Congener-specific fate and impact of microcystins in the soil-earthworm system.

Microcystins (MCs) have a significant influence on aquatic ecosystems, but little is known about their terrestrial fate and impact. Here, we investigated the fate of two MCs (MC-LR and MC-RR) in the soil-earthworm system, with consideration of their congener-specific impact on earthworm health, soil bacteria, and soil metabolome. Although MCs had little acute lethal effect on earthworms, they caused obvious growth inhibition and setae rupture. Relative to MC-RR, MC-LR exhibited higher bioaccumulation and the resulting dermal lesions and deformation of longitudinal muscles. While the incorporation of both MCs into soils stimulated pathogenic bacteria and depressed oxidative stress tolerant bacteria, the response among soil nitrification and glutathione metabolism differed between the two congeners. The dissipation kinetics of MCs obeyed the first-order model. Earthworms stimulated soil N-cycling enzyme activities, increased the abundance of MC-degrading bacteria, and promoted bacterial metabolic functions related to glutathione metabolism, xenobiotics biodegradation, and metabolism of amino acids that comprise MCs, which accelerated the dissipation of MC-LR and MC-RR by 227% and 82%, respectively. These results provide evidence of significant congener differences in the terrestrial fate and impact of MCs, which will enable a better understanding of their role in mediating soil functions and ecosystem services.

Analysis of clinicopathological features and prognostic factors of breast cancer brain metastasis.

BACKGROUND: Breast cancer (BC) has become the most common malignancy in women. The incidence and detection rates of BC brain metastasis (BCBM) have increased with the progress of imaging, multidisciplinary treatment techniques and the extension of survival time of BC patients. BM seriously affects the quality of life and sur-vival prognosis of BC patients. Therefore, clinical research on the clinicopathological features and prognostic factors of BCBM is valuable. By analyzing the clinicopathological parameters of BCBM patients, and assessing the risk factors and prognostic indicators, we can perform hierarchical diagnosis and treatment on the high-risk population of BCBM, and achieve clinical benefits of early diagnosis and treatment. AIM: To explore the clinicopathological features and prognostic factors of BCBM, and provide references for diagnosis, treatment and management of BCBM. METHODS: The clinicopathological data of 68 BCBM patients admitted to the Air Force Medical Center, Chinese People's Liberation Army (formerly Air Force General Hospital) from 2000 to 2022 were collected. Another 136 BC patients without BM were matched at a ratio of 1:2 based on the age and site of onset for retrospective analysis. Categorical data were subjected to χ2 test or Fisher's exact probability test, and the variables with P < 0.05 in the univariate Cox proportional hazards model were incorporated into the multivariate model to identify high-risk factors and independent prognostic factors of BCBM, with a hazard ratio (HR) > 1 suggesting poor prognostic factors. The survival time of patients was estimated by the Kaplan-Meier method, and overall survival was compared between groups by log-rank test. RESULTS: Multivariate Cox regression analysis showed that patients with stage III/IV tumor at initial diagnosis [HR: 5.58, 95% confidence interval (CI): 1.99-15.68], lung metastasis (HR: 24.18, 95%CI: 6.40-91.43), human epidermal growth factor receptor 2 (HER2)-overexpressing BC and triple-negative BC were more prone to BM. As can be seen from the prognostic data, 52 of the 68 BCBM patients had died by the end of follow-up, and the median time from diagnosis of BC to the occurrence of BM and from the occurrence of BM to death or last follow-up was 33.5 and 14 mo, respectively. It was confirmed by multivariate Cox regression analysis that patients with neurological symptoms (HR: 1.923, 95%CI: 1.005-3.680), with bone metastasis (HR: 2.011, 95%CI: 1.056-3.831), and BM of HER2-overexpressing and triple-negative BC had shorter survival time. CONCLUSION: HER2-overexpressing, triple-negative BC, late tumor stage and lung metastasis are risk factors of BM. The presence of neurological symptoms, bone metastasis, and molecular type are influencing prognosis factors of BCBM.

A prospective, single-centre, randomized, double-blind controlled study protocol to study whether long-term oral metronidazole can effectively reduce the incidence of postoperative liver metastasis in patients with colorectal cancer.

INTRODUCTION: Fifteen to 25% of patients with colorectal cancer have combined liver metastases at the time of diagnosis, whereas an additional 15 to 25% will develop liver metastases after curative resection of primary colorectal cancer, with the vast majority (80-90%) of liver metastases unresponsive to curative resection at first. Colorectal cancer liver metastasis is also the leading cause of death in patients with colorectal cancer. In recent years, several studies have demonstrated that intestinal flora, especially Fusobacterium nucleatum, plays a crucial role in the development of colorectal cancer liver metastasis, so we hypothesized that long-term metronidazole use could effectively reduce the incidence of postoperative liver metastasis in colorectal cancer patients. METHODS/DESIGN: This study is a prospective, single-centre, randomized, double-blind controlled study in which 300 patients will be randomly assigned to the test group or the control group in a 1:1 allocation ratio. The aim of this trial is to demonstrate that long-term oral antibiotics can effectively reduce the incidence of postoperative liver metastasis in patients with colorectal cancer. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Ethics Committee at the Chinese Ethics Committee of Registering Clinical Trials (ChiECRCT20210229). The results of this study will be disseminated at several research conferences and as published articles in peer-reviewed journals. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2100046201. Registered on July 05, 2021.

Peutz-Jeghers syndrome without STK11 mutation may correlate with less severe clinical manifestations in Chinese patients.

BACKGROUND: Peutz-Jeghers syndrome (PJS) is an autosomal dominant genetic disease with skin mucosal pigment spots and gastrointestinal (GI) multiple hamartoma polyps as clinical characteristics. At present, it is considered that the germline mutation of STK11 gene is the genetic cause of PJS. However, not all PJS patients can be detected STK11 germline mutations. The specific clinical characteristics of these PJS patients without STK11 mutation is an interesting clinical question. Or, like wild type GI stromal tumor, whether these PJS without STK11 mutation are also called PJS is worth discussing. Therefore, we designed the study to understand the clinical characteristics of these PJS patients without STK11 mutation. AIM: To investigates whether PJS patients with known STK11 mutations have a more severe spectrum of clinical phenotypes compared to those without. METHODS: A total of 92 patients with PJS admitted to the Air Force Medical Center from 2010 to 2022 were randomly selected for study. Genomic DNA samples were extracted from peripheral blood samples, and pathogenic germline mutations of STK11 were detected by high-throughput next-generation gene sequencing. Clinical-pathologic manifestations of patients with and without STK11/LKB1 mutations were compared. RESULTS: STK11 germline mutations were observed in 73 patients with PJS. Among 19 patients with no detectable STK11 mutations, six had no pathogenic germline mutations of other genes, while 13 had other genetic mutations. Compared with PJS patients with STK11 mutations, those without tended to be older at the age of initial treatment, age of first intussusception and age of initial surgery. They also had a lower number of total hospitalizations relating to intussusception or intestinal obstruction, and a lower load of small intestine polyps. CONCLUSION: PJS patients without STK11 mutations might have less severe clinical-pathologic manifestations than those with.

Clinical features, diagnosis, and treatment of Peutz-Jeghers syndrome: Experience with 566 Chinese cases.

BACKGROUND: Peutz-Jeghers syndrome (PJS) is a clinically rare disease with pigmented spots on the lips and mucous membranes and extremities, scattered gastrointestinal polyps, and susceptibility to tumors as clinical manifestations. Effective preventive and curative methods are still lacking. Here we summarize our experience with 566 Chinese patients with PJS from a Chinese medical center with regard to the clinical features, diagnosis, and treatment. AIM: To explore the clinical features, diagnosis, and treatment of PJS in a Chinese medical center. METHODS: The diagnosis and treatment information of 566 cases of PJS admitted to the Air Force Medical Center from January 1994 to October 2022 was summarized. A clinical database was established covering age, gender, ethnicity, family history, age at first treatment, time and sequence of appearance of mucocutaneous pigmentation, polyp distribution, quantity, and diameter, frequency of hospitalization, frequency of surgical operations, etc. The clinical data was retrospectively analyzed using SPSS 26.0 software, with P < 0.05 considered statistically significant. RESULTS: Of all the patients included, 55.3% were male and 44.7% were female. Median time to the appearance of mucocutaneous pigmentation was 2 years, and median time from the appearance of mucocutaneous pigmentation to the occurrence of abdominal symptoms was 10 years. The vast majority (92.2%) of patients underwent small bowel endoscopy and treatment, with 2.3% having serious complications. There was a statistically significant difference in the number of enteroscopies between patients with and without canceration (P = 0.004, Z = -2.882); 71.2% of patients underwent surgical operation, 75.6% of patients underwent surgical operation before the age of 35 years, and there was a statistically significant difference in the frequency of surgical operations between patients with and without cancer (P = 0.000, Z = -5.127). At 40 years of age, the cumulative risk of intussusception in PJS was approximately 72.0%, and at 50 years, the cumulative risk of intussusception in PJS was approximately 89.6%. At 50 years of age, the cumulative risk of cancer in PJS was approximately 49.3%, and at 60 years of age, the cumulative risk of cancer in PJS was approximately 71.7%. CONCLUSION: The risk of intussusception and cancer of PJS polyps increases with age. PJS patients ≥ 10 years old should undergo annual enteroscopy. Endoscopic treatment has a good safety profile and can reduce the occurrence of polyps intussusception and cancer. Surgery should be conducted to protect the gastrointestinal system by removing polyps.

Polyethylene and polypropylene microplastics reduce chemisorption of cadmium in paddy soil and increase its bioaccessibility and bioavailability.

Microplastics (MPs) usually coexist with heavy metals (HMs) in soil. MPs can influence HMs mobility and bioavailability, but the underlying mechanisms remain largely unexplored. Here, polyethylene and polypropylene MPs were selected to investigate their effects and mechanisms of sorption-desorption, bioaccessibility and bioavailability of cadmium (Cd) in paddy soil. Batch experiments indicated that MPs significantly reduced the Cd sorption in soil (p < 0.05). Accordingly, soil with the MPs had lower boundary diffusion constant of Cd (C1= 0.847∼1.020) and the Freundlich sorption constant (KF = 0.444-0.616) than that without the MPs (C1 = 0.894∼1.035, KF = 0.500-0.655). X-ray diffraction, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy analyses suggested that the MPs reduced Cd chemisorption, by covering the soil active sites and thus blocking complexation of Cd with active oxygen sites and interrupting the formation of CdCO3 and Cd3P2 precipitates. Such effects of MPs enhanced about 1.2-1.5 times of Cd bioaccessibility and bioavailability in soil. Almost the same effects but different mechanisms of polyethylene and polypropylene MPs on Cd sorption in the soil indicated the complexity and pervasiveness of their effects. The findings provide new insights into impacts of MPs on the fate and risk of HMs in agricultural soil.

Rno_circRNA_008646 regulates formaldehyde induced lung injury through Rno-miR-224 mediated FOXI1/CFTR axis.

Formaldehyde (FA) serves as a prevailing air pollutant, which has seriously threatened public health in recent years. Of all the known health effects, lung injury is one of the most severe risks. However, little is known about the circRNAs related molecular mechanism in the development of lung injury induced by FA. This study was designed to explore the potential roles of dysregulated circRNAs as well as its mechanism in FA-induced lung injury. In the present study, 24 male SD rats were exposed to formaldehyde (control, 0.5, 2.46 and 5 mg/m3) for 8 h per day for 8 weeks to induce lung injury. We used H&E staining to evaluate the histopathological changes of lung injury indifferent groups. The expression of circRNAs in lung tissue was detected by real-time PCR. Meanwhile, circRNA/miRNA/mRNA interaction networks were predicted by bioinformatics analysis. Our study revealed that formaldehyde exposure resulted in abnormal histopathological changes in lung tissues. Moreover, the expression of rno_circRNA_008646 was significantly higher in lung tissues of formaldehyde exposure rats than in control. Bioinformatics analysis showed that one potential target miRNA/mRNA for rno_circRNA_008646 was rno-miR-224/Forkhead Box I1 (FOXI1). Besides, luciferase report gene confirmed that there was targeted binding relationship between rno_circRNA_008646 and rno-miR-224, rno-miR-224 and FOXI1. Further verification experiments indicated that the expression of rno_circRNA_008646 was negatively correlated rno-miR-224, while it was positively correlated with FOXI1. JASPAR database showed transcription factor FOXI1 located in promotor of CF Transmembrane Conductance Regulator (CFTR). Both FOXI1 and CFTR were up-regulated in lung tissues after formaldehyde exposure. In conclusion, our findings suggested that formaldehyde may induce lung injury, and this may be caused by up-regulatedrno_circRNA_008646, which medicated rno-miR-224/FOXI1/CFTR axis.

Protocol for Comparing the Efficacy of Three Reconstruction Methods of the Digestive Tract (Kamikawa Versus Double-Tract Reconstruction Versus Tube-Like Stomach) After Proximal Gastrectomy.

Background: Appropriate gastrointestinal reconstruction after proximal gastrectomy can effectively reduce the incidence of postoperative complications in patients with proximal early gastric cancer. However, there is still great controversy about the choice of digestive tract reconstruction after proximal gastrectomy, and there is no clinical consensus on the choice of digestive tract reconstruction after proximal gastrectomy. Currently, there is a lack of large-sample, prospective, randomized controlled studies to compare the efficacy of Kamikawa, double-tract reconstruction, and tube-like stomach reconstruction after proximal gastrectomy. Methods/design: This study will investigate the efficacy of three reconstruction methods after proximal gastrectomy in a prospective, multicenter, randomized controlled trial, which will enroll 180 patients with proximal early gastric cancer. Patients will be randomly divided into three groups: Group A (Kamikawa, n = 60), Group B (double-tract reconstruction, n = 60), and Group C (tube-like stomach, n = 60). The general information, past medical history, laboratory findings, imaging findings, and surgical procedures of the patients will be recorded and analyzed. The incidence of reflux esophagitis will be recorded as the primary endpoint. The incidence of anastomotic leakage, anastomotic stenosis, operative time and intraoperative blood loss will be recorded as secondary endpoints. Discussion: This study will establish a large-sample, prospective, randomized controlled trial to compare the efficacy of Kamikawa, double-tract reconstruction, and tube-like stomach reconstruction after proximal gastrectomy. Trial registration: This study was approved by the Chinese Clinical Trial Registry and registered on April 30, 2021. The registration number is ChiCTR2100045975.

A retrospective study of the role of preoperative ultrasonography in the detection of deep vein thrombosis in 1750 patients with gastric and colorectal cancers.

BACKGROUND: There has been a lack of research in the past on the prevalence and risk factors associated with deep vein thrombosis (DVT) in patients with resectable gastric and colorectal cancers. The purpose of this study was to review the anatomical distribution, prevalence and risk factors associated with lower limb DVT in 1750 patients with preoperative gastric and colorectal cancers and to evaluate the role of preoperative ultrasonography in the detection of DVT in preventing postoperative pulmonary thromboembolism (PTE) in patients with gastric and colorectal cancers. METHODS: A total of 1750 patients with gastric and colorectal cancers who underwent preoperative venous ultrasonography of the lower limbs were retrospectively reviewed. The risk factors associated with preoperative DVT were identified using univariate and multivariate logistic regression analysis. RESULTS: Seventy-three of the 1750 patients with gastric and colorectal cancers had DVT detected by preoperative venous ultrasonography of the lower limb and the incidence of lower limb DVT was 4.17 % in 1750 patients with gastric and colorectal cancers. Univariate analysis showed a higher risk of DVT in patients who met the following criteria: aged ≥80 years, female sex, the performance status ≥1, stage IV, ASA class ≥ III/IV, and hypertension. Multivariate logistic regression analysis showed that female sex, stage IV and ASA class ≥ III/IV were significantly associated with DVT before gastric and colorectal cancer surgery. CONCLUSIONS: Our study showed that female sex, stage IV and ASA class ≥ III/IV were significantly associated with DVT before gastric and colorectal cancer surgery. Routine venous ultrasonography for the lower limb can identify the risk of PTE, which is of great significance in the prevention and occurrence of PTE.

Are preoperative oral antibiotics effective in reducing the incidence of anastomotic leakage after colorectal cancer surgery? Study protocol for a prospective, multicentre, randomized controlled study.

INTRODUCTION: The optimal preoperative preparation for elective colorectal cancer surgery has been debated in academic circles for decades. Previously, several expert teams have conducted studies on whether preoperative mechanical bowel preparation and oral antibiotics can effectively reduce the incidence of postoperative complications, such as surgical site infections and anastomotic leakage. Most of the results of these studies have suggested that preoperative mechanical bowel preparation for elective colon surgery has no significant effect on the occurrence of surgical site infections and anastomotic leakage. METHODS/DESIGN: This study will examine whether oral antibiotic bowel preparation (OABP) influences the incidence of anastomotic leakage after surgery in a prospective, multicentre, randomized controlled trial that will enrol 1500 patients who require colon surgery. The primary endpoint, incidence of anastomotic leakage, is based on 2.3% in the OABP ± mechanical bowel preparation (MBP) group in the study by Morris et al. Patients will be randomized (1:1) into two groups: the test group will be given antibiotics (both neomycin 1 g and metronidazole 0.9 g) the day before surgery, and the control group will not receive any special intestinal preparation before surgery, including oral antibiotics or mechanical intestinal preparation. All study-related clinical data, such as general patient information, past medical history, laboratory examination, imaging results, and surgery details, will be recorded before surgery and during the time of hospitalization. The occurrence of postoperative fistulas, including anastomotic leakage, will be recorded as the main severe postoperative adverse event and will represent the primary endpoint. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Chinese Ethics Committee of Registering Clinical Trials (ChiECRCT20200173). The results of this study will be disseminated at several research conferences and as published articles in peer-reviewed journals. Protocol was revised on November 22, 2021, version 4.0. TRIAL REGISTRATION: ChiCTR2000035550 . Registered on 13 Aug 2020.

The role of KMT2 gene in human tumors.

Histone methylation plays a crucial role in the regulation of gene transcriptional expression, and aberration of methylation-modifying enzyme genes can lead to a variety of genetic diseases, including human cancers. The histone modified protein KMT2 (lysin methyltransferase) family are involved in cell proliferation, growth, development and differentiation through regulating gene expression, and are closely related with many blood cancers and solid tumors. In recent years, several studies have shown that mutations in the KMT2 gene occur frequently in a variety of human cancers and the mutation status of the KMT2 gene may be correlated with the occurrence, development and prognosis of some tumors. Research uncovering the clinical characteristics and molecular mechanisms of KMT2 mutation in human tumors will be helpful for early diagnosis and prognosis of tumors as well as drug development for targeted therapies.

Diffuse invasive signet ring cell carcinoma in total colorectum caused by ulcerative colitis: A case report and review of literature.

BACKGROUND: Diffuse invasive signet ring cell carcinoma of the colorectum is extremely rare clinically. This type of colorectal cancer has certain clinical, pathological and biological characteristics that are different from ordinary colorectal cancer. CASE SUMMARY: A 31-year-old young woman was admitted to the hospital for nearly 1 wk due to recurrent symptoms of mucopurulent bloody stools and abdominal distension. Preoperative colonoscopy showed a ring-shaped intestinal wall mass 10 cm from the rectum to the anus. Three pieces of tumor tissue were removed for examination. The pathological results showed rectal mucinous adenocarcinoma. The patient underwent laparoscopic exploration under general anesthesia, and then laparoscopic total colorectal resection, ileal pouch-anal anastomosis and ileostomy were performed. The patient was switched to a FOLFOX + cetuximab regimen. After the fifth cycle, the patient was unable to tolerate further treatment due to tumor progression and multiple organ dysfunction, and died at the end of May 2020. Overall survival was 7 mo. CONCLUSION: Carcinogenesis of ulcerative colitis is different from sporadic colon cancer, and the overall prognosis is extremely poor.

Protective effect of mesenchymal stem cell-derived exosomal treatment of hippocampal neurons against oxygen-glucose deprivation/reperfusion-induced injury.

BACKGROUND: Individuals who survive a cardiac arrest often sustain cognitive impairments due to ischemia-reperfusion injury. Mesenchymal stem cell (MSC) transplantation is used to reduce tissue damage, but exosomes are more stable and highly conserved than MSCs. This study was conducted to investigate the therapeutic effects of MSC-derived exosomes (MSC-Exo) on cerebral ischemia-reperfusion injury in an in vitro model of oxygen-glucose deprivation/reperfusion (OGD/R), and to explore the underlying mechanisms. METHODS: Primary hippocampal neurons obtained from 18-day Sprague-Dawley rat embryos were subjected to OGD/R treatment, with or without MSC-Exo treatment. Exosomal integration, cell viability, mitochondrial membrane potential, and generation of reactive oxygen species (ROS) were examined. Terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick-end labeling (TUNEL) staining was performed to detect neuronal apoptosis. Moreover, mitochondrial function-associated gene expression, Nrf2 translocation, and expression of downstream antioxidant proteins were determined. RESULTS: MSC-Exo attenuated OGD/R-induced neuronal apoptosis and decreased ROS generation (P<0.05). The exosomes reduced OGD/R-induced Nrf2 translocation into the nucleus (2.14±0.65 vs. 5.48±1.09, P<0.01) and increased the intracellular expression of antioxidative proteins, including superoxide dismutase and glutathione peroxidase (17.18±0.97 vs. 14.40±0.62, and 20.65±2.23 vs. 16.44±2.05, respectively; P<0.05 for both). OGD/R significantly impaired the mitochondrial membrane potential and modulated the expression of mitochondrial function-associated genes, such as PINK, DJ1, LRRK2, Mfn-1, Mfn-2, and OPA1. The abovementioned changes were partially reversed by exosomal treatment of the hippocampal neurons. CONCLUSIONS: MSC-Exo treatment can alleviate OGD/R-induced oxidative stress and dysregulation of mitochondrial function-associated genes in hippocampal neurons. Therefore, MSC-Exo might be a potential therapeutic strategy to prevent OGD/R-induced neuronal injury.

Detection and analysis of common pathogenic germline mutations in Peutz-Jeghers syndrome.

BACKGROUND: Different types of pathogenic mutations may produce different clinical phenotypes, but a correlation between Peutz-Jeghers syndrome (PJS) genotype and clinical phenotype has not been found. Not all patients with PJS have detectable mutations of the STK11/LKB1 gene, what is the genetic basis of clinical phenotypic heterogeneity of PJS? Do PJS cases without STK11/LKB1 mutations have other pathogenic genes? Those are clinical problems that perplex doctors. AIM: The aim was to investigate the specific gene mutation of PJS, and the correlation between the genotype and clinical phenotype of PJS. METHODS: A total of 24 patients with PJS admitted to the Air Force Medical Center, PLA (formerly the Air Force General Hospital, PLA) from November 1994 to January 2020 were randomly selected for inclusion in the study. One hundred thirty-nine common hereditary tumor-related genes including STK11/LKB1 were screened and analyzed for pathogenic germline mutations by high-throughput next-generation sequencing (NGS). The mutation status of the genes and their relationship with clinical phenotypes of PJS were explored. RESULTS: Twenty of the 24 PJS patients in this group (83.3%) had STK11/LKB1 gene mutations, 90% of which were pathogenic mutations, and ten had new mutation sites. Pathogenic mutations in exon 7 of STK11/LKB1 gene were significantly lower than in other exons. Truncation mutations are more common in exons 1 and 4 of STK11/LKB1, and their pathogenicity was significantly higher than that of missense mutations. We also found SLX4 gene mutations in PJS patients. CONCLUSION: PJS has a relatively complicated genetic background. Changes in the sites responsible for coding functional proteins in exon 1 and exon 4 of STK11/LKB1 may be one of the main causes of PJS. Mutation of the SLX4 gene may be a cause of genetic heterogeneity in PJS.

[A 5 year follow-up study on non-surgical treatment of ruptured lumbar disc herniation].

OBJECTIVE: To investigate the short-term and mid-term efficacy with non-surgical treatment and to predict the long-term outcomes of ruptured lumbar disc herniation. METHODS: From February 2011 to February 2014, 75 patients with single-segment ruptured lumbar disc herniation treated by non-surgical therapy were selected for prospective study. There were 53 males and 22 females, aged from 18 to 58 (35.62±9.96) years old. The course of disease was from 5 days to 6 months, with an average of (46.45±40.66) days. The lesions were located at L3,4 in 4 cases, at L4,5 in 29 cases, at L5S1 in 42 cases. Radiation pain in 46 cases on the left and 29 cases on the right. The JOA score, straight leg raising test angle and finger-to-ground distance were assessed pretreatment and at 3 months, 6 months, 1 year, 2 years and 5 years after treatment. The improvement rate of JOA at the final follow up (5 years after treatment) was calculated and the curative effect according to JOA score was evaluated;the volume change of protrusion before treatment and at the final follow-up (5 years after treatment) was analyzed, the volume absorption rate of protrusion was calculated and the absorption of protrusion was observed;the relationship between the improvement rate of JOA and the absorption rate of protrusion were analyzed. RESULTS: Seventy one patients were finally followed up at 5 years after non surgical treatment. The JOA score, straight leg raising test angle and finger to ground distance at 3 months, 6 months, 1 year, 2 years and 5 years after treatment were significantly improved (P<0.05). There was no significant difference in JOA scores between 5 years and 6 months after treatment, 5 and 2 years after treatment, and 2 years and 6 months after treatment (P>0.05). There was significant differencein other time points (P<0.05). The results of the straight leg raising test angle and the finger-to-ground distance were similar to the JOA scores. The improvement rate of JOA score was (62.69± 2.47)% at the final follow-up. According to JOA score, the results were excellent in 26 cases, good in 26 cases, fair in 14 cases, poor in 5 cases, and the excellent and good rate was 73.24%. The volume of protrusion decreased from (1 981.73±588.72) mm3 to (1 011.82±395.47) mm3, the total absorption rate was (45.65±2.83)%, the protrusion was obviously absorbed in 24 cases, partially absorbed in 26 cases, not absorbed in 19 cases, and increased in 2 cases. It was found that there was a positive correlation between improvement rate of JOA scores and protrusion absorption rate at 5 years after non surgical treatment (r= 0.679, P<0.001). CONCLUSION: Non-surgical treatment of ruptured lumbar disc herniation can achieve good results, clear the characteristics of the ruptured lumbar disc herniation and prognosis, and some patients have "reabsorption" phenomenon.

Mutation analysis of related genes in hamartoma polyp tissue of Peutz-Jeghers syndrome.

BACKGROUND: Peutz-Jeghers syndrome (PJS) is a rare disease with clinical manifestations of pigmented spots on the lips, mucous membranes and extremities, scattered gastrointestinal polyps, and susceptibility to tumors. The clinical heterogeneity of PJS is obvious, and the relationship between clinical phenotype and genotype is still unclear. AIM: To investigate the mutation status of hereditary colorectal tumor-associated genes in hamartoma polyp tissue of PJS patients and discuss its relationship with the clinicopathological data of PJS. METHODS: Twenty patients with PJS were randomly selected for this study and were treated in the Air Force Medical Center (former Air Force General Hospital) PLA between 2008 and 2017. Their hamartoma polyp tissues were used for APC, AXIN2, BMPR1A, EPCAM, MLH1, MLH3, MSH2, MSH6, MUTYH, PMS1, PMS2, PTEN, SMAD4, and LKB1/STK11 gene sequencing using next-generation sequencing technology. The correlations between the sequencing results and clinical pathological data of PJS were analyzed. RESULTS: Fourteen types of LKB1/STK11 mutations were detected in 16 cases (80.0%), of which 8 new mutations were found (3 types of frameshift deletion mutations: c.243delG, c.363_364delGA, and c.722delC; 2 types of frameshift insertions: c. 144_145insGCAAG, and c.454_455insC; 3 types of splice site mutations: c.464+1G>T, c.464+1G>A, and c.598-1G>A); 9 cases (45.0%) were found to have 18 types of heterozygous mutations in the remaining 13 genes except LKB1/STK11. Of these, MSH2: c.792+1G>A, MSH6: c.3689C>G, c.4001+13C>CTTAC, PMS1: c.46C>t, and c.922G>A were new mutations. CONCLUSION: The genetic mutations in hamartoma polyp tissue of PJS are complex and diverse. Moreover, other gene mutations in PJS hamartoma polyp tissue were observed, with the exception of LKB1/STK11 gene, especially the DNA mismatch repair gene (MMR). Colorectal hamartoma polyps with LKB1/STK11 mutations were larger in diameter than those with other gene mutations.

[Experimental study on the inhibition of Formononetin on the differentiation of osteoclasts induced by RANKL].

OBJECTIVE: To establish the in vitro study model of osteoclasts induced by RANKL, to elaborate the effect of formononetin (FO) , an effective component of Caulis Spatholobi, on the differentiation and function of bone marrow mononuclear macrophages (BMMs) into osteoclasts, and to explore the molecular mechanism of its inhibition. METHODS: The BMMs in femur and tibia were isolated from 20 clean C57/BL6 mice of 4 to 6 weeks old, 10 males and 10 females, each weighing (20± 2) g. The BMMs in femur and tibia were cultured and proliferated in vitro with α-MEM medium. BMMs were cultured with MCSF and different concentrations of anthocyanin (5 to 50 µm) respectively for 4 days, and CCK8 of cell proliferation and toxicity was detected. BMMs in good growth condition were added to M-CSF and RANKL to induce osteoclast differentiation in turn. There was no special treatment in the control group. DMSO was added to the control group with DMSO solvent. Each observation group was added with different concentrations of awnasin (1 to 20 µm) . After 6 days of culture, the osteoclasts were counted and statistically analyzed. The expression of NFATc1, c-Fos and ERK, the key transcription factors in osteoclast differentiation, were detected by Western blot, RNA was extracted at 4 days, and the activity of ctsk, trap, MMP9 and Car2 were detected by real time PCR. RESULTS: CCK8 test results showed that awnstein could inhibit the activity of BMMs in a dose-dependent manner, and had no significant toxic effect on the growth of bmms within the safe concentration range of ≤20 µM (P= 0.278>0.05) . The results of trap staining showed that awnstein could inhibit osteoclast production in a dose-dependent manner in the concentration range of (1 to 20 µM) , especially in 10 µM (P=0.000<0.05) . Western blot showed that 10 µ m could significantly inhibit the expression of NFATc1 and c-Fos, but not the expression of ERK. In terms of osteoclast function, the expression of ctsk (P=0.000<0.05) , trap (P=0.000<0.05) , MMP9 (P=0.000<0.05) and Car2 (P=0.000<0.05) related to osteoclast function were detected by real time PCR. CONCLUSION: The effective component of Caulis Spatholobi can inhibit the proliferation and differentiation of primary mononuclear macrophages into osteoclasts, and down regulate the expression of osteoclast bone resorption related proteins and genes, which may be one of the mechanisms of its prevention and treatment of bone destruction and collapse in osteonecrosis of femoral head.

Evaluation of D2-plus radical resection for gastric cancer with pyloric invasion.

BACKGROUND: The optimal lymphadenectomy for gastric cancer (GC) with pyloric invasion is controversial because the pattern of lymph node metastasis is different from that of distal GC. The rate of lymph node metastasis into the posterior area of the pancreatic head and hepatoduodenal ligament is high. This study evaluated the estimated benefit of radical gastrectomy with D2-plus lymphadenectomy in patients with pyloric invasion. METHODS: All patients with GC invading the pylorus who underwent curative surgical resection with D2-plus lymphadenectomy between February 2013 and September 2015 were enrolled in the study. The index of estimated benefit from lymph node dissection (IEBLD) was calculated by multiplying the incidence of metastasis to each lymph node station by the 3-year overall survival (OS) rate of patients with metastasis to that station. RESULTS: In total, 128 patients were eligible. The rate of lymph node metastasis and the 3-year OS rate (and IEBLD) of the patients with metastasis to lymph nodes were 14.3 and 44.4% (5.56) for No. 8p, 10.9 and 35.7% (3.89) for No. 12b, 9.5 and 33.3% (3.13) for No. 12p, 18.8 and 54.2% (10.19) for No. 13, and 21.8 and 53.6% (11.68) for No. 14v, respectively. CONCLUSIONS: In radical gastrectomy for GC with pyloric invasion, some survival benefit was observed with dissection of the No. 13 and No. 14 lymph nodes, but there was no survival benefit with dissection of the No. 8p lymph nodes. The No. 12b and No. 12p lymph nodes may be better to dissect in cT3 GC patients with pyloric invasion. TRIAL REGISTRATION: http://ClinicalTrials.gov Identifier: NCT01836991. Date of registration: April 17, 2013.

Novel abdominal approach for dissection of advanced type II/III adenocarcinoma of the esophagogastric junction: a new surgical option.

OBJECTIVE: The optimal surgical approach for Siewert type II adenocarcinoma of the esophagogastric junction (AEG) is controversial. In this study, we evaluated the outcomes of total gastrectomy for Siewert type II/III AEG via the left thoracic surgical approach that is used at our center. METHODS: We identified 41 patients with advanced AEG in our retrospective database and analyzed their 3-year survival rate, upper surgical margin, postoperative complications, and index of estimated benefit from lymph node dissection. RESULTS: The 3-year overall survival rate of the whole group was 63%, but no difference was observed between Siewert type II and III AEGs. Esophageal exposure and lymphadenectomy were sufficient. Eight patients developed postoperative complications, but none of the patients developed anastomotic leakage. Dissection of lymph node station Nos. 19 and 110 may be necessary for patients with Siewert type II AEG. Multivariate analysis revealed that the cT category was the only independent risk factor. CONCLUSIONS: Total gastrectomy via an approach from the abdominal cavity into the thoracic cavity may be an optimal surgical technique for advanced Siewert type II AEG.

Must Peutz-Jeghers syndrome patients have the LKB1/STK11 gene mutation? A case report and review of the literature.

Peutz-Jeghers syndrome (PJS) is an autosomal dominant inherited disease, which is characterized by mucocutaneous pigmentation and multiple gastrointestinal hamartoma polyps. The germline mutation of LKB1/STK11 gene on chromosome 19p13.3 is considered to be the hereditary cause of PJS. However, must a patient with PJS have the LKB1/STK11 gene mutation? We here report a case of a male patient who had typical manifestations of PJS and a definite family history, but did not have LKB1/STK11 gene mutation. By means of high-throughput sequencing technology, only mutations in APC gene (c.6662T > C: p.Met2221Thr) and MSH6 gene (c.3488A > T: p.Glu1163Val) were detected. The missense mutations in APC and MSH6 gene may lead to abnormalities in structure and function of their expression products, and may result in the occurrence of PJS. This study suggests that some other genetic disorders may cause PJS besides LKB1/STK11 gene mutation.