[Antitumor effects of redox-responsive nanoparticles containing platinum（⠣）in ovarian cancer].

Objectives: To explore the antitumor effects of redox-responsive nanoparticles containing platinum(Ⅳ)-NP@Pt(Ⅳ) in ovarian cancer. Methods: Redox-responsive polymer carriers were synthesized. Polymer carriers and platinum(Ⅳ)-Pt(Ⅳ) can self-assemble into NP@Pt(Ⅳ). Inductively coupled plasma mass spectrometry was performed to detect the platinum release from NP@Pt(Ⅳ) in reducing environment and the platinum content in ovarian cancer cells ES2 treated with cisplatin, Pt(Ⅳ) and NP@Pt(Ⅳ). The proliferation ability of the ovarian cancer cells were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cellular apoptosis was assessed by flow cytometry. Collection of primary ovarian cancer tissues from patients with primary high-grade serous ovarian cancer who were surgically treated at the Cancer Hospital of the Chinese Academy of Medical Sciences from October to December 2022. The high-grade serous ovarian cancer patient-derived xenograft (PDX) mice were intravenously injected with Cy7.5 labeled NP@Pt(Ⅳ) followed by in vivo imaging system. Mice were treated with PBS, cisplatin and NP@Pt(Ⅳ). Tumor volume and weight were measured in each group. Necrosis, apoptosis and cell proliferation of tumor tissues were detected by hematoxylin-eosin (HE) staining, TUNEL fluorescence staining and Ki-67 immunohistochemistry staining. Body weight and HE staining of heart, liver, spleen, lung and kidney of mice in each group were measured. Results: The platinum release of NP@Pt(Ⅳ) after 48 hours in reducing environment was 76.29%, which was significantly higher than that of 26.82% in non-reducing environment (P<0.001). The platinum content in ES2 cells after 4 hours and 7 hours of treatment with NP@Pt(Ⅳ) (308.59, 553.15 ng/million cells) were significantly higher than those of Pt(Ⅳ) (100.21, 180.31 ng/million cells) and cisplatin (43.36, 50.36 ng/million cells, P＜0.05). The half inhibitory concentrations of NP@Pt(Ⅳ) in ovarian cancer cells ES2, A2780, A2780DDP were 1.39, 1.42 and 4.62 µmol/L, respectively, which were lower than those of Pt(IV) (2.89, 7.27, and 16.74 µmol/L) and cisplatin (5.21, 11.85, and 71.98 µmol/L). The apoptosis rate of ES2 cells treated with NP@Pt(Ⅳ) was (33.91±3.80)%, which was significantly higher than that of Pt(Ⅳ) [(16.28±2.41)%] and cisplatin [(15.01±1.17)%, P＜0.05]. In high-grade serous ovarian cancer PDX model, targeted accumulation of Cy7.5 labeled NP@Pt(Ⅳ) at tumor tissue could be observed. After the treatment, the tumor volume of mice in NP@Pt(IV) group was (130±98) mm3, which was significantly lower than those in control group [(1 349±161) mm3, P<0.001] and cisplatin group [(715±293) mm3, P=0.026]. The tumor weight of mice in NP@Pt(IV) group was (0.17±0.09)g, which was significantly lower than those in control group [(1.55±0.11)g, P<0.001] and cisplatin group [(0.82±0.38)g, P=0.029]. The areas of tumor necrosis and apoptosis in mice treated with NP@Pt(Ⅳ) were higher than those in mice treated with cisplatin. Immunohistochemical staining revealed that there were low expressions of Ki-67 at tumor tissues of mice treated with NP@Pt(Ⅳ) compared with cisplatin. The change in body weight of mice in NP@Pt(Ⅳ) group was not significantly different from that of the control group [(18.56±2.04)g vs.(20.87±0.79)g, P=0.063]. Moreover, the major organs of the heart, liver, spleen, lung, and kidney were also normal by HE staining. Conclusion: Redox-responsive NP@Pt(Ⅳ), produced in this study can enhance the accumulation of cisplatin in ovarian cancer cells and improve the efficacy of ovarian cancer chemotherapy.

[Performance of vaginal self-sampling high-risk HPV genotyping as primary and combining cytology or viral load as secondary in cervical cancer screening].

Objective: To evaluate the efficacy of high-risk HPV (HR-HPV) genotyping with vaginal self-sampling in primary screening and combining cytology or viral load for HR-HPV positive as secondary screening strategies. Methods: The data referring to HR-HPV genotyping of self-collected sample with mass array matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), HR-HPV viral load of physician-collected sample with hybrid capture Ⅱ (HC-Ⅱ), liquid-based cytology and histology of 8 556 women were from Shenzhen cervical cancer screening trial Ⅱ (SHENCCAST-Ⅱ) conducted between April 2009 and April 2010. The data were reanalyzed to determine the sensitivity and specificity to cervical intraepithelial neoplasia (CIN) of grade 2 or worse (CIN Ⅱ+), CIN of grade 3 or worse (CIN Ⅲ+) when HR-HPV genotyping combining with colposcopy as primary screening strategy based on varied HR-HPV subtype (strategy 1, including 5 sub-strategies: 1a: HPV 16/18 positive; 1b: HPV 16/18/58 positive; 1c: HPV 16/18/58/31/33 positive; 1d: HPV 16/18/58/31/33/52 positive; 1e: any HR-HPV positive). The data were also compared to determine the efficacy of cytology (strategy 2, including 5 sub-strategies: 2a, 2b, 2c, 2d, 2e) or HR-HPV viral load (strategy 3, including 4 sub-strategies: 3a, 3b, 3c, 3d) of physician-collected sample as a triage with HR-HPV genotyping for self-sampling HR-HPV positives. Results: (1) The HR-HPV positive rate was 13.77% (1 178/8 556) in the self-collected samples of 8 556 pregnant women. Of them,the prevalences of HPV 16/18, HPV 16/18/58, HPV 16/18/58/31/33 and HPV 16/18/58/31/33/52 were 3.16% (270/8 556), 5.14% (440/8 556), 6.66% (570/8 556) and 9.81% (839/8 556), respectively. The HR-HPV viral load ≥10 relative light units/control (RLU/CO) was 8.87%(759/ 8 556), while cytological results ≥atypical squamous cell of undetermined signification (ASCUS) were 12.05% (1 031/8 556). (2) The strategy 1e had the highest sensitivities for CIN Ⅱ+, CIN Ⅲ+ which were 92.70% and 94.33%,respectively,among 14 sub-strategies,while the lowest specificity and positive predictive value (PPV). Meanwhile,the required colposcopy referral rates were much higher than other 13 sub-strategies (13.77%). The other 4 sub-strategies of strategy 1 (1a, 1b, 1c, 1d), strategy 1a had the highest specificities for CIN Ⅱ+ and CIN Ⅲ+ (97.92%, 97.69%, respectively), while 1d had the highest sensitivities for CIN Ⅱ+ and CIN Ⅲ+ (88.41%, 92.20%, respectively). (3) Both strategies of referring self-sampling HPV 16/18 positives for immediate colposcopy followed by triage physician-collected sample cytology (≥ASCUS) or viral load (≥10 RLU/CO) for non-HPV 16/18 positives had significantly higher sensitivity and specificity for CIN Ⅱ, CIN Ⅲ+, as well as lower referral rates (strategy 2a and 3a). Additionally, based on these two secondary screening strategies, cumulatively using the other four HR-HPV (HPV 58, 31, 33 and 52) positives as triage for immediate colposcopy showed an enhanced sensitivity. Conclusions: Primary HR-HPV cervical cancer screening strategy based on self-sampling with triage of cytology (≥ASCUS) or viral load (≥10 RUL/CO) provides a good balance among sensitivity, specificity for CIN Ⅱ+ and CIN Ⅲ+ and the number of tests required, referral rates. The efficacy of HR-HPV genotyping combining cytology or viral load secondary screening strategies will have a spiral escalation when HPV 58, 31, 33, 52 are included.

[Evaluation of CIN2+ /CIN3+ risk of different HPV subtypes infection combined with abnormal cytology status].

Objective: To determine the morbidity of cervical intraepithelial neoplasia 2+ (CIN2+ ) and CIN3+ of different human papillomavirus(HPV) subtype infection combined with different cytology status. Methods: The Shenzhen Cervical Cancer Screening Trial Ⅰ & Ⅱ (SHENCCASTⅠ&Ⅱ) are population-based cross-sectional cervical cancer screening studis conducted in Shenzhen and surrounding area from 2008 to 2010. A total of 12 097 women who aged 25-59 years were included in the analysis. All of these women were detected by liquid-based cytology test and several high-risk HPV-DNA tests. The ones with HPV positive or atypical squamous cells of undetermined sign (ASC-US) were sequentially conducted by cervical biopsy vaginoscopy. Finally, 10 805 samples with complete data of hybrid capture 2(HC2), the polymerase chain reaction-based matrix-assisted laser desorption/ionization time-of-flight assay (MALDI-TOF), HPV genotyping detection, cytology and pathology results were analyzed. Results: The top 6 infection rates of HR-HPV in CIN2+ and CIN3+ were HPV16, HPV52, HPV58, HPV33, HPV31, HPV18. The highest constituent ratio of cytology in CIN2+ and CIN3+ was high grade squamous intraepithelial lesion(HSIL). The morbidities of CIN2+ of patients infected with HPV16, HPV31, HPV58, HPV33, HPV18, HPV52 were 41.3%, 31.5%, 30.6%, 28.7%, 28.2%, 17.7%, respectively, while the morbidities of CIN3+ of those were 33.5%, 20.5%, 19.4%, 15.7%, 19.2%, 8.3%, respectively.The morbidities of CIN2+ in negative intraepithelial lesion or malignancy (NILM), ASC-US, low grade squamous intraepithelial lesion (LSIL), atypical squamous cell cannot exclude high-grade squamous intraepithelial lesion (ASC-H), high grade squamous intraepithelial lesion (HSIL), atypical glandular cell (AGC) samples were 0.4%, 6.9%, 11.1%, 36.4%, 82.0%, 16.7%, respectively, while the morbidities of CIN3+ of those were 0.2%, 3.1%, 4.2%, 22.7%, 64.8%, 0.0%, respectively. The morbidities of CIN2+ in NILM combined with HPV16, HPV18, HPV31, HPV33 infection were 12.6%, 13.3%, 15.8% and 11.5%, respectively, while the morbidities of CIN3+ of those were 10.3%, 11.1%, 7.9% and 7.7%, respectively.The morbidities of CIN2+ and CIN3+ in ASC-US combining with hrHPV infection were high, and the top 6 subtypes associated with high risk of CIN2+ were HPV31 (35.7%), HPV33 (26.9%), HPV16 (26.5%), HPV58 (22.4%), HPV52 (18.6%), HPV68 (15.4%), while those associated with high risk of CIN3+ were HPV16 (20.4%), HPV31 (14.3%), HPV33 (11.5%), HPV58 (8.6%), HPV68 (7.7%), HPV52 (5.8%). Conclusions: Cytology combined with HPV genotyping detection can more effectively estimate the morbidity risks of CIN2+ and CIN3+ . Both high prevalence rates and high risks associated with CIN2+ and CIN3+ of HPV31, HPV33, HPV52 and HPV58 are observed. NILM and ASC-US status combined with these subtypes mentioned above are advised to be conducted by colposcopy.

Effect of matrine on reducing damage to bovine mammary epithelial cells induced by Staphylococcus aureus alpha-hemolysin.

Taking bacterial virulence factors as targets is a new therapy for treating host bacterial infection. The aim of this study was to investigate the effect of matrine on α-hemolysin production of Staphylococcus aureus (S. aureus) and reducing the damage to bovine mammary epithelial cells (BMECs) induced by S. aureus α-hemolysin. Subinhibitory concentrations of matrine decreased the production of α-hemolysin in none dose-dependent manner and matrine exhibited a protective effect on S. aureus-induced BMECs injury. The results indicated that the structure of matrine may potentially be used as a basic structure for development of drugs aimed at curing and preventing dairy bovine mastitis.

[Application of da Vinci robotic surgical system in radical resection of gastric and colorectal cancer: a report of 647 cases].

OBJECTIVE: To investigate the feasibility and safety of robotic-assisted radical resection of gastric and colorectal cancer. METHODS: The clinical data of 305 patients who received radical resection of gastric cancer and 342 patients who received radical resection of colorectal cancer both accomplished by the da Vinci robotic surgical system at the Southwest Hospital, Third Military Medical University from March 2010 to December 2014 were retrospectively analyzed. In gastric cancer group, radical total gastrectomy were performed in 69 cases, proximal gastrectomy in 11 cases, distal gastrectomy in 213 cases and 12 cases of gastric stump cancer.In colorectal cancer group, radical resection of the right colon were performed in 6 cases, left colon in 4 cases, transverse colon in 1 case, sigmoid colon low anterior resection procedure in 24 cases, rectal low anterior resection procedure in 222 cases, abdominoperineal excision procedure in 79 cases and Hartmann procedure in 6 cases. RESULTS: All cases had robotic-assisted radical resection successfully. In gastric cancer group, the mean operation time was (226±62) minutes, the mean blood loss was (125±77) ml, the mean number of harvested lymph nodes was 34±10; the mean time for patients taking normal activity was (3.2±1.5) days, the mean time for gastrointestinal function recovery was (3.1±1.3) days, the mean time for taking liquid food was (3.5±1.9) days. The mean hospitalization was (7.9±3.7) days postoperatively. In colorectal group, the mean operation time was (181±61) minutes, the mean blood loss was (110±93) ml, the mean number of harvested lymph nodes was 19±6; the mean time for patients taking normal activity was (2.9±1.5) days, passage of first flatus was (2.7±1.7) days. The mean hospitalization was (7.1±1.6) days postoperatively. Surgical complications occurred in 28 patients (9.2%) of gastric cancer group and 30 patients (8.8%) of colorectal cancer group, all the cases were recovery before leaving hospital with non-operation therapy. The short-term efficiency was obvious. CONCLUSION: Robotic-assisted radical resection for gastric and colorectal cancer is a feasible and safe surgical procedure combined the minimal trauma and fast recovery.

The tissue selecting technique (TST) versus the Milligan-Morgan hemorrhoidectomy for prolapsing hemorrhoids: a retrospective case-control study.

BACKGROUND: Milligan-Morgan hemorrhoidectomy (MMH) is the procedure of choice in the management of hemorrhoidal disease. However, this procedure is associated with significant postoperative pain. Tissue selecting technique (TST) is a segmental stapled hemorrhoidopexy, which aims to reduce the postoperative pain, rectovaginal fistula (RVF) and rectal stenosis. The aim of the present study was to compare the clinical outcomes between TST and MMH. METHODS: A case-control study was undertaken to investigate the difference in clinical characteristics between the patients treated with TST and those treated with MMH. Intraoperative and postoperative parameters in both groups were collected and compared. RESULTS: One hundred and ninety-five eligible patients underwent either TST (n = 121) or MMH (n = 74). The pain score was significantly less in the TST group than that in the MMH group at the first defecation and at 12 h, day 3 and day 7 postoperatively (P = 0.001). Further analysis revealed that, at the time point of 12 h, day 3, day 7 and during first defecation, the pain score in the TST group and TST + STE group was less than that in the MMH group (P = 0.001). No patient in either group developed postoperative rectal stenosis. Furthermore, no case of RVF was identified in the TST group. The 1-year recurrence rate was 3.3 % (4/121) and 2.7 % (2/74), respectively, in TST and MMH groups (P = 1.0). CONCLUSIONS: The 1-year recurrence rate after TST and MMH for the treatment of patients with grade III-IV hemorrhoids is similar. It is encouraging that TST is associated with less postoperative pain and no RVF or rectal stenosis.

Botulinum toxin injection versus lateral internal sphincterotomy for chronic anal fissure: a meta-analysis of randomized control trials.

BACKGROUND: Although surgery is the gold standard treatment for anal fissure, the main concern remains its side effects and complications. Botulinum toxin injection and lateral internal sphincterotomy are technical options for patients suffering from chronic anal fissure. However, little is known about the efficacy of these two techniques. The aim of this meta-analysis was to compare the outcomes of botulinum toxin injection versus lateral internal sphincterotomy for chronic anal fissure. METHODS: Original studies in English were searched from the MEDLINE database, PubMed, Web of Science, and the Cochrane Library database. Randomized control trials that compared botulinum toxin injection with lateral internal sphincterotomy were identified. Data were independently extracted for each study, and a meta-analysis was performed using fixed and random effects models. RESULTS: Four hundred and eighty-nine patients from seven trials met the inclusion criteria. Patients undergoing lateral internal sphincterotomy had a higher-healing and incontinence rate. No statistically significant differences were noted in total complications between botulinum toxin injection and lateral internal sphincterotomy. Patients treated with lateral internal sphincterotomy had a significantly lower recurrence rate than the patients treated with botulinum toxin injection. CONCLUSIONS: Our meta-analysis shows that lateral internal sphincterotomy was superior to botulinum toxin injection in terms of healing rate and lower recurrence rate. Botox, however, is safe associated with a lower rate of incontinence and could be used in certain situations. Further studies with a long-term follow-up are required to confirm our observations.

Effects of CO(2) pneumoperitoneum on peritoneal macrophage function and peritoneal metastasis in mice with gastric cancer.

BACKGROUND: Whether laparoscopy with CO(2) pneumoperitoneum affects the peritoneal metastasis of gastric cancer is a pressing question. In light of the important impact change in peritoneal macrophage function has on the peritoneal metastasis of gastric cancer, this study investigated the change in peritoneal macrophage function in gastric cancer in the CO(2) pneumoperitoneum environment, as well as its effect on the peritoneal metastasis of gastric cancer. METHODS: An orthotopic transplantation model of murine forestomach carcinoma was established using the 615 mouse line. The mice bearing tumors were randomly divided into four groups (30 mice each group): anesthesia alone, laparotomy, mini-laparotomy, and CO(2) insufflation. After the operation, peritoneal macrophages were collected from 6 mice in each group and cultured. The phagocytosis of neutral red by macrophages and the levels of NO, TNF-α, IL-10, and VEGF produced by macrophages were measured after 12, 24, 48, and 72 h of culture. The remaining mice were observed after 2 weeks for the rate of peritoneal metastasis of forestomach carcinoma cells and the total weight of implanted nodules. RESULTS: In the laparotomy group, 4 mice died intraoperatively and 2 died in the CO(2) insufflation group. The uptake of neutral red by peritoneal macrophages and the levels of NO, TNF-α, IL-10, and VEGF secreted by peritoneal macrophages in the laparotomy group and mini-laparotomy group after 12 h of culture were all significantly higher than those in the anesthesia-alone group (p < 0.05). The corresponding levels in the CO(2) insufflation group after 12 h were all significantly lower than those in the anesthesia-alone group (p < 0.05). There were no significant differences among the four groups at 24, 48, and 72 h after culture. Comparing with those in the laparotomy group, the uptake of neutral red by peritoneal macrophages and the levels of NO, TNF-α, IL-10, and VEGF secreted by peritoneal macrophages in the CO(2) insufflation group were all significantly lower after 12 h of culture (p < 0.05), but did not differ significantly at 24, 48, and 72 h of culture (p > 0.05), and did not differ significantly in the mini-laparotomy group at all the time (p > 0.05). The rate of peritoneal metastasis of mouse forestomach carcinoma was 50% in the laparotomy group, 45.83% in the mini-laparotomy group, and 45.45% in the CO(2) insufflation group; this difference was not statistically significant (p > 0.05). The total weight of implanted nodules of mouse forestomach carcinoma was 1.02 ± 0.38 g in the laparotomy group, 0.97 ± 0.41 g in the mini-laparotomy group, and 0.93 ± 0.45 g in the CO(2) insufflation group, which was not a statistically significant difference (p > 0.05). CONCLUSION: CO(2) pneumoperitoneum neither significantly changes the phagocytosis and cytokine secretion functions of peritoneal macrophages in gastric cancer-bearing mice nor significantly promotes peritoneal metastasis of gastric cancer.

[A prospective randomized controlled study of the therapeutic effects of a self-developed novel low-temperature boric acid wet dressing on the face and neck wounds of patients with superficial burns].

Objective: To observe and explore the clinical therapeutic effects of a self-developed novel low-temperature boric acid wet dressing on the face and neck wounds of patients with superficial burns. Methods: A prospective randomized controlled study was conducted. One hundred cases of superficial burn (sunburn) patients who met the inclusion criteria were admitted to Department of Dermatology of Chongqing Hospital of Traditional Chinese Medicine from October 2016 to June 2018, the course of sunburn was less than 15 days. According to the of random number table, the patients were divided into new dressing (ND) group (30 males and 20 females, aged (55±14) years) and conventional dressing (CD) group (28 males and 22 females, aged (59±12) years). Patients in ND group were treated with a self-developed new low-temperature boric acid wet dressing, and patients in CD group were treated with normal temperature boric acid solution wet dressing, 3 times a day. The Eczema area and severity index (EASI) score and Visual Analogue Scale (VAS) score of patients before and 14 days after treatment (patients discharged from hospital within 14 days were recorded on the day of discharge), the number of cured patients at 6, 8, 10, and 11 to 14 days after treatment, and the efficacy were compared between the patients in 2 groups. The nurse operators were investigated by self-made convenience questionnaire, and the time and convenience required of 2 dressings were compared. Data were statistically analyzed with independent sample t test or Mann-Whitey U test. Results: Before the treatment, the EASI and VAS score in those two groups showed no significantly difference (t=1.576, 1.492, P>0.05). At 14 days after treatment, the EASI score (2.4±0.4) points in ND Group was significantly lower than (4.6±0.7) points in CD Group (t=3.552, P<0.01); the VAS score (0.51±0.12) points in ND Group was significantly lower than (0.98±0.19) points in CD Group (t=3.496, P<0.01). At 14 days after treatment, the cured time of sunburns in ND Group was significantly shorter than that in CD Group (Z=-6.690, P<0.01); the treatment effects of ND Group showed better than that in CD Group (Z=3.387, P<0.01). The time for nurses operating ND was significantly shorter than that in CD (Z=-5.575, P<0.01); the nurses also believed the operation of ND was more convenient than CD (Z=-4.304, P<0.01). Conclusions: Compared with that of CD, the application of ND can shorten the recovery time and improve the treatment efficiency. At the same time, the application of ND can significantly reduce the time of nursing operations, and the material is easy to use. This new material is worthy of clinical promotion for the treatment of superficial burns.

[Study on the damage of the tight junctions of nasal mucosal epithelial cells by artemisia annua pollen].

Objective: To investigate the damage and mechanism of artemisia annua pollen on tight junction of human nasal mucosa epithelial cells (HNEpC). Methods: HNEpC were cultured in vitro. Different concentrations of artemisia annua pollen (0, 20, 40, 80, 100, 160, 200 µg/ml) were used to intervene the cells for 24 h, and the cell proliferation activity was detected by the CCK-8 method. The expression and phosphorylation of p38MAPK signaling pathway were detected by Western Blot before and after the intervention of SB203580, a p38MAPK inhibitor in HNEpC. Immunofluorescence chemical staining, Western Blot and quantitative real-time PCR (qPCR) were used to observe the expression and distribution of tight junctions Occludin and Claudin-1. SPSS 21.1 software was used for statistical analysis. Results: CCK-8 results showed that, compared with the control group, the proliferation activity of HNEpC increased after 6 h intervention with different concentrations of artemisia annua pollen (all P<0.05). After 12 h of intervention, the proliferation activity of HNEpC in the 20, 40, 80, 100 and 160 µg/ml groups was not significantly changed (all P>0.05), while that in the 200 µg/ml group was decreased (P<0.05). After the intervention for 24 h, the proliferation activity of cells in the 20 and 40 µg/ml groups was not significantly changed (all P>0.05), while that in the 80, 100, 160 and 200 µg/ml groups was decreased (all P<0.05). Immunofluorescence staining showed that the Occludin and Claudin-1 proteins in the normal control group were localized on the cell membrane and expressed more and formed a ring structure around the cell membrane. However, under the intervention of high concentration artemisia annua pollen, its expression level decreased, appeared broken, fuzzy, and nonuniform distribution. Western Blot and qPCR results showed that after 24 h of intervention, the expression levels of HNEpC Claudin-1 protein and its mRNA in the pollen groups (40, 80, 100, 160, 200 µg/ml) of artemisia annua decreased compared with those of those of the control group (mRNA expression levels were 0.567±0.214, 0.443±0.109, 0.462±0.160, 0.497±0.134, 0.388±0.076 compared with 1.001±0.067, respectively, all P<0.05). However, the mRNA of Occludin protein and its mRNA only decreased in the 200 µg/ml treatment group (mRNA expression level was 0.631±0.109 compared with 1.016±0.026, P<0.05), while all the other treatment groups increased (mRNA expression levels were 1.258±0.134, 1.827±0.103, 2.429±0.077, 1.707±0.085, 1.477±0.066 compared with 1.016±0.026, respectively, all P<0.05). Western Blot showed that p-p38MAPK expression increased after intervention with 100, 160, 200 µg/ml artemisia annua pollen for 24 h. SB203580 could inhibit the decreasing expression of Occludin caused by artemisinin pollen (mRNA expression was 1.255±0.179 compared with 0.631±0.109, P<0.05), but had no effect on Claudin-1 protein expression. Conclusion: Pollen from artemisia annua may activate p38MAPK signaling pathway and destroy the close connection of HNEpC.

[Analysis on the technical characteristics and clinical efficacy of robotic-assisted intersphincteric resection for patients with low rectal cancer].

Objective: To explore the technical characteristics and short-term clinical efficacy of robotic-assisted intersphincteric resection (ISR) for patients with low rectal cancer. Methods: A retrospective cohort study was used. Inclusion criteria: (1) rigid colonoscopy showed lower margin of the tumor ≤5 cm from the anal verge; (2) preoperative rectal MRI or endorectal ultrasound revealed staging T1-2, or T3 patients receiving concurrent chemoradiotherapy; (3) patients less than 70 years old with good function of anal sphincter before surgery; (4) no synchronous multiple primary carcinoma, and no distant metastasis; (5) the method of operation was agreed by the patient. Exclusion criteria: (1) T4 stage tumors; (2) sphincter dysfunction before operation; (3) recurrent tumors; (4) lower edge of tumors beyond the dentate line; (5) death due to non-rectal cancer during follow-up and unsatisfactory follow-up data. The clinical data of 21 patients with low rectal cancer meeting inclusion criteria undergoing robotic-assisted ISR at our department from January 2015 to June 2018 were collected. Parameters during and after operation were observed. Anorectal manometry was performed at 3, 6, and 12 months after the operation, and anal function was evaluated at 3, 6, and 12 months after the closure of the stoma by Kirwan classification and Wexner fecal incontinence score. The key steps of the operation are as follows: according to the principle of total mesorectal excision, the robot continued to enter into the levator ani hiatusdistally, and dissectin the sphincter space; according to the scope of sphincter resection, ISRwas divided into partial ISR, subtotal ISR, and total ISR; subtotal and total ISR usually needed to be combined with transanal pathway. The reconstruction of digestive tract was performed by double stapler anastomosis under laparoscope orhand-sewnanastomosis under direct vision, and preventive ileostomy was completed in the right lower abdomen. Results: Of 21 patients, 13 were male and 8 were female with mean age of (57.5±16.3) years. All the patients successfully completed the operation without conversion to laparotomy. Fourteen cases (66.7%) adopted partial ISR through complete transabdominal approach, 6 cases (28.6%) adopted the subtotal ISR through combined transabdominal and transanal approachs, and 1 case (4.8%) adopted the total ISR through the combined transabdominal and transanal approachs. The total operation time was (213.1±56.3) minutes, including (27.3±5.4) minutes for mechanical arm installation and (175.7±51.6) minutes for robotic operation. The amount of intraoperative hemorrhage was (62.8±23.2) ml, and no blood transfusion was performed in any patient. All patients underwent prophylactic ileostomy, and the stoma was closed 3-6 months after the operation. Except one case of anastomotic leakage, all other stomas were closed successfully. The postoperative hospitalization time was (7.6±2.2) days, and time to fluid intake was (3.3±0.9) days. One case of anastomotic leakage, one case of anastomotic stenosis, one case of inflammatory external hemorrhoids and one case of urinary retention occurred after surgery,and all of them were cured by conservative treatment. The mean diameter of tumors was (2.9±1.2) cm, and the number of harvested lymph node was 12.8 ± 3.3. In the whole group, the circumcision margin was negative, the proximal margin was (12.2 ± 2.1) cm, the distal margin was (1.1 ± 0.4) with all negative, and the R0 resection rate was 100%. The results of anorectal manometry showed that the preoperative rest pressure, rectal maximum squeeze pressure, initial sensory volume and maximum tolerated volume were (45.19±8.46) mmHg, (128.18±18.80) mmHg, (44.33±10.11) ml and (119.00±19.28) ml, respectively;these parameters reduced significantly 3 months after operation and they were (23.44±5.54) mmHg, (93.72±12.15) mmHg, (17.72±5.32) ml and (70.44±10.9) ml, respectively. The differences were statistically significant (all P<0.001). The resting pressure and the rectal maximum squeeze pressure returned to preoperative levels 12 months after operation, which were (39.33±6.64) mmHg and (120.58±16.47) mmHg, respectively (both P>0.05), while the initial sensory volume and the maximum tolerated volume failed to reach the preoperative state, which were (30.67±7.45) ml and (92.25±10.32) ml, respectively (both P<0.05). The patients were followed up for (22.1±10.6) months without local recurrence and distant metastasis. Eighteen patients were evaluated for anal function: Kirwan classification was grade I for 6 cases, grade II for 7 cases, grade III for 4 cases, and grade IV for 1 case; Wexner incontinence score was 8.6±0.8; 14 cases had good defecation control. Conclusion: The clinical efficacy of ISR with Da Vinci robot in the treatment of low rectal cancer is satisfactory.

Evaluation of the COBAS® TaqMan® system in patients with low hepatitis B virus DNA undetectable with PCR assay kit.

OBJECTIVE: We conducted a comparison of the diagnostic kit for quantification of hepatitis B virus DNA (PCR-fluorescence probing) and COBAS TaqMan automated nucleic acid extraction and real-time polymerase chain reaction (PCR) amplification systems. We tested their capacity to quantify and diagnose patients with chronic viral hepatitis B with low viral load < 1 x 103 IU/mL, in hope to provide further evidence for promoting the application of COBAS TaqMan as the diagnostic method for such patients. PATIENTS AND METHODS: Diagnostic kit and COBAS TaqMan system were tested on 100 patients diagnosed with chronic viral hepatitis B in our hospital and with a viral load lower than the detection limit of real-time extraction-quantification kit. These patients included 47 cases with chronic viral HBV, 53 cases of HBV-associated cirrhosis (11 cases were HBV-associated liver cancer with cirrhosis). COBAS TaqMan real-time quantification PCR with a sensitivity of 20 IU/ml was performed to test the reproducibility for the diagnosis result. RESULTS: The COBAS TaqMan real-time system quantified 76 cases out of 100 with a viral load higher than 20 IU/ml (detection rate, 76%). Among these patients, there were 33 cases of chronic viral HBV (without cirrhosis) (detection rate, 70.2%), 43 cases of cirrhosis (detection rate, 81.1%, including 28 cases of compensatory cirrhosis and 15 cases of decompensated cirrhosis), and 11 cases of liver cancer (detection rate, 81.2%). CONCLUSIONS: The COBAS TaqMan system has higher sensitivity than traditional real-time PCR detection kit, especially for HBV-related cirrhosis and liver cancer with low viral load. The limitation of real-time PCR should be taken into account during treatment monitoring and the alternative of COBAS TaqMan system should be promoted in patients with high risk of liver cirrhosis and cancer to avoid delayed diagnosis and improve clinical outcome.

Electrochemical study of the effect of ADP and AMP on the kinetics of glutamate dehydrogenase.

A chronoamperometric method based on the 'diffusion' layer concept of the convective system was used to assay the glutamate dehydrogenase (GLDH) activity. Once the reaction was initiated by adding the enzyme GLDH into a well-stirred nicotinamide adenine dinucleotide (NADH, coenzyme) solution, the steady-state oxidation limiting current of NADH would decrease linearly in a short time. The major advantage of this method is that it directly indicates the continuous in-situ change of the coenzyme concentration, thus, the real initial reaction rate of enzyme-catalyzed reaction, V0, can be determined. Using this method, the effect of adenosine-5'-monophosphate (AMP) and adenosine-5'-diphosphate (ADP) on the GLDH activity has been monitored. The results showed that ADP and AMP could increase the activity of GLDH. This activation mechanism was proposed by the voltammetric study.