observed in the flaps of 4 cases. All the 7 patients had postoperative healing of the flap donor site without obvious scar hyperplasia or influence on shoulder joint function. Conclusions: The anatomy of the perforating vessel of the thoracodorsal artery perforator flap is relatively constant and the flap can be cut in large area with soft texture, good appearance, and concealed donor area, which is a good choice for reconstructing large scar on the facial subunit.

Long non-coding RNA DILC as a potentially useful biomarker for the diagnosis and prognosis of colorectal cancer.

OBJECTIVE: LncRNA downregulated in liver cancer stem cells (lnc-DILC) has been implicated as a tumor suppressor in colorectal cancer (CRC). However, the clinical significance of lnc-DILC in CRC patients has not been investigated. In this study, we aimed to explore the diagnostic and prognostic value of lnc-DILC in CRC patients. PATIENTS AND METHODS: The expression of lnc-DILC was measured in 174 paired CRC tissues and adjacent normal tissues using Real Time-Polymerase Chain Reaction (RT-PCR). The correlation of lnc-DILC expression with clinicopathological factors was statistically analyzed by the Chi-square test. Besides, overall survival analysis was carried out with the Kaplan-Meier curve with the log-rank test. Univariate and multivariate analyses were performed to explore the prognostic significance of lnc-DILC expression. RESULTS: We found that lnc-DILC expression was downregulated in CRC tissues compared to their adjacent normal tissues (p<0.01). ROC analyses showed that lnc-DILC levels were reliable in distinguishing patients with CRC from normal colorectal tissues. Then, down-regulation of lnc-DILC was positively associated with aggressive clinical characteristics, including depth of invasion (p=0.018) and advanced TNM stage (p=0.009). Moreover, the Kaplan-Meier analysis demonstrated that overexpression of lnc-DILC was associated with poorer overall survival (p=0.0205) and disease-free survival (p<0.001). Finally, multivariate analyses confirmed that expression of lnc-DILC was an independent prognostic factor in CRC. CONCLUSIONS: Our results firstly suggested that lnc-DILC could be a favorable indicator of prognosis in CRC.

[Clinical and genetic analysis of an infant with isolated 17, 20-lyase deficiency].

OBJECTIVE: To explore the clinical and genetic characteristics of an infant with isolated 17, 20-lyase deficiency. METHOD: The clinical, biochemical and genetic characteristics were analyzed in an 8-month-old infant with 46, XY gonadal dysgenesis who presented predominantly the female external genitalia. RESULT: The infant was referred because of"masses in bilateral inguinal region and 46, XY gonadal dysgenesis". He was normotensive. Laboratory tests revealed elevated levels of progesterone and 17-hydroxyprogesterone. The detailed parameters are as follows: progesterone 29.35(reference range 0.09-1.0)nmol/L, 17-hydroxyprogesterone 10.9(reference range 0.6-2.6)nmol/L, testosterone 0.7(reference range 0.1-3.1)nmol/L, dehydroepiandrosterone sulfate <0.15(reference range 0.80-5.6)mg/L, androstenedione <0.3 (reference range 0.6-3.1) µg/L, luteinizing hormone 6.6(reference range 0.6-1.7)U/L, follicle stimulating hormone 1.8 (reference range 0.5-3.7)U/L, estradiol 37.66(reference range 73.4-146.8)pmol/L. The patient had normal levels of serum sodium, potassium, corticosteroid and plasma adrenocorticotropic hormone. Genomic DNA was extracted from the leukocytes of peripheral blood of the patient and subjected to next generation sequencing (NGS) for testing more than 200 sexual development related genes. Sanger sequencing was used to confirm the results of NGS. Genetic analysis revealed that the patient harbored compound heterozygous mutations of c. 1226C>G (p.Pro409Arg, P409R) and c. 707T>G (p.Val236Gly, V236G) in CYP17A1 gene derived from paternal and maternal allele. V236G was a novel mutation predicted to be pathogenic. The infant was diagnosed as isolated 17, 20-lyase deficiency combined with clinical and molecular characteristics of CYP17A1 gene. CONCLUSION: We have identified the compound heterozygous mutations of P409R and V236G in the CYP17A1 gene in one infant with isolated 17, 20-lyase deficiency. He presented with 46, XY gonadal dysgenesis, normal blood pressure and elevated concentration of progesterone and 17-hydroxyprogesterone.

Construction and selection of the natural immune Fab antibody phage display library from patients with colorectal cancer.

AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected. RESULTS: The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6). The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer. CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.

Curcumin inhibits cell proliferation by interfering with the cell cycle and inducing apoptosis in colon carcinoma cells.

It has been reported that curcumin (diferuloylmethane) could inhibit growth of several types of malignant cells both in vitro and in vivo. However, the mechanism of its action is unknown. In this study, we investigated the inhibitory effects of curcumin on human colon carcinoma cell (Lovo) growth and its mechanism of action in vitro by means of growth assay, colony formation assay, MTT assay, cell cycle and apoptosis analysis. Curcumin inhibited cell growth in a dose-dependent manner. The ability of Lovo cells treated with curcumin to form colonies was depressed. MTT test showed that curcumin was cytotoxic to cells. Lovo cells treated with curcumin were largely accumulated in S, G2/M phase which prevented cells from entering the next cell cycle. Apoptosis induced by curcumin was confirmed by characteristic ladders and cellular morphological changes. Curcumin can inhibit Lovo cells growth and the cellular mechanism responsible for the action is to arrest the cell cycle in S, G2/M phase and to induce apoptotic cell death.

Comparison of preoperative TN staging of gastric carcinoma by endoscopic ultrasonography with CT examination.

AIM: To assess the accuracy and limitations of endoscopic ultrasonography (EUS) in the preoperative staging of gastric carcinoma in comparison with computed tomography (CT). METHODS: According to the new (1987) TN staging, 62 patients with gastric carcinomas were examined preoperatively by EUS and the results compared with those of postoperative pathological TN staging. CT of abdomen was performed before surgery for 32 of the patients. RESULTS: The overall accuracy of T staging was 83.9% for EUS and 28.1% for CT. For the detection of regional lymph node metastases, EUS accuracy was 79.0%, sensitivity 80.0% and specificity 87.5%, versus 50.0% accuracy for CT. The coincidence of perigastric infiltration was 90.0% for EUS and 41.2% for CT. The most frequent causes of misdiagnosis by EUS were microscopic tumor invasion and peritumorous inflammatory or fibrous changes. CONCLUSION: EUS is a reliable method for the clinical evaluation of locoregional extension of gastric cancer and more accurate than CT in the preoperative staging of gastric carcinoma.

[A study on the complementary scheme of mass screening for colorectal cancer in an asymptomatic population].

Both SPA immunological fecal occult blood test (SPA FOBT test) and detection of T-antigen in rectal mucus (Shams' test) were used as screening tests in asymptomatic mass screening to evaluate the complementary effect of both tests for detection of colorectal cancers. SPA FOBT test showed a positive rate of 11.1% and shams' test 8.9% among 7,740 subjects. In 498 cases with positive screening test, 11 cases of carcinomas and 88 adenomas were found with colonoscopy. Only 9 cancers and 55 adenomas showed positive result in SPA FOBT test and 8 cancers and 51 adenomas in Shams' test. Both tests combined could enhance the detective rate of cancer in asymptomatic mass screening from 81.8% with SPA FOBT test or 72.7% with Shams' test to 90.9%. This complementary effect was more obvious in adenoma detection. It is suggested that there were some shortcomings in sequential FOBT test for cancer detection, the combined use of the two different screening tests for detection of colorectal cancer could decrease the rate of missed detection.

Comparison of Shams' test for rectal mucus to an immunological test for fecal occult blood in large intestinal carcinoma screening. Analysis of a check-up of 6480 asymptomatic subjects.

The sugar moiety detected from rectal mucus by the Galactose oxidase-schiff (Shams' test) is considered a substitutive test for immunological fecal occult blood test (FOBT) in screening colorectal carcinoma. Two strategies of screening were applied in 6480 subjects over 40 years of age, and 130 cm flexible colonoscope used for sigmoidoscopy or pancolonoscopy. Of them, 3820 were taken for immune FOBT (SPA test) and Shams' test. Only those who showed positive tests were chosen for 60 cm flexible sigmoidoscopy, while another 2660 subjects for both sigmoidoscopy and tests at the same time. Additionally, 130 cm flexible pancolonoscopy was carried out in 103 individuals with positive Shams' test for evaluating the false positive rate. Shams' test showed a sensitivity of 85.7% for colorectal cancer, 47.1% for adenomas in preselected patients, while the positive rate of SPA test were 90.5% and 41.2% respectively. In 3820 asymptomatic subjects undergoing sequential screening (aged 45 years and higher), Shams' test showed 9.1% positive, SPA showed 11.2% and 620 (16.2%) subjects were selected for sigmoidoscopy based on their positive galactose oxidase result or positive FOBT result. Two early stage carcinomas and 33 adenomas (0.32% and 4.2% respectively in sigmoidoscopy) were found. Another 2 660 subjects were taken for sigmoidoscopy screening. Four carcinomas and 78 adenomas were found. Of them, only two carcinoma (50%) and 17 (21.8%) or 22 (28.2%) adenomas were positive in Shams' or SPA test. But both tests combined in screening showed a rate of 61.3% in adenomas and 75.0% in cancers. 103 subjects with positive Shams' test were taken for pancolonoscopy. 82.5% showed no lesions.(ABSTRACT TRUNCATED AT 250 WORDS)

Study of molecular markers of resistance to m-AMSA in a human breast cancer cell line. Decrease of topoisomerase II and increase of both topoisomerase I and acidic glutathione S transferase.

Resistance to 0.8 microM 4'-(9-acridinylamino)methanesulphon-m-anisidide (m-AMSA) was induced by stepwise increases of drug concentration in the human tumor cell line CALc18 originating from a breast adenocarcinoma. The resistant cell line CALc18/AMSA exhibited a resistance index of 10 and a cross-resistance to other topoisomerase II inhibitors. A 3-fold decrease in the levels of topoisomerase II decatenating activity was found in CALc18/AMSA cells. By contrast, topoisomerase I activity was increased by about 3-fold in resistant cells. Interestingly this line was hypersensitive to camptothecin, a specific inhibitor of topoisomerase I. Restriction endonuclease patterns of the topoisomerase I and topoisomerase II loci were found to be identical in CALc18/AMSA and CALc18 with no evidence of gene amplification and rearrangements. Alkaline elution of m-AMSA-treated cells showed that DNA single strand breaks and DNA-protein crosslinks were decreased in CALc18/AMSA. The DNA lesions also obtained in m-AMSA-treated nuclei indicated that no drug uptake modification occurred in both cells. Moreover, the in vitro m-AMSA-induced DNA cleavage per unit of decatenating activity and the inhibitory effects of antitumoral drugs on decatenation were not found to be different with topoisomerase II from sensitive or resistant cells. However the specific cleavage induced by m-AMSA/per mg of crude protein from resistant cells was 2 to 3 times decreased. Multidrug resistance gene transcripts were not detected while levels of acidic glutathione S transferase mRNA were found to be 8 to 10-fold greater in resistant than in sensitive cell line with no amplification of the gene. In conclusion, the diminution of topoisomerase II activity and the increase of both topoisomerase I and acidic glutathione S transferase transcripts could contribute to the resistant phenotype of these breast cancer cells.