Objective To investigate the expression of cytokeratin 34βE12 in esophageal squamous cell carcinoma (ESCC),and its mechanism of action in the process of occurrence and development of an ESCC.Methods Immunohistochemistry was used to analyze the expression of cytokeratin 34βE12 in 252 ESCC patients,66 patients with esophageal carcinoma in situ,and 106 patients with adjacent normal esophageal mucosa before the relationship between its expression and biological behavior was evaluated on the basis of complete clinical information.In addition,Western blotting was used to determine the expression of cytokeratin 34βE12 in 60 patients with esophageal cancer and adjacent normal esophageal tissues.Results (1)The positive rate of caveolin-1 in ESCC,carcinoma in situ,and adjacent normal tissues was 85.7％,54.5％,and 25.7％,respectively.The difference between them was statistically significant (P ＜0.01).(2)The positive rate of cytokeratin 34βE12 in stages Ⅰ,Ⅱ,Ⅲ of ESCC was 76.5％,84.7％,and 96.3％,respectively.The expression intensity of cytokeratin 34βE12 in carcinoma tissue was gradually increased with the advance of clinical stages with a statistically significant difference (P =0.038).The positive rate of cytokeratin 34βE12 with group of lymph node metastasis was significantly higher than those without lymph node metastasis (P ＜ 0.01).(3)Western blotting results further confirmed that the expression of cytokeratin 34βE12 in ESCC was significantly higher than that in adjacent normal esophageal tissue (P ＜0.01).Conclusions The high expression of caveolin-1 might be involved in the occurrence and development of esophageal cancer.The expression of cytokeratin 34βE12 was correlated with the clinical stage of esophageal cancer.cytokeratin 34βE12 was a potential therapeutic target and a valuable prognostic indicator of esophageal cancer progression.

@@

OBJECTIVETo establish a method for quick finding of the absorption ingredients of Paeoniae Radix Alba in order to select the index of quality control.

METHODThe absorption ingredients of three concentration of Paeoniae Radix Alba were investigated with the in vitro-everted intestinal sac (VEIS) model. The intestinal sac fluids of jejunum and ileum were collected in different time and detected by HPLC. The accumulative absorption quantity of albiflorin and paeoniflorin were calculated, respectively.

RESULTFive ingredients could be detected. In different concentrations of Paeoniae Radix Alba, albiflorin and paeoniflorin in various intestinal sections were the linear absorption (R2 > 0.9), conformed to the zero order absorption rate. The values of Ka in the jejunum and ileum were increased along with the raised dosage of the Paeoniae Radix Alba (P < 0.05), indicating a passive absorption manner.

CONCLUSIONSEMAC could be used as a tool to find the absorption ingredients of Paeoniae Radix Alba. Compared with the jejunum, the ileum could provide the more absorption information. It was showed that the optimal detecting time was 60 min.

Animals ; Intestinal Absorption ; Male ; Paeonia ; Rats ; Rats, Wistar

OBJECTIVETo establish a method for quick investigating the absorption ingredients of Plantaginis semen and guiding the index selection for its quality control.

METHODThe absorption of three concentrations of Plantaginis semem was investigated with the in vitro everted intestinal sac (VEIS) model The intestinal sac contents of jejunum and ileum were collected at different time and geniposidic acid was detected by HPLC and LC-MS(n) as the representative marker.

RESULTSix ingredients could be detected. At different concentrations of Plantaginis semen, geniposidic acid tested by VEIS showed that there was a good linear correlation between the drug absorption from the medium across the intestinal epithelium into the sac contents in various intestines section. The absorption of the gut sacs from 0 to 90 min manifested a significant time-dependent manner. The Ka of geniposidic acid in the jejunum and ileum increased along with the raised dosage of the Plantaginis Semen (P < 0.05), which indicated a passive absorption manner.

CONCLUSIONThis method can be used as a tool to investigate the absorption ingredients of Plantaginis Semen. Comparing with the jejunum, the ileum can provide more absorption information faster. The optimal incubation time in intestinal sac was 90 min.

Animals ; Body Fluids ; chemistry ; metabolism ; Drugs, Chinese Herbal ; pharmacokinetics ; Ileum ; chemistry ; drug effects ; metabolism ; Intestinal Absorption ; Jejunum ; chemistry ; drug effects ; metabolism ; Male ; Models, Biological ; Plantago ; chemistry ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the distribution features of anti-neutrophil cytoplasmic antibody (ANCA) in patients with cardiovascular and cerebrovascular diseases and the clinical characteristics of the patients.Methods:Clinical data of 6 759 patients who were treated in Baoding No.1 Central Hospital for cardiovascular and cerebrovascular diseases during January 2015 to July 2019 were collected and analyzed. ANCA was detected by indirect immunofluorescence (IIF). Antibodies against myeloperoxidase (MPO) and protease 3 (PR3) were detected by enzyme-linked immunosorbent assay (ELISA).Results:IIF showed that 558 out of the 6 759 cases tested positive for ANCA with a positive rate of 8.26%. Among them, 382 (68.46%) were positive for perinuclear ANCA (p-ANCA) and 176 (31.54%) were positive for cytoplasmic ANCA (c-ANCA). Anti-MPO and anti-PR3 antibodies were detected in 69 ANCA-positive cases, while the antibodies against other target antigens were detected in 489 cases. The ratio between the two groups was 1∶7.09. The incidence of recurrent respiratory tract infection and pulmonary interstitial lesions in ANCA-positive cases was 69.35% (387/558) and 64.52% (360/558), respectively, which was significantly higher than that in ANCA-negative patients [40.51% (2 512/6 201) and 33.17% (2 057/6 201)].Conclusions:Anti-MPO and anti-PR3 antibody detection could not replace IIF to detect total ANCA for a high rate of missed diagnosis would be caused. Early detection of ANCA would be of great significance to patients with chronic cardiovascular and cerebrovascular diseases.

Objective To evaluate the effect of ulinastatin on postoperative cognitive dysfunction in preoperative sleep deprived aged rats.Methods Sixty clean healthy male Sprague-Dawley rats,aged 18 months,weighing 350-500 g,were divided into 3 groups (n=20 each) using a random number table:control group (group C),sleep deprivation group (group D) and ulinastatin group (group U).Sleep deprivation was induced by using modified multiple platform method in D and U groups,and then splenectomy was performed in three groups.Ulinastatin 100 U/g was intraperitoneally injected before sleep deprivation and immediately after operation in group U.Ten rats were randomly selected at 3 days after operation and sacrificed,and hippocampi were removed for determination of the contents of interleukin-1β (IL-1β),IL-6 and tumor necrosis factor-α (TNF-α) by enzyme-linked immunosorbent assay.Morris water maze test was performed at 3-7 days after operation in the rest ten rats in each group,and the escape latency and time of staying at the original platform were recorded.Results Compared with group C,the escape latency was significantly prolonged at 4-6 days after operation,the time of staying at the original platform was shortened,and the contents of IL-1β,IL-6 and TNF-α were increased at 3 days after operation in D and U groups (P＜0.05).Compared with group D,the escape latency was significantly shortened at 4-6 days after operation,the time spent in the original platform was prolonged,and the contents of IL-1β,IL-6 and TNF-α were decreased at 3 days after operation in group U (P＜0.05).Conclusion Ulinastatin can mitigate postoperative cognitive dysfunction in preoperative sleep deprived aged rats,which is related to inhibiting inflammatory responses.

Objective:To analyze the relationship between perineural invasion and other clinicopathological factors and its effect on the prognosis of gastric cancer.Methods:The clinicopathological data of 665 patients with gastric cancer were retrospectively analyzed. According to the presence of perineural invasion, the patients were divided into perineural invasion positive group and perineural invasion negative group. The relationship between perineural invasion and other clinicopathological factors and its effect on the prognosis of gastric cancer were analyzed. After eliminating the potential confusion bias between the two groups by propensity score matching (PSM) , the differences of 5-year cumulative survival rate between the two groups of gastric cancer patients were compared.Results:The incidence of perineural invasion was 17.0% (113 cases) . The binary logistic regression analysis showed that the depth of tumor invasion and vascular tumor thrombus were independent factors influencing the occurrence of gastric cancer perineural invasion (all P<0.001) . Univariate analysis showed that age (>60 years) , tumor diameter (>4 cm) , borrmann classification, depth of invasion, lymph node metastasis, TNM stage, degree of differentiation, vascular tumor thrombus, perineural invasion, tumor nodule, tumor site, resection site, and surgical operation were the influencing factors for the prognosis of patients with gastric cancer ( P<0.05) , but multivariate analysis showed that age (>60 years) , tumor diameter (>4cm) , depth of invasion, lymph node metastasis, and positive vascular tumor thrombi were independent risk factors affecting the prognosis of gastric cancer patients ( P<0.05) .However, perineural invasion cannot be an independent factor influencing the poor prognosis of gastric cancer in a multivariate analysis. Survival analysis was performed after propensity matching scores, and it was found that there was no statistically significant difference in the five-year survival rate between the perineural invasion positive group and the perineural invasion negative group (34.6% vs 43.0%; χ2=1.713; P=0.191) ,and there was no significant difference in the survival curve analysis between the two. Conclusion:Most patients with gastric cancer of perineural invasion have poor prognosis, but perineural invasion cannot be an independent prognostic factor for the prognosis of gastric cancer.

Radiation-induced brain injury (RBI) is the most serious complication of head and neck tumor after radiotherapy. The pathogenesis of RBI is complicated, and the clinical course is irreversible, while no effective treatment available. The activation of glial cells is one of the main theories of RBI, and the prevention and treatment of RBI by targeting glial cells is the focus of current research. As a post-transcriptional regulatory factor, microRNA (miRNA) has been confirmed to be involved in regulatingglial cell radiosensitivity, inflammation type transformation, autophagy, exosomatic, long non-coding RNA (lncRNA), circular RNA (circRNA) and other related pathways, thereby mediating the occurrence and development of cascade reaction of inflammatory injury and neurological function repair of central nervous system (CNS) disease. Therefore, the establishment of miRNA - glial regulatory network may provide a new strategy for the prevention and treatment of RBI.

Objective:To study the effect of enhanced recovery after surgery (ERAS) on intestinal function and gut microbiota changes in patients who underwent laparoscopic gastrectomy.Methods:From Aug. 2018 to Dec. 2019, 80 patients who underwent laparoscopic radical D2 gastrectomy for gastric cancer in the first Department of Gastrointestinal Surgery of Yantai Yuhuangding Hospital were selected. According to whether it adopts ERAS treatment or not, patients were divided into 2 groups (n=40) : ERAS group and traditional perioperative treatment group. The time of postoperative bowel sounds, the time of first exhaust and defecation, the proportion of antibiotic-related diarrhea and surgical site infection (SSI) were recorded. Stools were collected before operation, first time after operation, 1, 2 weeks and 1 month after operation. 16S rRNA sequencing method was used to identify the diversity and species of gut microbiota. The diversity index of intestinal flora in the perioperative period and changes in the proportion of probiotics (bifidobacterium and lactobacillus) were compared.Results:The appearance time of bowel sounds, the first exhaust and defecation time [ (16.25±6.41) h, (23.95±6.02) h, (34.95±9.34) h] in ERAS group were significantly earlier than those in the traditional treatment group [ (22.3±6.49) h, (28.45±7.12) h, (48.1±15.64) h], and the difference was statistically significant ( P<0.05) . The incidence of antibiotic-related diarrhea was higher in the traditional treatment group (3/40) than in ERAS group (1/40) , but the difference was not statistically significant ( P>0.05) . The ratio of postoperative SSI was slightly higher in ERAS group, but the difference was not statistically significant ( P>0.05) . In the perioperative period, the intestinal flora diversity index (Chao1 and Shannon index) and the proportion of probiotics (lactobacillus acidophilus and bifidobacterium) were not significantly different between the two groups before surgery ( P>0.05) ; while at the first time, one week, 2 weeks after the operation, and 1 month after the operation, ERAS group was higher than the traditional group ( P<0.05) ; and at each postoperative time point, the traditional group decreased significantly than the ERAS group. The first time decrease was the largest, ( P<0.05) ; With the passage of time after operation, the diversity of intestinal flora and the proportion of probiotics gradually recovered. By 1 month after operation, the two groups did not return to the preoperative gut microbiota diversity state or proportion. Conclusion:The concept of enhanced recovery after surgery (ERAS) promotes the recovery of intestinal function in patients with gastric cancer, does not reduce the proportion of antibiotic-associated diarrhea (AAD) or surgical site infections (SSI) , and maintains the diversity of gut microbiota balance and stability.

【Objective】 To explore the role and mechanism of TRPC in promoting extracellular matrix (ECM) deposition in rat glomerular mesangial cells (HBZY-1). Methods Immunofluorescence staining was performed to observe the distribution and expression of TRPC1 and TRPC6 in HBZY-1 cells. After AngⅡ stimulation, qRT-PCR and Western blotting were used to detect the mRNA and protein expressions of Gαq/PLCβ4/TRPC signaling pathway main proteins and ECM deposition indicators (α-SMA, collagenⅢ and fibronectin). By silencing the expressions of TRPC1 and TRPC6 by RNA interference, the expressions of ECM deposition indicators were detected. Changes in [Ca²⁺]i influx were determined through Fluo-4AM Ca²⁺ imaging. 【Results】 Both TRPC1 and TRPC6 were expressed in HBZY-1, and were mainly located in cell membrane and cytoplasm. After AngⅡ stimulation, Gαq/PLCβ4/TRPC signaling pathway was activated, and the mRNA and protein expressions of Gαq, PLCβ4, TRPC1 and TRPC6 were all increased (P<0.05). [Ca²⁺]i influx also increased (P<0.01), and the mRNA and protein expressions of ECM deposition indicators (α-SMA, ColⅢ and Fn) were upregulated (P<0.05). Silencing the expressions of TRPC1 and TRPC6 by RNA interference led to decreased [Ca²⁺]i influx (P<0.05), and downregulated mRNA and protein expressions of ECM deposition indicators in HBZY-1 cells (P<0.05). The results suggested that inhibition of TRPC expressions could inhibit AngⅡ induced ECM deposition in HBZY-1 cells, which might be associated with decreased [Ca²⁺]i influx. 【Conclusion】 TRPC may be a novel therapeutic target of renal fibrosis.