An Exo III-powered closed-loop DNA circuit architecture for biosensing/imaging.

With their regulated Boolean logic operations in vitro and in vivo, DNA logic circuits have shown great promise for target recognition and disease diagnosis. However, significant obstacles must be overcome to improve their operational efficiency and broaden their range of applications. In this study, we propose an Exo III-powered closed-loop DNA circuit (ECDC) architecture that integrates four highly efficient AND logic gates. The ECDC utilizes Exo III as the sole enzyme-activated actuator, simplifying the circuit design and ensuring optimal performance. Moreover, the use of Exo III enables a self-feedback (autocatalytic) mechanism in the dynamic switching between AND logic gates within this circulating logic circuit. After validating the signal flow and examining the impact of each AND logic gate on the regulation of the circuit, we demonstrate the intelligent determination of miR-21 using the carefully designed ECDC architecture in vitro. The proposed ECDC exhibits a linear detection range for miR-21 from 0 to 300 nM, with a limit of detection (LOD) of approximately 0.01 nM, surpassing most reported methods. It also shows excellent selectivity for miR-21 detection and holds potential for identifying and imaging live cancer cells. This study presents a practical and efficient strategy for monitoring various nucleic acid-based biomarkers in vitro and in vivo through specific sequence modifications, offering significant potential for early cancer diagnosis, bioanalysis, and prognostic clinical applications.

Clinical evaluation of a multitarget fecal immunochemical test-sDNA test for colorectal cancer screening in a high-risk population: a prospective, multicenter clinical study.

Colorectal cancer (CRC) is a major malignancy threatening the health of people in China and screening could be effective for preventing the occurrence and reducing the mortality of CRC. We conducted a multicenter, prospective clinical study which recruited 4,245 high-risk CRC individuals defined as having positive risk-adapted scores or fecal immunochemical test (FIT) results, to evaluate the clinical performance of the multitarget fecal immunochemical and stool DNA (FIT-sDNA) test for CRC screening. Each participant was asked to provide a stool sample prior to bowel preparation, and FIT-sDNA test and FIT were performed independently of colonoscopy. We found that 186 (4.4%) were confirmed to have CRC, and 375 (8.8%) had advanced precancerous neoplasia among the high CRC risk individuals. The sensitivity of detecting CRC for FIT-sDNA test was 91.9% (95% CI, 86.8-95.3), compared with 62.4% (95% CI, 54.9-69.3) for FIT (P < 0.001). The sensitivity for detecting advanced precancerous neoplasia was 63.5% (95% CI, 58.3-68.3) for FIT-sDNA test, compared with 30.9% (95% CI, 26.3-35.6) for FIT (P < 0.001). Multitarget FIT-sDNA test detected more colorectal advanced neoplasia than FIT. Overall, these findings indicated that in areas with limited colonoscopy resources, FIT-sDNA test could be a promising further risk triaging modality to select patients for colonoscopy in CRC screening.

A comparison of the efficacy and safety of natural orifice specimen extraction and conventional laparoscopic surgery in patients with sigmoid colon/high rectal cancer.

OBJECTIVE: This study aimed to investigate the efficacy and safety of natural orifice specimen extraction surgery (NOSES) compared to conventional laparoscopic radical resection in the treatment of patients with sigmoid colon/high rectal cancer. METHODS: The control group (n = 62) underwent traditional laparoscopic radical resection, and the observation group (n = 62) underwent transanal NOSES laparoscopic radical resection. The operation length; amount of bleeding; number of lymph node dissections and days of hospitalization after surgery; visual pain scores on the first and third days after surgery; first leaving bed, anal exhaust, eating a liquid diet, and effective sleep times; and the postoperative complications (abdominal or incision infection or anastomotic fistula) of the two groups of patients were compared and analyzed. RESULTS: The effective sleep time of the observation group on the first day after the operation was 12.3 ± 2.9 h, which was longer than that of the control group (10.6 ± 3.2 h), and the difference was statistically significant (p < 0.001). The pain degree of the two groups on the third day after the operation was lower than that on the first day, and the pain score of the observation group was lower than that of the control group (2.0 ± 1.0 vs. 3.2 ± 1.2, p < 0.001). The postoperative hospital stay in the observation group was significantly shorter than that in the control group (9.7 ± 2.3 vs. 11.2 ± 2.6, p < 0.001). The incidence of postoperative complications in the observation group was significantly lower than that in the control group (3.2% vs. 12.9%, p = 0.048). In addition, it was found that the first leaving bed, anal exhaust and liquid diet times in the observation group were significantly shorter than those in the control group (p < 0.001). CONCLUSION: Laparoscopic radical resection NOSES in patients with sigmoid colon cancer or high rectal cancer leads to lower postoperative pain and longer sleep time than in patients who undergo traditional laparoscopic radical surgery. The complication rate of this procedure is low, and the curative effect is safe and positive.

The Pharmacokinetics and Pharmacodynamics of A Novel Recombinant Activated Human Factor VII, GEN-0828, in Hemophilia B Mice.

GEN-0828, a proposed clinical candidate for hemophilia and trauma hemorrhage treatment, is a novel recombinant activated human factor VII (rFVIIa). The purpose of this paper is to compare the pharmacokinetics and pharmacodynamics of GEN-0828 in hemophilia B mice with those of NovoSeven®, the only marketed rFVIIa product worldwide., GEN-0828 and NovoSeven® showed similar affinity bioactivity to recombinant tissue factor (rTF) in vitro. Pharmacodynamics data indicated a generally similar hemostatic efficacy (ED50) of GEN-0828 (10.91 KIU·kg-1) and NovoSeven® (18.91 KIU·kg-1) at the doses studied in hemophilia B mice, while GEN-0828 represented a lower initial effective dosage compared with that of NovoSeven® in terms of both blood loss and APTT. GEN-0828 exhibited linear pharmacokinetic profiles in hemophilia B mice at the 30-338 KIU·kg-1 dose range, the comparative pharmacokinetic study with NovoSeven® indicated better characteristics than NovoSeven® in terms of the appropriate higher maximal concentration (Cmax) and area under the plasma concentration-time curve (AUClast) and longer mean residence time (MRT). In conclusion, GEN-0828 was a promising new type of rFVIIa compound with favourable pharmacokinetic and pharmacodynamic profiles.

Evaluation of the metabolism of PEP06, an endostatin-RGDRGD 30-amino-acid polypeptide and a promising novel drug for targeting tumor cells.

PEP06 is a novel endostatin-Arg-Gly-Asp-Arg-Gly-Asp (RGDRGD) 30-amino-acid polypeptide featuring a terminally fused RGDRGD hexapeptide at the N terminus. The active endostatin fragment of PEP06 directly targets tumor cells and exerts an antitumoral effect. However, little is known about the kinetics and degradation products of PEP06 in vitro or in vivo. In this study, we investigated the in vitro metabolic stability of PEP06 after it was incubated with living cells obtained from animals of different species; we further identified the degradation characteristics of its cleavage products. PEP06 underwent rapid enzymatic degradation in multiple types of living cells, and the liver, kidney, and blood play important roles in the metabolism and clearance of the peptides resulting from the molecular degradation of PEP06. We identified metabolites of PEP06 using full-scan mass spectrometry (MS) and tandem MS (MS2), wherein 43 metabolites were characterized and identified as the degradation metabolites from the parent peptide, formed by successive losses of amino acids. The metabolites were C and N terminal truncated products of PEP06. The structures of 11 metabolites (M6, M7, M16, M17, M21, M25, M33, M34, M39, M40, and M42) were further confirmed by comparing the retention times of similar full MS spectrum and MS2 spectrum information with reference standards for the synthesized metabolites. We have demonstrated the metabolic stability of PEP06 in vitro and identified a series of potentially bioactive downstream metabolites of PEP06, which can support further drug research.

Comparative analysis of modified Bacon operation and double stapler operation in the treatment of rectal cancer.

OBJECTIVE: To investigate the efficacy of modified Bacon operation and double stapler operation in treating lower rectal cancer. METHODS: A total of 73 patients with lower rectal cancer admitted to the general surgery department of our hospital from March 2016 to March 2017 were selected for the study. According to different operation methods, the patients were divided into an observation group and a control group. Modified Bacon operation was used in the observation group (39 cases), while double stapler operation was adopted for the control group (34 cases). The intraoperative and postoperative conditions and 3-year survival rates were observed. RESULTS: The bleeding volume and anal exhaust time in the observation group were better than those in the control group, the difference being statistically significant (P<0.05). No significant difference was found between the two groups in terms of the operation time, abdominal drainage volume, anal extubation time, or the number of lymph node dissection (P>0.05). The anal function of the two groups was evaluated one month and six months after operation respectively, and no significant difference was found between the function of the two groups (P>0.05). There was no significant difference in postoperative complications, local recurrence, and distant metastasis between the two groups (P>0.05). The 3-year survival rate was 89.74% in the observation group and 91.18% in the control group. Our analysis found no significant difference in the cumulative survival rates between the two groups (P>0.05). CONCLUSION: Modified Bacon operation and double stapler operation are effective treatment for lower rectal cancer, and the survival benefit of patients is obvious. The methods are worthy of clinical promotion.

An optimized LC-MS/MS method for determination of HYNIC-3PRGD2, a new promising imaging agent for tumor targeting, in rat plasma and its application.

HYNIC-3PRGD2 is used to prepare a new 99mTc-radiolabeled tracer. HYNIC-3PRGD2, which has a high binding affinity for the integrin αvß3 due to its special structure, has become a promising tumor imaging agent for diagnosis and monitor of the clinical response to therapeutic effects of anti-tumor agents. Here, we developed and validated a method for determination of HYNIC-3PRGD2 concentration in rat plasma using ultra-high performance liquid chromatography-tandem mass spectrometry system. Following sample extraction by methanol precipitation, satisfactory separation through chromatography was achieved on an hydrophilic reverse-phase C18 column AQ (2.1 mm × 100 mm, 2.7 µm) at a flow rate of 0.2 mL·min-1 with an gradient elution using mobile phase consisting of ultrapure water and acetonitrile fortified with 0.1% formic acid respectively. The calibration curve was developed over a linear range of 3.125-100 ng·mL-1 with the lower limit of quantification of 3.125 ng·mL-1. The HYNIC-3PRGD2 and its internal standard c(RGDfK)(RK5) were detected and quantified with the multiple reaction monitoring (MRM) mode on a triple-quadrupole tandem mass spectrometer. This method was successfully validated and applied for pharmacokinetic evaluation of HYNIC-3PRGD2 during pre-clinical experiments.

[Clinicopathological characteristics and prognosis analysis of colorectal synchronous multiple primary cancer].

OBJECTIVE: To investigate the clinicopathological features and prognosis of colorectal synchronous multiple primary cancer(SMPC). METHODS: From January 2008 to June 2011, 51 patients diagnosed with colorectal SMPC underwent surgery at Department of General Surgery of Peking University First Hospital. Their clinicopathological features, diagnosis, treatment and prognosis were summarized and analyzed. SMPC was diagnosed according to the following criteria: each tumor must have a definite pathologic picture of malignancy; metastasis or recurrence from another colorectal cancer was excluded; tumors must be distinctly separated by at least 5 cm of all intact bowel wall from each other; SMPC has abnormal cells between tumor and normal mucosa and abnormal gland of transitional zone; each cancer is infiltrating carcinoma except the carcinoma in situ; all the cancers are detected at the same time or within 6 months. Multiple primary colorectal cancer originated from familial colonic polyposis or ulcerative colitis was excluded. RESULTS: These 51 colorectal SMPC patients accounted for 3.5% of 1 452 colorectal cancer patients in the same period at our hospital, with 32 males and 19 females, and mean age of (63±13)(29 to 82) years. Of 51 cases, 46(90.2%) had 2 original carcinoma, 3(5.9%) had 3 original carcinoma and 2(3.9%) had 4 carcinoma; 23(45.1%) complicated with colon polyps, 4(7.8%) complicated with malignancy outside the colorectum. In TNM staging, 7(13.7%), 15(29.4%), 24(47.1%) and 5(9.8%) patients were stage I(, II(, III( and IIII( respectively. Among 51 patients undergoing surgery by different procedures, 16 were subtotal colon resection, 8 were extended right colon resection, 5 were extended left hemicolon resection, 8 were right hemicolon resection plus Dixon procedure, 10 were Dixon, and 4 were right hemicolon resection plus sigmoid colon resection. Adjuvant chemotherapy and support treatment were given according to the condition after operation. A total of 105 tumors were found, including 25(23.8%) tumors in sigmoid colon, 24(22.9%) in rectum, 22(21.0%) in ascending colon and 4 in organs outside the colorectum. Tubular adenocarcinoma (86/105, 81.9%) was the main pathological type in these colorectal SMPC patients. During the follow-up of median 43.5 months, 10 cases presented local recurrence and 6 cases had liver metastasis. Multivariable analysis showed that ≤65 years old (OR=22.757, 95%CI: 1.562-331.543, P=0.002),undifferentiated carcinoma or mucous adenocarcinoma (OR=27.174, 95%CI: 2.834-260.512, P=0.004), stage III(-IIII( (OR=29.626, 95%CI: 3.216-272.884, P=0.003) were independent risk factors of postoperative 5-year recurrence and metastasis, but the number of SMPC lesions and the surgical method were not associated with postoperative 5-year recurrence and metastasis (P=0.564, P=0.513). The 3-year and 5-year survival rates of colorectal SMPC patients were 76.5% and 64.7%. CONCLUSION: Two-original carcinoma is the most common in colorectal SMPC patients, which mainly distributes in sigmoid colon and rectum. Postoperative monitoring should be strengthened for those patients with younger age, poor pathological types and advanced staging to prevent recurrence and metastasis.

Retrospective series of subtotal colonic bypass and antiperistaltic cecoproctostomy for the treatment of slow-transit constipation.

PURPOSE: Some popular surgical methods with big trauma to treat slow-transit constipation (STC) at present are far from satisfactory. This work is aimed to evaluate the clinical and functional outcomes of a redesigned procedure of subtotal colonic bypass and antiperistaltic cecoproctostomy (SCBAC) for the treatment of STC. METHODS: The clinical data of 18 patients with severe idiopathic STC treated with SCBAC between September 2003 and September 2008 were retrospectively analyzed. The last eight patients accepted SCBAC were assessed by short form-36 (SF-36) scores translated into Chinese to inquire upon the quality of life before and 6 months after operation. RESULTS: There were no procedure-related deaths in this series (mortality, 0%) or any serious complication. The average follow-up period was 17 months (range, 6-60 months). There was no intestinal occlusion due to adhesions that required surgery in all these patients. Scores for all SF-36 subscales were significantly higher for patients after operation than for before operation. A high number of patients (83%) expressed a willingness to repeat the procedure given the same preoperative conditions. CONCLUSION: Our clinical practice demonstrates that the procedure of SCBAC is effective for the treatment of severe idiopathic STC in some cases. Due to its mild trauma and less operating time, SCBAC is almost well-tolerated and compatible in all age groups, making it worthy of further practice and investigation.

A modified Delorme's operation for the treatment of rectal mucosal prolapse.

AIMS: Many procedures have been described for the treatment of rectal internal mucosal prolapse (RMP), but the therapeutic effect is questionable. This work is to evaluate clinical and functional outcome of a modified Delorme's operation-trans-anal purse-string sutures for rectal mucosa and submucosa (TAS) for the treatment of RMP. METHODS: The clinical data of 58 patients with rectal mucosal prolapse treated with rectal mucosa and submucosal tissue (TAS) between June 2004 and June 2008 were analyzed retrospectively. RESULTS: No patient died. Satisfaction with surgery was high in 48 cases (82.8%), moderate in seven (12.1%), and low in three (5.17%). Prolapse relapse rate was 5.17%. Anal tenesmus and urge to defecate resolved in 52 (89.7%) patients. Constipation improved in 25 of 28 (89.3%) previously constipated patients. No surgery-associated constipation occurred. The mean operative time was 31 (range 22-46) min. Mean hospital stay was 3 days (range 2-6). Mean patient follow-up was 32 months (range 12-60). CONCLUSIONS: From our data, TAS for the treatment of RMP showed encouraging results with little complications and an acceptable relapse rate. This economical procedure induces only mild trauma and is easy to perform, making it worthy of further practice and investigation.

An effective perineal procedure for the management of full-thickness rectal prolapse.

AIMS: To evaluate the clinical and functional outcome of transanal purse-string sutures for rectal mucosa and submucosa plus perianal suture (TAS-PAS) for the management of full-thickness rectal prolapse. METHODS: The clinical data of 62 patients with full-thickness rectal prolapse treated with TAS-PAS between March 2000 and March 2008 were analyzed retrospectively. RESULTS: No patient died. Satisfaction with surgery was high in 50 cases (80.6%), moderate in 9 cases (14.5%), and low in 3 cases (4.84%). Prolapse relapse rate was 4.84%. Anal continence improved in 82.6% of patients, constipation improved in 69.2%, and anal tenesmus in 86.7%. No surgery-associated constipation occurred. The mean operative time was 52 min (range 40-80). Mean hospital stay was 4 days (range 3-7). Mean patient follow-up was 17 months (range 4-36). CONCLUSIONS: From our data, TAS-PAS for the management of full-thickness rectal prolapse showed encouraging results with little complications and an acceptable relapse rate. This procedure induces only mild trauma and is easy to perform making it worthy of further practice and investigation.

[Association of soluble human leukocyte antigen DQB1 with recurrent spontaneous abortion].

OBJECTIVE: To investigate the relationship between the recurrent spontaneous abortion and soluble human leukocyte antigen (HLA) DQB1 gene polymorphism. METHODS: Sixty one cases with recurrent spontaneous abortion were included in study group. Of them, 31 were anticardiolipin antibody (ACA) positive, while 30 were negative. Twenty eight cases were with early abortion, 22 were with late abortion, and 11 were with both early and late abortions. Thirty six women without abnormal pregnancy history were recruited as control group. HLA-DQB1 genotyping was carried out by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP). RESULTS: The frequency of DQB1*0201allele in the study group (16.4%, 20/122) was higher than that in the control group (8.3%, 6/72), but the difference was not statistically significant (P > 0.05). The frequency of 57 site with non Asp in the study group was significantly higher than that in the control group (P < 0.05). The frequencies of both DQB1*0201 and *0303 were significantly different among early abortion, late abortion and both early and late abortion groups (P < 0.05 and P < 0.05). The frequency of DQB1*0201 was significantly higher in the early abortion group than that in the late abortion group (P < 0.01). The frequency of DQB1*0303 was significantly increased in the late abortion group when compared to that in the early group (P < 0.01). The frequency of DQB1*0303 was found to be significantly higher in patients with ACA positive than that in ACA negative group (P < 0.05). CONCLUSION: The 57 site with non Asp may be a gene related to recurrent spontaneous abortions. DQB1*0201 may be correlated to the early abortion, while DQB1*0303 correlated to the late abortion and abortion with ACA positive.