Application of reversely connecting circular stapler technique in cervical esophagogastric anastomosis.

Introduction: The incidence of anastomotic leakage is relatively high (6-26%) in esophagogastrocervical anastomosis. The traditional anastomotic mode has high tissue tension in the process of anastomosis, which can easily cause tissue tear in the anastomotic area and increase the incidence of anastomotic leakage. Aim: To investigate the usefulness of reversely connecting circular stapler technique for reducing anastomotic tension and injury of the esophageal and gastric wall, preventing anastomotic leakage in minimally invasive McKeown esophagectomy. Material and methods: A total of 133 patients with esophageal cancer who underwent minimally invasive McKeown esophagectomy from January 2021 to December 2021 were collected. Characteristics, surgical data, and outcomes of patients were recorded and retrospectively analyzed. There were 83 patients in the reverse order connecting circular stapler group and 50 patients in the conventional order connecting group. Results: Anastomotic leakage was observed in 2 (2.4%) cases in the reverse order connecting circular stapler group. Compared with the conventional connection group, the reverse connecting group had lower incidence of anastomotic leakage, shorter anastomotic time, shorter hospital stay, and lower incidence of pulmonary and chest infections and hoarseness. Conclusions: The reversely connecting circular stapler technique could reduce the incidence of anastomotic leakage. The technique is simple to perform and easy to learn. Therefore, it is useful for the prevention of anastomotic leakage.

AHSG, a Gene Promoting Tumour Proliferation, Migration and Invasion, is an Independent Prognostic Factor for Poor Overall Survival in Lung Adenocarcinoma.

OBJECTIVE: Recent studies have shown that the metabolic process-related gene AHSG is involved in multiple pathological processes of tumours. This study will explore the relationship between AHSG and lung adenocarcinoma. MATERIALS AND METHODS: Expression analysis, survival analysis and co-expression analysis of AHSG were performed using a public database, and cytological and molecular biology assays were performed to explore the role of AHSG in lung adenocarcinoma. RESULT: Compared with normal tissues, AHSG expression was significantly higher in cancer tissues in the TCGA-LUAD database, and pan-cancer analysis revealed abnormal AHSG expression in different kinds of tumours. Survival analysis revealed that compared with the low expression group, the patients in the high expression group had a significantly worse overall survival duration in the TCGA-LUAD database, and a subsequent study confirmed that AHSG expression could be an independent prognostic factor of overall survival in lung adenocarcinoma. AHSG-related genes are involved in multiple physiological and pathophysiological pathways. In subsequent cytological and molecular biology experiments, inhibition of AHSG expression suppressed proliferation, migration and invasion in lung adenocarcinoma cell lines, and the EMT process was blocked after knockdown of AHSG. CONCLUSION: AHSG could be used as a prognostic factor for OS in patients with lung adenocarcinoma. It can promote the biological behaviour of lung adenocarcinoma and may become a potential target for treatment, which is worthy of further study.

PKNOX2 suppresses lung cancer cell proliferation by inhibiting the PI3K/AKT/mTOR axis.

PBX/knotted 1 homeobox 2 (PKNOX2) has been implicated in tumorigenesis; however, its role in lung cancer (LC) remains unknown. The present study thus aimed to examine the expression, regulation, function and clinical implication of PKNOX2 in LC. A series of experiments were performed, including Cell Counting Kit-8 assay, cell cycle analysis, wound-healing assay, Transwell assay, methylation-specific PCR and western blotting. Bioinformatics analysis revealed that PKNOX2 was a LC-related gene, and a decrease in its expression was found in LC tissues from three public datasets. The results of reverse transcription-quantitative PCR assays also confirmed that PKNOX2 mRNA expression was markedly downregulated in LC tissues (n=60, P<0.01) and in five types of LC cell lines, and this was associated with the promoter methylation of PKNOX2. In addition, PKNOX2 expression was significantly associated with tumor invasion (P<0.0001), lymph node metastasis (P=0.0057) and TNM stage (P=0.0003); however, it was not associated with sex, age, pathological type or distant metastasis. The data obtained in vitro demonstrated that PKNOX2 silencing promoted LC cell proliferation and inhibited cell cycle arrest, accompanied by an increase in the expression levels of cell cycle-related proteins (cyclinD1, cyclinE1, CDK2 and CDK4), whereas PKNOX2 overexpression exhibited the opposite trend. In addition, PKNOX2 inhibited the migration and invasion of LC cells. Mechanistically, PKNOX2 knockdown activated the PI3K/AKT/mTOR signaling pathway by accelerating the phosphorylation of PI3K, AKT and mTOR, whereas PKNOX2 overexpression inactivated this signaling pathway. In conclusion, the findings of the present study suggested that PKNOX2 may suppress LC cell proliferation by inhibiting the PI3K/AKT/mTOR axis.

Soluble Pearl Extract provides effective skin lightening by antagonizing endothelin.

BACKGROUND: Incidence of skin pigmentation disorders has been on the rise globally. This calls for safer and more effective topical skin lightening and freckle-removing products. In this study, we hypothesized that Soluble Pearl Extract (SPE) may possess endothelin antagonizing compounds with good skin whitening effects. OBJECTIVES: (a) To determine the effect and mechanisms of SPE on ET-1-treated B16 melanoma cells. (b) To explore the cytotoxic effects of SPE on B16 melanoma cells. METHODS: CCK-8 assay was performed to determine how SPE and ET-1 affect the proliferation rate of B16 melanoma cells, the NaOH lysis assay was conducted to quantify the content of melanin while the tyrosinase activity was determined by DOPA oxidation test. The mRNA and protein expression levels of TYR and TRP-1 were determined by qRT-PCR assay and Western blot assay, respectively. RESULTS: We found that SPE at 0.1 and 1 µg/mL concentrations has no effect on the proliferation of the cells and 10 nmol/L ET-1 promoted B16 melanoma cells proliferation. Notably, B16 melanoma cells treated with 10 nmol/L ET-1 exhibited significantly higher melanin synthesis, tyrosinase activity, TYR, and TRP-1 mRNA expression levels compared with untreated cells. Of note, the effects of 10 nmol/L ET-1 treatment were abolished with SPE in a dose-dependent manner. CONCLUSIONS: SPE inhibits endothelin thereby safely and effectively lightening lightens the skin by antagonizing endothelin. Moreover, SPE is safe and effective.

Cardiac toxicity of Triptergium wilfordii Hook F. may correlate with its inhibition to hERG channel.

Tripterygium wilfordii Hook F. (TWHF) is a Chinese traditional medicine with cardiac toxicities. However, the mechanism of acute cardiac toxicity is not very clear. By using patch clamp techniques, we found that 0.05 mg/ml and 0.1 mg/ml of the aqueous crude extract of TWHF inhibit 21.4 ± 1.6% and 86.7 ± 5.7% (n = 5) of hERG current Amplitudes (IhERG) respectively. We further found that Celastrol, one of main components of TWHF, inhibits hERG with an IC50 of 0.83 µM. Additional mutagenesis studies show that mutations of T623A, S624A and F656A significantly alter the inhibition and S624A has the strongest effect, supported by our docking model. Our data suggest that inhibition of hERG channel activity by Celastrol contributed to TWHF cardiotoxicity.

[Effects of 17-DMAG on non-small cell lung cancer cell lines A549 and H1975
 being resistant to EGFR-TKI].

BACKGROUND AND OBJECTIVE: In the clinical treatment of patients with non-small cell lung cancer (NSCLC), the primary and acquired resistance of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) limits its clinical application, this need to explore new strategy or method to overcome this problem. Recently, some literatures have indicated that the antitumor role of heat shock protein 90 (HSP90) inhibitors by a variety of pathways may provide new strategy for resolving this problem. In this study, we examined the effect of 17-DMAG on NSCLC cell lines A549 and H1975 which were primary and acquired resistant to EGFR-TKI respectively, the purpose was to explore its influence on cell proliferation, apoptosis and the expression of EGFR in vitro as well as possible mechanism. METHODS: After A549 and H1975 cell lines were treated with different concentrations of 17-DMAG respectively, the inhibitory rate of cell proliferation was measured by MTT assay in 24 h, 48 h and 72 h. We investigated the effect of 17-DMAG on the cell apoptosis with flow cytometry and the expression of HSP90 and EGFR with Western blot after treated with 17-DMAG for 48 h. RESULTS: After treated with 17-DMAG, the inhibitory rate of different concentrations and time groups was significant (P<0.01), and the effect was in time- and dose-dependent manner; the apoptosis rate of both two cell lines in all treated groups were significantly higher than control group (P<0.01), and the effect was in dose-dependent manner. By Western blot analysis, there was no significant difference between all treated groups and control group for the expression of both HSP90 and EGFR protein in A549 cell line and HSP90 protein in H1975 cell line after exposed to 17-DMAG for 48 h (P>0.05), while the difference was significant for the expression of EGFR protein in H1975 cell line (P<0.01). CONCLUSIONS: 17-DMAG inhibited the proliferation of NSCLC cell lines A549 and H1975 and also induced apoptosis of both cell lines. It down-regulated the expression of mutant EGFR protein while this phenomenon was not observed in EGFR-wild type cell line. This suggested that the mechanism maybe different between A549 and H1975 cell lines with different genetic backgroud. Our study provided new strategy for treatment with NSCLC being resistant to EGFR-TKI.

[Effect of erythromycin on electrical activity and gastric emptying of the intrathoracic stomach after esophagectomy for esophageal cancer].

OBJECTIVE: To investigate the effect of erythromycin on electrical activity and emptying of the intrathoracic stomach after esophagectomy for esophageal cancer. METHODS: Thirty patients undergoing esophagectomy for cancer and esophagogastrostomy above the aortic arch were divided into the study group (n=15) and the control group (n=15). Electrogastrography and radionuclide gastric emptying were examined for these patients before and 1, 3, 6, 12 months after surgery. Patients in the study group received erythromycin (0.25 g tid po) for 1 week before examination. RESULTS: The wave amplitude (Uv), dominant frequency (CPM) and percentage of normal slow wave (%) of electrogastrogram decreased after surgery and returned to normal at the first postoperative month in the study group and the 12th postoperative month in the control group (P>0.05). Gastric emptying was significantly delayed after esophagectomy, and returned to normal one year after operation in the study group (P>0.05). However, gastric emptying remained abnormal in the control group (P<0.01). CONCLUSIONS: Erythromycin improves electrical activity and emptying of the stomach after esophagectomy for cancer. Gastric emptying recovery later than the recovery of electrical activity, which may be related to gastric ischemia and edema.

[Antireflux anastomosis following resection of esophageal cancer].

OBJECTIVE: To investigate the antireflux effects of a modified Nissen fundoplication following esophagectomy for cancer. METHODS: From March 2006 to March 2007, 70 patients with esophageal cancer were divided into two groups randomly. Esophagogastrostomy with a stapler only was perform in 35 patients as controls (group C), and a modified Nissen fundoplication was added after esophagogastrostomy with stapler in the other 35 patients as observed group (group O). There were 48 male and 22 female, ranging in age from 47 to 77 years (mean 60.1 years). The operative morbidity and mortality were recorded. Fourty-nine patients were followed at 3 months after surgery, and the questionnaire of life quality (EORTC QLQ C-30) was conducted in 24 patients in group C and 25 patients in group O. Thirty patients were examined with esophageal manometry, 24 h pH monitoring and gastroscopy. There were 16 patients in group C and 14 patients in group O. RESULTS: There was no significant difference in postoperative morbidity between the two groups (P > 0.05). However, the scores of heart burn and regurgitation in the group O were less than in group C (P = 0.041 and 0.034 respectively), but there was no difference in scores of dysphagia between the two groups (P = 0.677). The pressure at the anastomotic site was higher than that in the stomach in group O (P = 0.032), but not in group C (P = 0.448). DeMeester score in group O was 53 ± 46, compared to 140 ± 103 in group C (P = 0.043). The score of esophagitis was 0.9 ± 0.8 in group O, which was lower than 1.6 ± 1.0 in group C (P = 0.041). CONCLUSIONS: Addition of modified Nissen fundoplication after esophagectomy and esophagogastrostomy for cancer significantly increases the pressure at the anastomotic site, thus reduces the extent of gastroesophageal reflux, which leads to the reduction of the extent of reflux esophagitis and the improvement of the quality of life.

[Long-term outcome of antireflux surgery for gastroesophageal reflux disease].

OBJECTIVE: To investigate the long-term outcomes of various antireflux procedures for gastroesophageal reflux disease (GERD). METHODS: Between November 1988 and January 2004, 129 patients with GERD underwent antireflux procedures. Six kinds of antireflux procedures were performed including Nissen fundoplication, cardiac oblique invagination (COI) procedure, Belsey Mark IV, Toupet, Thal and Dor procedures. One hundred and sixteen patients were followed up. Esophageal manometry study was carried out in 95 patients preoperatively and 51 postoperatively. 24-hour esophageal pH monitoring were carried out in 56 patients preoperatively and 35 postoperatively. Esophagoscopy were performed in all patients before operation and 48 cases after operation. RESULTS: Clinical symptom scores reduced significantly from 4.1 +/- 0.4 before surgery to 1.1 +/- 1.0 after surgery (t = 27.21, P < 0.01). The outcome of surgery showed excellent in 42 cases (36.2%), good in 60 (51.7%), fair in 7 (6.0%), poor in 7 (6.0%). The long-term follow-up showed excellent or good results in 87.9% of patients. There was no significant difference in Nissen fundoplication, COI procedure and Belsey Mark IV. CONCLUSIONS: There are significant differences in symptom score, esophageal manometry, 24-hour esophageal pH monitoring and esophagoscopy pre- and post-operatively. There is no significant difference in Nissen fundoplication, COI procedure and Belsey Mark IV.

[Functional study of the remnant esophagus and intrathoracic stomach in patients who underwent esophagectomy for cancer].

OBJECTIVE: To investigate pathophysiological changes of the remnant esophagus and gastric cardia in patients who underwent esophagectomy for cancer, and to provide objective evidences for the improvement of the postoperative quality of life. METHODS: The function of the remnant esophagus and intrathoracic stomach in patients who underwent esophagectomy for cancer were assessed objectively. The methods that we used were gastric scintigraphy, esophageal manometry, 24-hour esophageal pH monitoring, electronic gastroscopy, videofluoroscopy, and DeMeester scoring system for the assessment of heartburn. Findings were recorded and compared with normal controls. RESULTS: After esophagectomy for cancer, the emptying of intrathoracic stomach was delayed (t = 7.105, P < 0.01) and improved over time, but could not reach normal one year after surgery (t = 2.9, P = 0.016). In patients who had undergone esophagectomy for cancer, the contracting pressure of the upper esophageal sphincter and resting pressure of the remnant esophagus were higher than that in normal controls (t = 2.275, P = 0.03; t = 2.16, P = 0.039 respectively). 89.7% of patients who had undergone esophagectomy had gastroesophageal reflux measured with 24-hour pH monitoring. The extent of reflux was less severe when patients were in a semi-reclining position than in a prostration position (t = 3.074, P = 0.005). CONCLUSION: After esophagectomy for cancer, delayed emptying of the intrathoracic stomach is improved gradually over time, but it is inaccessible to normal level. Gastroesophageal reflux extensively exists in patients who underwent esophagectomy for cancer, but it can be lessened by taking semi-reclining position.

[Expression and prognostic significance of survivin and caspase-3 in esophageal squamous-cell carcinoma and their relationship with HSPs expression].

OBJECTIVE: To explore the prognostic significance of expression of survivin and caspase-3 in esophageal squamous-cell carcinoma (ESCC) and the relasionship with expression of heat shock proteins 27 and 70 (HSP27 and HSP70). METHODS: Expressions of survivin and caspase-3 in 101 cases of ESCC were quantitatively detected with flow cytometry. Their expressions in long-term survival group (group A, >or= 5 years, 38 cases) were compared with those in the short-term survival group (group B, 0.05). The positive expression rate of survivin in group A was significantly lower than that in group B (31.6% vs 54.0%, P = 0.029). Compared with that in short-term survival group, the strong positive expression rate of caspase-3 in long-term survival group was significantly higher (47.6% vs. 68.4%, P = 0.042). Positive expression rate of caspase-3 showed decreasing tendency with increase in age. No significant differences in clinicopathologic features in relation to expression rate of caspase-3 other than tumor length. No correlation was observed between expression intensity of survivin and any clinicopathologic features. Logistic regression analysis indicated that survivin and caspase-3 expressions were of independent prognostic significance for ESCC. There was no association between survivin and caspase-3 expression and expression of HSP27 and HSP70. CONCLUSION: The expressions of survivin and caspase-3 are two independent prognostic factors in ESCC. They do not correlate with HSP27 and HSP70 expression.

[Prognostic significances of natural killer cells and dendritic cells infiltrations in esophageal squamous cell carcinoma].

BACKGROUND & OBJECTIVE: Natural killer (NK) cell and dendritic cell (DC) play important roles in anti-tumor immunity. Heat shock protein (HSP) is also involved in anti-tumor immune mechanisms. This study was to explore prognostic significances of NK cells and DCs infiltrations in esophageal squamous cell carcinoma (ESCC), and their relationships with expressions of HSP27 and HSP70. METHODS: NK cells, and DCs infiltrations were detected by SP immunohistochemistry in 101 specimens of ESCC. Among the relevant 101 patients, 38 were classified into long-term survival group (>/=5 years), and 63 into short-term survival group ( 0.05, t test]. NK cells and DCs infiltrations in ESCC had no significant correlation with clinicopathologic features, such as length of tumor, depth of invasion, histological grade, and lymph node metastasis, etc. (P > 0.05). Binary logistic regression analysis indicated that infiltration of NK cells was of prognostic significance for ESCC (P=0.001), while DCs was not (P=0.842). Infiltration of NK cells was positively correlated with infiltration of DCs (r=0.266, P=0.007). Both infiltrations of NK cells and DCs were positively correlated with expression of HSP27 (P=0.017, P=0.018), while no such correlation with expression of HSP70 was found (P > 0.05). CONCLUSIONS: Infiltration of NK cells is an independent prognostic factor of ESCC, and might be a prognostic biomarker of ESCC. Infiltration of DCs has no correlation with prognosis of ESCC. Infiltrations of NK cells and DCs may have internal correlation with the expression of HSP27.

Responses of human clasp and sling fibers to neuromimetics.

BACKGROUND AND AIMS: It has previously been demonstrated that clasp and sling fibers at the human gastroesophageal junction respond differently to acetylcholine (Ach). The present study was undertaken to investigate the differences between the physiological and pharmacological properties of the two types of muscle fiber. METHODS: Recordings were made of the isometric tension of human sling and clasp fibers in response to Ach, dopamine (DA), phenylephrine (Phe), and isoprenaline (Iso). These specimens were obtained from 18 patients who were operated on for esophageal cancer. RESULTS: Both Ach and Phe increased the tension of the two types of muscle; the values in the Ach group being 3-4-fold greater than those in the Phe group, while Iso decreased the tension of both types of muscle strip. The tension of the sling fibers was reduced by DA at lower concentration, and then increased gradually as the concentration was increased. In contrast, the tension of the clasp fibers did not obviously change when the concentration of DA was lower, but a slow elevation of tension was seen with the increase in DA concentration. CONCLUSIONS: The sensitivities and maximum responses to each agent differed between the clasp fibers and sling fibers. This suggests that the two kinds of fiber have different roles in establishing tension in the lower esophageal sphincter, with implications for the medical and surgical treatment of disorders in this region.

Long-term outcome of esophageal myotomy for achalasia.

AIM: Modified Heller's myotomy is still the first choice for achalasia and the assessment of surgical outcomes is usually made based on the subjective sensation of patients. This study was to objectively assess the long-term outcomes of esophageal myotomy for achalasia using esophageal manometry, 24-hour pH monitoring, esophageal scintigraphy and fiberoptic esophagoscopy. METHODS: From February 1979 to October 2000, 176 patients with achalasia underwent modified Heller's myotomy, including esophageal myotomy alone in 146 patients, myotomy in combination with Gallone or Dor antireflux procedure in 22 and 8 patients, respectively. Clinical score, pressure of the lower esophageal sphincter (LES), esophageal clearance rate and gastroesophageal reflux were determined before and 1 to 22 years after surgery. RESULTS: After a median follow-up of 14 years, 84.5% of patients had a good or excellent relief of symptoms, and clinical scores as well as resting pressures of the esophageal body and LES were reduced compared with preoperative values (P<0.001). However, there was no significant difference in DeMeester score between pre- and postoperative patients (P=0.51). Esophageal transit was improved in postoperative patients, but still slower than that in normal controls. The incidence of gastroesophageal reflux in patients who underwent esophageal myotomy alone was 63.6% compared to 27.3% in those who underwent myotomy and antireflux procedure (P=0.087). Three (1.7%) patients were complicated with esophageal cancer after surgery. CONCLUSION: Esophageal myotomy for achalasia can reduce the resting pressures of the esophageal body and LES and improve esophageal transit and dysphagia. Myotomy in combination with antireflux procedure can prevent gastroesophageal reflux to a certain extent, but further randomized studies should be carried out to demonstrate its efficacy.