In-silico annotation of the chemical composition of Tibetan tea and its mechanism on antioxidant and lipid-lowering in mice.

BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice. MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH2O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected. Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS: This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

Effects of Pu-erh and Dian Hong tea polyphenols on the gut-liver axis in mice.

Tea polyphenols (TP) are the most biologically active components in tea, with antioxidant, antiobesity, and antitumor properties, as well as the ability to modulate the composition and function of intestinal microbiota. This experimental study evaluated the chemical constituents of polyphenols in Pu-erh (PTP) and Dian Hong tea (DHTP). It also investigated the co-regulatory effects of PTP and DHTP on intestinal flora and liver tissues in mice using 16 S rRNA gene and transcriptome sequencing. The results revealed that DHT had higher concentrations of EGC (epigallocatechin), C (catechin), EC (epicatechin), and EGCG (epigallocatechin gallate). In contrast, PT had higher concentrations of GA (gallic acid), ECG (epicatechin-3-gallate), TF (theaflavin), and TB (theabrownin). PTP and DHTP consumption significantly reduced the rates of weight gain in mice. Microbial community diversity was significantly higher in PTP and DHTP-treated mice than in the control group. Notably, beneficial microbes such as Lactobacillus increased significantly in PTP-treated mice, whereas Lachnospiraceae increased significantly in DHTP-treated mice. Both PTP and DHTP improved the activity of the antioxidant enzymes (SOD) and total antioxidant capacity (T-AOC) in the liver. The transcriptome analysis revealed that the beneficial effects of PTP and DHTP were due to changes in various metabolic pathways, the majority of which were related to antioxidant and lipid metabolism. This study discovered that PTP and DHTP had beneficial effects in mice via the gut-liver axis.

Metabolomic Analysis of Fermented Tibetan Tea Using Bacillus circulans and Their Biological Activity on Mice via the Intestine-Hepatic Axis.

The use of Bacillus circulans as the sole starter provides better process control compared to natural fermentation. However, the chemical composition of fermented Tibetan tea by B. circulans and its regulatory effects on the intestine-liver axis has not been reported. For this purpose, a high-resolution liquid chromatography tandem mass spectrometry metabolomics approach was performed. The effects of fermented Tibetan tea on the intestine-liver axis of mice were also evaluated. Untargeted metabolomics analysis showed that the contents of catechin derivatives, flavonoids, phenolic acids, and terpenoids increased by 0.3, 2.38, 2.65, and 3.36%, respectively, compared with those before fermentation. Furthermore, 16S ribosomal RNA sequence analysis revealed that the relative abundance of Lactobacillus spp. in the intestine increased after consumption of fermented tea. Additionally, based on histological and quantitative PCR analyses, fermented Tibetan tea also improved intestinal development and intestinal barrier function in mouse, while increasing the antioxidant capacity of mouse liver. Thus, fermented Tibetan tea could provide beneficial health effects through the intestine-liver axis. These findings have facilitated the study of the chemical composition of Tibetan tea and provided theoretical support for its use as a natural beverage with intestinal probiotic functions.

Multiple Organ Dysfunction Syndrome Caused by Sepsis: Risk Factor Analysis.

PURPOSE: To analyze the risk factors of multiple organ dysfunction syndromes (MODS) caused by sepsis. PATIENTS AND METHODS: A total of 180 patients with sepsis admitted to The First Affiliated Hospital of Harbin Medical University (No. 23, Post Street, Nangang District, Harbin 150001, Heilongjiang province, China) from July 2018 to June 2019 were selected and divided into a non-MODS group and a MODS group, with 90 cases in each group. Clinical data of the patients were retrospectively analyzed, and univariable and multivariable analyses were performed. RESULTS: The univariable analysis showed that there were no significant differences in terms of age, body temperature, heart rate, respiration, mean arterial pressure, RBC specific volume, blood sodium, serum kalium, and infection site (P > 0.05). Whereas significant differences were found between the groups in terms of gender, arterial blood pH, WBC count, Apache II score, blood glucose, creatinine, chronic medical history, surgery, and ventilator usage (P < 0.05). The growth of bacterial culture, the increase of creatinine level, chronic diseases and Apache II score were discovered to have significant effects on the occurrence of MODS through the multivariable logistic regression analysis. CONCLUSION: Bacterial culture, serum creatinine level, history of chronic disease and Apache II score may be risk factors of MODS in sepsis patients.

The Effect of Auricular Plaster Therapy on Insomnia in Patients with Rheumatoid Arthritis.

Objective: This research was conducted to observe the effects of auricular plaster therapy on insomnia in patients with rheumatoid arthritis (RA). Materials and Methods: This study involved 76 patients with insomnia caused by RA, who were admitted to Foshan Chancheng Central Hospital-in Foshan, Guangdong, China-from August 2017 to August 2018. The patients were randomized, with 38 to the intervention group and another 38 to the control group. The intervention group was treated with auricular plaster therapy with beans, while the control group was treated with estazolam orally before going to bed. Curative effects were compared between the 2 groups. The Pittsburgh Sleep Quality Index scale (PSQI) and the Athens Insomnia Scale (AIS) were used to assess and evaluate the sleep quality of patients. Result: Scores of the PSQI dimensions all fell in both groups after treatment. Scores for sleep quality, sleep latency, sleep efficiency, sleep disorder, and daytime dysfunction in the observation group were lower than those in the control group (P < 0.05). AIS scores in all dimensions decreased in both groups, and, after treatment, scores and total points of sleep latency, night-time revival, overall sleep quality, and the dimensionality of daytime body function in the intervention group were lower than those in the control group (P < 0.05). Conclusions: Auricular plaster therapy has a marked effect on insomnia in patients with RA, and the therapy is easy and simple to apply.

[Study on clinical value of three localization methods in laparoscopic colorectal tumor surgery].

OBJECTIVE: To evaluate the clinical effectiveness of three localization methods, including methylene blue, metal clips and intraoperative colonoscopy in laparoscopic colorectal surgery. METHODS: A retrospective analysis was performed to review the clinical data of 64 patients who underwent the laparoscopic colorectal operations in Cancer Hospital of Fudan University from December 2009 to June 2012. Three methods of tumor localization were used perioperatively, including 23 cases of methylene blue, 20 of metal clips and 21 of colonoscopy. RESULTS: Operations were successfully performed in this cohort and there were no deaths or complications. In methylene blue group, intraoperative colonoscopy was performed in two cases because of the inability to visualize blue dye on the serosal surface of the intestinal wall, another 2 cases were converted to open operation because of methylene blue diffusion and inability to identify resection margin. Intraoperative colonoscopic localization was required for 3 cases of sigmoid colon or upper rectal tumor because of inaccurate tumor localization by metal clips. Poor operative exposure due to obvious bowel distension prompted the conversion to open surgery in 2 cases of colonoscopy localization group, and the accurate position of the lesion was not found in another 2 cases due to long pedunculated adenoma. CONCLUSIONS: Colorectal tumor can be localized effectively by endoscopic methylene blue tattooing at a maximum of 2 tumors before operation and the method of 4-point positioning can significantly improve the accuracy of colorectal tumor localization. Tumor localization preoperatively on the day of surgery by metal clip is accurate for the right or left colon cancer. Intraoperative colonoscopy can localize tumor accurately and rapidly for rectosigmoid or descending tumor, and the incidence of bowel distension can be significantly reduced. Localization method should be considered according to the tumor location and surgical procedure.

[Clinical analysis of 168 cases of multiple primary colorectal carcinoma].

OBJECTIVE: To study the incidence rate of multiple primary colorectal carcinomas (MPCC) in colorectal carcinoma and to evaluate its clinical and pathological characteristics. METHODS: One hundred and sixty-eight (4.6%) patients from 3663 cases with colorectal carcinoma were diagnosed with MPCC from January 1985 to December 2003. The clinical data of the patients were collected retrospectively to investigate the diagnosis and treatment of MPCC. RESULTS: Of the 168 patients, 81 were diagnosed as synchronous colorectal carcinoma (SC), 72 with metachronous colorectal carcinoma (MC), 15 with both SC and MC. The median age at time of diagnosis of colorectal carcinoma was 58 years old (range from 20 to 82 years old). Three hundred and ninety-three cancer lesions were detected in these 168 cases (mean, 2.3 lesions/case). The rectum and sigmoid colon were the most involved sites (61.6%). Eighteen cases (10.7%) were verified with hereditary non-polyposis colorectal cancer (HNPCC) while another 9 cases were highly suspected. Fourteen patients (8.3%) were found with other malignancies out of large intestine, 41 patients (24.4%) with colorectal adenomas, 72 (42.9%) with adenoma carcinogenesis. Among the 96 SC patients, 91 were given preoperative colonoscopy and 65 (71.4%) got the diagnosis. All the MC patients were diagnosed by postoperative colonoscopy. The overall 5-year survival rate of the 168 patients was 69.8%. CONCLUSIONS: MPCC should be paid more attention in colorectal cancer management. Colonoscopic surveillance is much more important in diagnosis and follow-up of MPCC for reducing the misdiagnosis of SC and detecting more MC in time. Prompt treatment of adenoma can reduce the occurrence of MPCC, and active and standard surgical treatment should be done for MPCC.

[Study on the germline mutation of MSH6 gene in Chinese hereditary nonpolyposis colorectal cancer pedigrees using PCR based sequencing].

OBJECTIVE: To detect the germline mutation of mismatch repair gene (MSH6) in hereditary nonpolyposis colorectal cancer (HNPCC) kindreds fulfilling different clinical criteria. METHODS: The germline mutations of MSH6 gene were detected by PCR based DNA sequencing in 39 unrelated HNPCC probands fulfilling different clinical criteria in which MSH2 and MLH1 mutations were excluded. The exons with missense mutations were analyzed using PCR sequencing in the germline genomic DNA of 137 healthy persons. The expression of MSH6 protein was detected by Envision immunohistochemistry staining in the tumor tissues of the mutational probands. RESULTS: Six germline mutations of MSH6 gene were detected in 39 probands of Chinese HNPCC kindreds, and the mutations distributed in the exon 4, 6, 9 and 10. Four out of six mutations were missense mutation, one was nonsense mutation and the remaining one was insertion mutation in splice site. The results of sequecing for the exons with above four missense mutations in 137 healthy persons' genomic DNA showed that 5 of 137 persons had the missense mutation of c.3488 A to T at codon 1163 of the 6th exon. The mutational rate was approximately 3.65% (5/137), so the mutation could be a single nucleotide polymorphism (SNP). The remaining missense mutations were not found in any germline genomic DNA of 137 healthy persons. Positive expression of MSH6 protein had been identified in the tumor of the SNP proband while the tumors had negative MSH6 protein expression in the rest probands of germline mutation MSH6 gene. The types of mutations and their potential significance were determined by comparing the following databases: http://www.ncbi.nlm.nih.gov/, http://www.ensembl.org/homo-sapies, and http://www.insight-group.org. Five out of the six mutations had not been reported previously and they were new pathological mutations, the rest one was a new SNP. CONCLUSION: Germline mutations of MSH6 gene may play an important role in Chinese HNPCC kindreds fulfilling different clinical criteria. It is necessary to analyze the germline mutations of MSH6 gene using sequencing to identify HNPCC families in the probands in which MSH2 and MLH1 mutation were excluded.

Effect of short-chain fatty acids on the proliferation and differentiation of the human colonic adenocarcinoma cell line Caco-2.

OBJECTIVE: Short-chain fatty acids (SCFA) in the colon may maintain colonocyte differentiation and oppose carcinogenesis. The purpose of this study was to investigate the effects of three SCFA, butyrate, propionate and acetate, on the differentiation, proliferation, and matrix interactions of the Caco-2 human colonic adenocarcinoma cell line. METHODS: Differentiation was assessed by brush border enzyme expression and the doubling time (proliferation) was calculated directly from serial cell counts and by the logarithmic transformation method. Cell motility (migration) was quantitated by the expansion of a confluent Caco-2 monolayer (after release from a constraining fence) across bacteriologic plastic dishes precoated with saturating concentrations of type I collagen. Results were expressed as mean +/- SE and were analyzed by ANOVA and Bonferroni's modified t-test. RESULTS: All three SCFA studied altered the Caco-2 phenotype. Treatment with 10 mmol SCFA significantly prolonged the cell doubling time, promoted brush border enzyme expression (cathepsin C), and inhibited the motility of the Caco-2 cells. CONCLUSIONS: Butyrate, propionate and acetate inhibited the proliferation and motility of a well-differentiated human colonic cancer cell line while promoting the expression of the differentiation marker, cathepsin C. Thus the SCFA produced by bacterial fermentation of dietary fiber may exert a protective effect against the development of colon cancer.

[Discussion for clinical treatment in early stage of colorectal carcinoma with the invasion limited in the layer of muscularis mucosae].

OBJECTIVE: To emphasis the importance of recognizing and treating the early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae, and discuss several questions in using the WHO's new diagnostic standards of the prestage of tumors of digestive system. METHODS: A retrospective review was made of 30 patients with invasive carcinoma limited in the layer of muscularis mucosae. All of the patients were treated between July 1986 and July 1999 in Shanghai Cancer Hospital. RESULTS: Among the 30 cases, there were 19 men and 11 women from 20 to 80 years dd (mean, 57.0). The tumor diameter was ranged from 0.8 cm to 8.0 cm (mean, 2.9 cm). 12 patients were given polypectomy while 18 patients were given radical resection. Among the patients given polypectomy, one got recurrence 3 years later, one had the metastasis of lungs 50 months later and had been dead already. Among the patients given radical resection, one had metastasis of lymph nodes peripheral to the tumor, one also had the metastasis of lungs 68 months later but is still alive now, and one had metastasis of lymph nodes in the right upper part of neck 15 months later and then was lost to follow-up. CONCLUSIONS: The patients with early colorectal carcinoma with the invasion limited in the layer of muscularis mucosae could have metastasis of lymph nodes, get local recurrence after polypectomy and even have blood metastasis. Therefore great attention must be paid. Clinicians should be careful to chosen polypectomy. Meanwhile, in order to provide more information of the correct treatment, it will be better if the pathologists could give the original diagnosis at the same time when they use the WHO's new diagnostic standards of the prestage of tumors of digestive system in diagnosing the early invasive carcinoma limited in the layer of muscularis mucosae, for the concept of high-grade intraepithelial neoplasia will be used instead of it, and whether or not it is a really benign tumor, much more clinical research must be done later.

[Realization of clinical diagnoses and treatment for dermatofibrosarcoma protuberans].

OBJECTIVE: To emphasize the importance of correct and standardized surgical treatment on dermatofibrosarcoma protuberans (DFSP), and discuss the suitable synthesized therapy on it. METHODS: 163 cases of DFSP, which were treated between January 1985 and September 2002,were submitted to a retrospective study. RESULTS: Among the 163 cases, 150 (92.0%) were treated with local excision as benign tumors before accepted to Cancer Hospital, Fudan University. 69 cases (46.0%) were approved by pathological examination to have tumor remnants after they were treated with wide excision, and 49 (71.0%) of them couldn't be found to have any tumor remnants by physical examination or B-ultrasonic examination before that operation. It was easy for the tumor to recur after excision, especially the local excision. The recurrent rate after it was 45.1%, which was much higher than the one after wide excision (5.6%). Among the 142 cases which wide excision were performed, 99 ones had excision margins >/= 3 cm and 5 of them (5.1%) developed local recurrence while 36 ones had excision margins 1 approximately 2 cm and 3 of them (8.3%) developed local failure. 46 cases (32.4%) were given skin graft, 11 cases were given flap, and 1 case had dacron mending in skin defection area. The main complications after these operations were necrosis of the skin flap (20 cases) and infection of the wound (6 cases). They could all be cured in 2 months. 17 cases were given complimentary radiotherapy with the dose range from 3275 cGy to 7000 cGy because of their recurrences for times or positive resection margins after wide excision. Only one case had wet molting after radiotherapy and 2 developed local recurrence. Among all the 163 cases, only 2 (1.2%) were dead, and 1 of them was died of metastasis of lung and liver. 2 cases got lymph node metastasis, then were given surgical treatment and still alive now. 13 cases (8.0%) were DFSP-FS with their malignancies increased. 11 of them were the recurrent ones after local excision or wide excision. CONCLUSIONS: In order to avoid misdiagnosis, it is necessary for the clinician to know much about DFSP. Once the tumor was diagnosed of DFSP after local excision, it is necessary to take wide excision. Because DFSP is a malignancy of a high recurrent rate after local excision, standardized wide excision is the key in reducing local failure. Adjuvant radiotherapy is an effective treatment for the patients with positive resection margin or the patients don't suit for surgical treatment. The DFSP-FS need to use more energetic treatment in curing it.

[Mutation analysis of hMSH2 and hMLH1 genes in Chinese hereditary nonpolyposis colorectal cancer families].

OBJECTIVES: To determine the germ-line mutations of hMSH2 and hMLH1 genes in Chinese hereditary nonpolyposis colorectal cancer (HNPCC) families' probands or in patients fulfilling different clinical criteria or guidelines; to clarify the nature and distribution of the mutations; to evaluate the sensitivity of different clinical criteria in mutation prediction. METHODS: The entire coding regions (35 exons including exon-intron boundaries) of hMSH2 and hMLH1 genes were directly sequenced in 24 Amsterdam criteria (AC) probands, 15 Japanese criteria (JC) probands (except AC kindreds) and 19 Bethesda guidelines (BG) patients (except two former groups). All available affected and unaffected members from families of those with mutations were screened for mutation. RESULTS: In 16 unrelated families selected by the different clinical criteria, 17 germ-line mutations were found with 11 (64.7%) of hMLH1 and 6 (35.3%) of hMSH2. Two mutations were identified in one of the families. Among the 17 germ-line mutations, 12 had not been reported previously. A diversified mutation spectrum was found, but 6 hMLH1 mutations were found to be concentrated in the region encompassing exon 14, 15 and 16. There was a wide spectrum of mutation type including frame shift, nonsense, splice site mutation, in frame insertion or deletion and missense mutations. The mutation detection rate of hMSH2 and hMLH1 in the AC group was significantly higher than that in the JC group (12/24 vs. 3/15). On the other hand, a low mutation rate (1/19) was detected in 19 BG patients. The mutation cosegregated with disease. Besides, three different genotypes in tumors from probands of mutation-positive families were found. CONCLUSIONS: hMSH2 and hMLH1 mutations in Chinese HNPCC families show a wide spectrum. It seems that hMLH1 gene is involved more frequently than hMSH2 gene in Chinese HNPCC families. Different clinical criteria predict mutations with different sensitivities. The Amsterdam Criteria are most sensitive, while Japanese Criteria are highly practical and the Bethesda Guidelines are also practical to some extent. Gene mutations cosegregate with the disease phenotype. Carriers with no symptom in HNPCC families are most vulnerable groups, follow-ups are required for this group to get early diagnosis and to prevent the development of CRCs.

Clinical characteristics and diagnosis of patients with hereditary nonpolyposis colorectal cancer.

AIM: To study the clinical characteristics of hereditary nonpolyposis colorectal cancer (HNPCC) in the Chinese population and discuss the identification and management of the patients with HNPCC. METHODS: A series of 140 patients with colorectal cancers (CRC) and HNPCC associated tumors from 30 families fulfilling the Amsterdam criteria were analyzed. RESULTS: A total of 118 patients had CRC. Average age at diagnosis of the first CRC was 45.7 years, 56.8 % and 23.4 % of the first CRC were located proximal to the splenic flexure and in the rectum respectively. Twenty-three (19.5 %) had synchronous and metachronous CRC. Twenty-seven patients were found to have extracolonic tumors. Gastric carcinoma was the most common tumor type in our series (44.4 %). CONCLUSION: The frequency of HNPCC was 2.6 % in our series of patients. The main features are an excess of early onset with a propensity to involve the proximal colon, and high frequency of multiple foci. Management and surveillance for these patients should be different from sporadic CRC. Contrary to American and European reports, gastric cancer seems more frequent than endometrial cancer in Chinese. It is necessary to formulate a new HNPCC criterion for Chinese patients.