[Epidemiological investigation of constipation in urban areas of Hangzhou, China].

Objective: To determine the prevalence and risk factors of constipation in Hangzhou urban areas by analyzing data from a recent epidemiological survey. Methods: From August 2022 to June 2023, a cross-sectional survey was conducted on 43 communities on eight streets in suburbs of Hangzhou, including Wangjiang, Sijiqing, Puyan, Changhe, Ziyang, Xiaoshanchengxiang, Zhuantang, and Banshan. Written questionnaires were filled out face to face by community doctors. The prevalence of constipation was investigated using a combination of the Bristol stool typing method and the Rome IV criteria. The risk factors for constipation were analyzed using multivariate logistic regression analysis. Results: The study cohort comprised 10,479 participants, 5551 of whom were male (53.0%) and 4928 female (47.0%). The overall prevalence of constipation was 10.6% (1107/10,479). The prevalence was 11.0% (613/5551) for male and 10.0% (494/4928) for female participants; this difference is not statistically significant (P>0.05).There were no significant differences in the prevalence of constipation between different categories of body mass index (P>0.05). The prevalence of constipation was highest in individuals aged 70 years and above, accounting for 15.4% (264/1717) of all cases. There were significant differences in the prevalence of constipation between different occupations, marital statuses, and educational levels (P<0.001). Patients with hypertension or diabetes mellitus had a higher prevalence of constipation than those without these conditions, those who had been taking long-term medication had a higher prevalence of constipation than those who did not, parents of children with constipation had a higher prevalence of constipation than those with children who did not have constipation, and there were other significant differences in various variables (P<0.001). Multivariate logistic regression analysis of modifiable factors revealed that eating less fruit (OR=1.284, 95%CI: 1.061-1.553, P=0.010), liking spicy and salty food (OR=1.234, 95%CI: 1.039-1.466, P=0.016), sleeping for too long (OR=1.644, 95%CI: 1.260-2.144, P<0.001), irregular sleep patterns (OR=1.370, 95%CI: 1.127-1.665, P=0.002) and minimal exercise (OR=1.388, 95% CI: 1.168-1.649, P<0.001) were all risk factors for constipation; whereas mix diets (OR=0.709,95%CI:0.608-0.826, P<0.001), meat diets (OR=0.604,95%CI: 0.495-0.736, P<0.001), moderate alcohol consumption (OR=0.659, 95% CI: 0.534-0.812, P<0.001), less or no drinking (OR=0.523, 95% CI: 0.428-0.638, P<0.001), and non-smoking (OR=0.819, 95%CI:0.674-0.995, P<0.001) protected against constipation. We found no association between dietary preferences and the risk of constipation (refined grains: OR=1.147, 95%CI:0.944-1.393, P=0.167; no specific preference: OR=0.935, 95%CI:0.783-1.117, P=0.459). The rate of agreement between self-evaluation of constipation by the respondents and objective criteria was higher for negatives (95.8%) than for positives (38.0%). Conclusion: The prevalence of constipation is relatively high in the urban population of Hangzhou. A good lifestyle and diet can significantly reduce its incidence. There should be more emphasis on preventive education, promotion of prevention of constipation, and advocacy for a healthy lifestyle.

[Detection and analysis of intestinal flora diversity in patients with complex anal fistula].

Objective: To explore the possibility that the intestinal flora profile in complex anal fistula patients is different to that of healthy controls. This was assessed by sequencing of 16S rDNA in fecal samples from cohorts representing these populations. Methods: Fecal samples were collected from 30 complex anal fistula patients and 30 matched healthy controls. Patients were included if they met the diagnostic criteria of cryptoglandular anal fistula and had exhibited symptoms for more than 3 months. Complex anal fistula is diagnosed under the following circumstances: if the fistula in question spans 2/3 or more of the diameter of the anal sphincter; if there are more than two external orifices or fistula tracks; or if recurrence is observed after previous anal fistula surgery. Patients were excluded if there were comorbities including inflammatory bowel disease (as assessed by colonoscopy), chronic diarrhea, chronic constipation, diabetes, gastrointestinal malignancies, liver/ kidney dysfunction, or cognitive impairment. Patients whose anal fistulas were caused by Crohn's disease, trauma, special infections (such as actinomycosis and tuberculosis) were also excluded, as were those who had used antibiotics, prebiotics, or probiotics that may affect intestinal microecology in the month prior to the study. Total bacterial genomic DNA was extracted by PCR amplification of the V4 hypervariable region of the 16S rRNA sequences. High-throughput sequencing and data analysis were performed on the Illumina Miseq platform. Finally, operational taxonomic unit (OTU) clustering, alpha diversity and LEfSE data analysis were carried out. The larger the Chao or ACE index is, the higher the species abundance of the microflora is expected to be. Similarly, a smaller value for the Simpson index or a larger value for the Shannon index indicates greater microflora diversity. There was no statistically significant difference in gender, age, body mass index (BMI), drinking history, or smoking history between the two groups (P>0.05), indicating that they were comparable. Results: The α-diversity analysis including ACE, Chao, Shannon and Simpson indexes indicated a richer diversity of intestinal microflora in complex anal fistula patients than in healthy controls. In both patients and controls, OUT cluster analysis demonstrated that 93.4%±32.0% and 87.4%±41.2% of sequences were from Firmicutes and Bacteroidetes spp., respectively. On a genus level, samples from anal fistula patients showed a greater abundance of Prevotella spp. (4.9%±7.4% vs. 0.1%±1.1%, P<0.001), Megamonas (3.9%±8.2% vs. 0.5%±4.2%, P<0.05) and Lachnospira (2.6%±5.7% vs. 0.1%±3.4%, P<0.05), while showing a lesser abundance of Proteobacteria spp. (0.02%±4.2% vs. 9.3%±14.4%, P<0.01), Enterococcus (0.02%±2.3% vs. 9.3%±19.6%, P<0.05), Bacteroides (24.7%±9.9% vs. 29.8%±9.1%, P<0.05) and Klebsiella (0.4%±4.2% vs. 3.9%±7.3%, P<0.05) compared with healthy controls. Intestinal flora diversity in the complex anal fistula group was richer than in controls, as indicated by a higher ACE index (293.30±44.00 vs. 218.75±33.83, t=102.069, P<0.001), a higher Chao index (318.40±41.99 vs. 250.00±46.38, t=77.818, P=0.028), a higher Shannon index (3.36±0.29 vs. 2.43±0.34, t=9.657, P=0.001), and a lower Simpson index (0.103±0.013 vs. 0.131±0.013, t=5.551, P=0.046). LDA effect size analysis suggests that the main strains of Veillonellaceae, Selenemondales and Negativicutes, which all belong to the phylum Firmicutes, have the greatest influence on the above difference (LDA>4). Conclusions: The diversity of intestinal flora in patients with complex anal fistula is greater than in healthy controls, suggesting that these bacteria or their metabolites may be involved in the occurrence and development of anal fistulas.

[Expression and functional SNP loci screen of ATM from coal worker's pneumoconiosis].

Objective: To detect of gene expression and genotype of the ataxia telangiectasia mutated (ATM) from coal workers' pneumoconiosis (CWP) , It is explored whether CWP is related to ATM gene. Methods: In October 2020, the relevant information of 264 subjects who received physical examination or medical treatment in the Department of occupational diseases of Guiyang public health treatment center from January 2019 to September 2020 was collected. Through the occupational health examination, 67 healthy people with no history of exposure to occupational hazards were selected as the healthy control group; The coal miners with more than 10 years of coal dust exposure history and small shadow in the lung but not up to the diagnostic criteria were the dust exposure control group, a total of 66 people; The patients with the same history of coal dust exposure and confirmed stage I were coal worker's pneumoconiosis stage I group, a total of 131 people. The expression of ATM was detected by QRT PCR. ATM rs189037 and rs1801516 were genotyped by massarray. Results: There was significant difference in the expression of ATM among the groups (P<0.05) ; Compared with the healthy control group, the expression of ATM in the dust exposed control group was significantly increased (P<0.05) . With the occurrence and development of CWP, the GG of rs189037 wild type decreased, the GA of mutant heterozygote and AA of homozygote increased, but the difference was not statistically significant (P>0.05) ; Rs1801516 wild type GG and mutant heterozygote GA had no significant changes (P>0.05) . There were significant differences in age, neutrophils and basophils among rs189037 groups (all P<0.05) . There were no significant differences in blood pressure, eosinophils, lymphocytes, monocytes, smoking and drinking history among rs189037 groups (all P>0.05) . Compared with wild-type GG, the or of mutant heterozygotes and homozygotes increased, but the differences were not statistically significant (P>0.05) . Conclusion: ATM gene may be one of the early activation genes of CWP and rs189037 may be the functional loci which affects gene expression. ATM gene is related to inflammatory response, Neutrophils and basophils have an impact on the development of CWP.

[A comparison of colonoscopy - assisted transanal minimally invasive surgery via glove port and endoscopic submucosal dissection in the treatment of early rectal tumors].

Objective: To explore the safety and feasibility of colonoscopy - assisted transanal minimally invasive surgery via glove port (CA-TAMIS-GP) in the treatment of early rectal tumors. Methods: A total of 67 patients evaluated as early rectal tumors (adenoma limited within mucosal layer) with diameter ≤4.0 cm at Department of Anal-Colorectal Surgery, Hangzhou Third People's Hospital from July 2013 to March 2017 were prospectively enrolled in the study. Benign tumors were diagnosed by preoperative imaging in all the patients with the distance to anal edge of 4 to 20 cm. Patients were randomly divided into treatment group and the control group according to the random number table. The treatment group (n=32) underwent CA-TAMIS-GP, including 19 males and 13 females with mean age of (55.6±11.2) years and mean tumor size of (3.3±0.4) cm, while the control group (n=35) underwent endoscopic submucosal dissection (ESD, control group), including 20 males and 15 females with mean age of (52.9±12.3) years and mean tumor size of (3.4±0.5) cm. Differences of baseline data between two groups were not significant (all P>0.05). The specific method of CA-TAMIS-GP was as follows: a surgical rubber glove sleeve (No.6) was passed through the anal device; the glove was fixed at the anvil device; after fully expanding the anus, the anal sac was placed into the anus with the fingers outside; then, the cuff and the anal sac were sutured and fixed to the perianal; a well-tight glove path was established; the ultrasonic scalpel, grasper and the colonoscopy lens connected to the host platform and the electric negative pressure suction were inserted into the three finger sleeves respectively and fixed by rubber band or silk thread; the laparoscopic instruments such as the grasper and the ultrasonic scalpel were used for pulling, grasping, cutting, electrocoagulation, suturing and other operations to complete the resection of rectal lesions. Efficacy, postoperative complication and operative cost, etc. between two groups were compared using the student's t test, chi-square tests, and Fisher's exact test. Results: Operations of two groups were completed successfully without conversion to laparotomy. Histopathologic examination showed all specimens had negative margins with the surgical resection of the layer to the submucosa, and showed no significant differences between two groups (P>0.05). Compared to the control group, the operation time was shorter [(49.5±14.6) minutes vs.(66.1±17.6) minutes, t=-4.235, P<0.001], and the intraoperative hemorrhage was less [(4.2±1.6) ml vs. (6.2±2.1) ml, t=-4.349, P<0.001] in the treatment group with significant differences. In the treatment group, 6 patients had mild anal pain or discomfort after operation, and 1 patient in the control group showed anal foreign body sensation. The difference was statistically significant [18.8% (6/32) vs. 2.9% (1/35), P=0.048]. The incidence of postoperative hematochezia in the treatment group was lower than that in the control group [9.4% (3/32) vs. 20.0% (7/35), P=0.310] without significant difference. The cost of consumables in the treatment group was (1586.9±204.4) yuan, which was lower than (7694.4±1123.2) yuan in control group, and the difference was statistically significant (t=-30.880, P<0.001). All the patients were followed up for 6 to 36 months after operation, and no recurrence or long-term complication occurred in the treatment group, while 1 case developed local recurrence in the control group. Conclusion: CA-TAMIS-GP is a safe and effective method for early rectal tumors with simple and economical characteristics, which broadens the application of colonoscopy.

[The status quo of amphetamine type stimulants use among residents aged 15-64 in a border city of Yunnan province, 2014].

Objective: To understand status of amphetaminetype stimulants (ATS) use among residents aged 15-64 in a border city of Yunnan province. Methods: Using the stratified cluster random sampling method, a total of 3 130 residents were recruited through both anonymous questionnaire and interview, regarding their health-related behaviour and ATS use. Results: Among 3 130 residents aged 15-64 years in this city, the overall prevalence rates of ATS use were 4.0% (126/3 130) in the lifetime and 2.6% (82/3 130) in the past one year, while the prevalence of ATS use disorder in the past year was 2.3% (73/3 130). The prevalence rates of lifetime ATS use and in the past year were 7.5% (108/1 443) and 5.1% (73/1 443) in the high epidemic area, 7.3% (122/1 682) and 4.8%(80/1 682) in males, 5.2% (118/2 260) and 3.4% (77/2 260) in 18-45 age group, 4.6%(63/1 361) and 3.4% (46/1 361) in the ones having had elementary school education, 10.3% (50/487) and 8.6% (42/487) in unmarried group, 17.1% (19/111) and 12.6% (14/111) in either divorced, widowed or separated group, 4.8% (108/2 256) and 3.2% (72/2 256) in the farmers group, 6.0% (99/1 643) and 4.4% (73/1 643) in the non-religious groups (neither Buddhism nor Christianity), 15.2% (97/639) and 11.9% (76/639) in groups with negative hobbies which were 1.3% (29/2 314) and 0.3%(6/2 314) higher than the ones with positive hobbies, 8.5% (84/992) and 6.3% (62/992) in the ones negatively managing the stress which were 2.0% (42/2 138) and 0.9% (20/2 138) higher than the ones that treating the stress in a positive way. Conclusions: The prevalence rates of ATS use and ATS use disorder in the past one year appeared high among those residents aged 15-64 years in the border city of Yunnan province. Prevention and intervention programs should be carried out for the target groups as soon as possible.

LncRNALUADT1 is overexpressed in colorectal cancer and its expression level is related to clinicopathology.

OBJECTIVE: LncRNAs participate in the proliferation, apoptosis, and invasion of colorectal cancer. We aimed at investigating the uncovered effect of lncRNALUADT1 on colorectal cancer. PATIENTS AND METHODS: The relative expression level of lncRNALUADT1 in tumor specimen was tested by Real-time quantitative PCR. The association of lncRNALUADT1 with clinical pathological data was analyzed by univariate, multivariate Cox and Kaplan-Meier curve. RESULTS: LncRNALUADT1 expression was up-regulated in colorectal cancer, and correlated with tumor size, metastasis, and TNM staging. Both univariate analysis and multivariate test indicated that lncRNALUADT1 high expression, TNM staging, and lymph node metastasis were closely related. Moreover, high expression of lncRNALUADT1 suggested the poor overall survival of patients. CONCLUSIONS: LncRNA LUADT1 might contribute to the development of colorectal cancer.

Linker length affects expression and bioactivity of the onconase fusion protein in Pichia pastoris.

The aim of this study was to analyze the effect of linker length on the expression and biological activity of recombinant protein onconase (ONC) in fusion with human serum albumin (HSA) in Pichia pastoris. Four flexible linkers with different lengths namely Linker L0, L1: (GGGGS)1, L2: (GGGGS)2, and L3:(GGGGS)3 were inserted into the fusion gene and referred to as HSA-n-ONC, where N = 0, 5, 10, or 15. The sequence of the fusion gene HSA-ONC was designed based on the GC content and codon bias in P. pastoris; the signal peptide of albumin was used as the secretion signal. Gene sequences coding for the fusion protein with different linkers were inserted into pPICZα-A to form recombinant plasmids pPICZα-A/HSA-n-ONC, which were then transformed into P. pastoris X-33 for protein expression. Ideal conditions for expression of the fusion proteins were optimized at a small scale, using shake flasks before proceeding to mass production in 10-L fermenters. The recombinant fusion proteins were purified by aqueous two-phase extraction coupled with DEAE anion exchange chromatography, and their cytotoxic effect on the tumor cell was evaluated by the sulforhodamine B assay. The results showed that the expressed amount of fusion proteins had no significant relationship with the length of different linkers and rHSA-0-ONC had no cytotoxic effect on the tumor cells. While rHSA-5-ONC and rHSA-10-ONC had a weak cytotoxic effect, rHSA-15-ONC could kill various tumor cells in vitro. In summary, the biological activity of the fusion protein gradually improved with increasing length of the linker.

Hepatitis B virions isolated with antibodies to the pre-S1 domain reveal occult viremia by PCR in Alaska Native HBV carriers who have seroconverted.

BACKGROUND: Occult viremia occurring before the appearance of HBsAg or after the disappearance of HBsAg is detectable by gene amplification technologies whose efficiency depends on nucleic acid preparation. STUDY DESIGN AND METHODS: To isolate HBV DNA from viremic plasma, immunoaffinity capture (IAC) of intact HBV with biotinylated pre-S1 antibodies coupled to streptavidin-coated magnetic beads was evaluated. IAC was compared with a silica-gel method (Qiagen [QSG]) and its two modifications wherein the samples were heated with lysis buffer at 60(o)C for 10 minutes (QSG-60) or at 58 degrees C for 60 minutes with proteinase-K (QSG-PK). Each HBV DNA sample was tested by heminested PCR amplification of the HBV gene sequences. A total of 36 coded serum samples were tested, including three HBsAg-positive controls and 33 former chronic HBV carriers who had seroconverted (developed antibody to HBsAg [anti-HBs]). Commercially available seroconversion panels (PHM 907, 911, and 922) were similarly tested for window-period viremia. RESULTS: In the 33 former chronic HBV carriers who had seroconverted, IAC revealed HBV DNA in 17 samples, whereas it was revealed in only 11 samples by QSG-PK (p = 0.031), 10 by QSG-60 (p = 0.016), and 9 by QSG (p = 0.0078). However, HBV DNA was not amplified from the 17 samples at 1-in-10 dilutions; thus, they were considered to have low-level viremia. IAC revealed HBV DNA as early as or earlier than the other methods in PHM 907, 911, and 922 panels. CONCLUSION: IAC is apparently an optimal method of sample preparation for amplification of HBV DNA in patients in the pre-HBsAg window period, and for detecting low-level viremia persistent in several individuals who were former chronic HBV carriers who had seroconverted (developed anti-HBs).

Properties of cyanovirin-N (CV-N): inactivation of HIV-1 by sessile cyanovirin-N (sCV-N).

Cyanovirin-N (CV-N) is a novel anti-HIV protein isolated and characterized from a cyanobacterium Nostoc ellipsosporum. CV-N protein is a single 101 amino acid chain containing two intrachain disulphide bonds and considerable internal sequence duplication, but no significant homology to previously described proteins or to the transcription products of known nucleotide sequences. In solution, CV-N exists largely as a beta-sheet protein with internal two-fold pseudosymmetry. CV-N irreversibly inactivates diverse laboratory strains, primary isolates and clades of HIV-1, as well as strains of HIV-2 and simian immunodeficiency virus (SIV). CV-N binds with extremely high affinity to highly conserved binding site(s) on the viral envelope glycoprotein gp120, preventing virus-to-cell fusion, viral entry and infection of cells. The CV-N binding site appears to overlap, but is not identical with, the unique carbohydrate-dependent epitope 2G12, and may lie predominantly within an immunologically "silent" region of gp120. CV-N is undergoing preclinical development for topical anti-HIV prophylactic (e.g., microbicidal) applications to prevent sexual transmission of HIV. Since CV-N may be immunogenic in humans, methods for using CV-N for ex vivo inactivation of HIV in blood, plasma, or putative vaccines preferably would allow for its exclusion from biologicals for parenteral use. To explore this concept we biotinylated CV-N (bCV-N) and coupled it to streptavidin coated magnetic beads to provide a product which we termed sessile CV-N (sCV-N). When reacted with a laboratory strain and a primary isolate of HIV- 1, the sCV-N completely inactivated 100 TCID50 of the virus. However RT-PCR of the viral extracts indicated that only a fraction of the virus was removed by the sCV-N, leaving behind a relatively larger fraction of non-infectious virus in the supernatant which we designated as replication incompetent virions (RIV). It would be worthwhile investigating the role of RIV as a putative HIV vaccine.

Volume reflection holographic confocal imaging.

The Denisyuk volume reflection hologram is produced with spatially incoherent light to form an image-plane hologram. The image formed in readout combines the properties of volume holography and confocal image formation.

Hemodynamic effects of brief pulmonary hemostasis.

We determined the hemodynamic effects of brief hemostasis in isolated blood-perfused rat lungs held at constant inflation pressure. Three periods of hemostasis, each lasting 20 min, alternated with equal durations of perfusion. Pulmonary vascular resistance (PVR), calculated as the ratio of the pulmonary arterial- (Ppa) left atrial (Pla) pressure difference to flow, was determined at baseline and after the third hemostasis period. Hemostasis increased PVR by 127% above baseline at constant Ppa and Pla (n = 8; P < 0.05) and by 71% at constant Pla and constant flow (n = 16; P < 0.05). The PVR increases were significantly attenuated by platelet (n = 5) or red blood cell (n = 4) removal from the lung perfusion, mechanical ventilation (n = 5), indomethacin (2 X 10(-6) M; n = 5), papaverine (10(-5) M; n = 4), and the thromboxane blocker SQ-29548 (7.8 X 10(-5) M; n = 4). By micropuncture we determined that the posthemostasis increase of the total pressure drop was 66% due to an increase of the venous pressure drop (P < 0.05). Isogravimetric pressure and lung water were not significantly altered by hemostasis. We conclude that brief hemostasis in the rat lung induces platelet- and red blood cell-dependent increase of vascular resistance. This effect may be attributable to hemostasis-induced platelet activation leading to thromboxane release.

Sonographic patterns of ribs with tumor involvement.

We analyzed the sonographic appearance of the ribs when there was tumor involvement in 16 patients (12 men and 4 women). The tumors included 11 carcinomas, 3 nonepithelial malignancies and 2 benign tumors. Three sonographic patterns on the cross section of abnormal ribs were identified. Pattern I was an eccentrical hyperechoic plate-like shadow inside a hypoechoic tumor. Pattern II was a round hyperechoic shadow or ring in the center of a hypoechoic tumor. Pattern III was a hypoechoic tumor only. The Pancoast tumor manifested pattern I. Metastatic cancer, mostly squamous cell carcinoma, manifested pattern II, and both malignant and benign tumors could manifest pattern III. In sonography, the margin and shape of the tumors, the change in the acoustic shadow of the rib and the pleural line did not differentiate the malignant tumors from benign tumors. When we routinely took a biopsy of these tumors under sonar guidance, the diagnostic yield was 100% without any complications. We conclude that the rib should be evaluated in chest sonography, and that pattern recognition and biopsy under sonar guidance are most useful.

Design of a general optical interconnection system with phase-only masks.

A general optical interconnection system with phase-only masks is proposed; it is a high-efficiency system. The optical general transform theory is modified for the design. Results show that an arbitrary one-to-one interconnection pattern between an array of N sources and N detectors can be achieved by two phase-only masks in which the number of sampling points is N(2).