MiR-145 is upregulated in the retarded preimplantation embryos and modulates cholesterol levels in mice preimplantation embryos through targeting Abca1.

BACKGROUND: Preimplantation embryonic lethality is a driver of female infertility. Certain microRNAs (miRNAs) have previously been demonstrated to play important roles in the regulation of embryogenesis. METHODS: Normally developing blastocysts and arrested embryos were collected from patients undergoing intracytoplasmic sperm injection (ICSI), and the expression of specific miRNAs therein was evaluated by qPCR. The overexpression of target molecule miR-145 in early mice embryos was achieved via oocyte microinjection, enabling the subsequent monitoring of how such overexpression impacted embryonic development. Bioinformatics approaches were utilized to identify putative miR-145 target mRNAs, and luciferase reporter assessments were implemented to confirm the ability of miR-145 to regulate Abca1 in HEK293T cells. The functional relationship between miR-145 and Abca1 in the mice's embryonic development was then confirmed through rescue assays. RESULTS: Abnormally increased miR-145 expression was observed in patients' arrested embryos, and the exogenous overexpression of this miRNA significantly suppressed mural blastocyst formation. Mechanistically, miR-145 was found to bind to the 3'-untranslated area of the Abca1 mRNA in HK293T cells, thus suppressing its expression and increasing embryonic cholesterol levels. In line with the importance of these cholesterol levels to embryogenesis, the upregulation of Abca1 was sufficient to rescue the observed change in cholesterol levels and the associated retardation of mice embryonic development that occurred in response to the overexpression of miR-145. CONCLUSION: The regulatory dynamics of the miR-145/Abca1 axis play an important role in shaping normal embryonic development.

Analysis of risk factors leading to postoperative urethral stricture and bladder neck contracture following transurethral resection of prostate.

PURPOSE: To determine risk factors of postoperative urethral stricture (US) and vesical neck contracture (BNC) after transurethral resection of prostate (TURP) from perioperative parameters. MATERIALS AND METHODS: 373 patients underwent TURP in a Chinese center for lower urinary tract symptoms suggestive of benign prostatic obstruction (LUTS/BPO), with their perioperative and follow-up clinical data being collected. Univariate analyses were used to determine variables which had correlation with the incidence of US and BNC before logistic regression being applied to find out independent risk factors. RESULTS: The median follow-up was 29.3 months with the incidence of US and BNC being 7.8% and 5.4% respectively. Resection speed, reduction in hemoglobin (ΔHb) and hematocrit (ΔHCT) levels, incidence of urethral mucosa rupture, re-catheterization and continuous infection had significant correlation with US, while PSA level, storage score, total prostate volume (TPV), transitional zone volume (TZV), transitional zone index (TZI), resection time and resected gland weight had significant correlation with BNC. Lower resection speed (OR=0.48), urethral mucosa rupture (OR=2.44) and continuous infection (OR=1.49) as well as higher storage score (OR=2.51) and lower TPV (OR=0.15) were found to be the independent risk factors of US and BNC respectively. CONCLUSIONS: Lower resection speed, intraoperative urethral mucosa rupture and postoperative continuous infection were associated with a higher risk of US while severer storage phase symptom and smaller prostate size were associated with a higher risk of BNC after TURP.

Regulation of T cells in the tumor microenvironment by histone methylation: LSD1 inhibition-a new direction for enhancing immunotherapy.

Although immune checkpoint blockade (ICB) has been shown to achieve durable therapeutic responses in various types of tumors, only 20-40 % of patients benefit from this therapy. A growing body of research suggests that epigenetic modulation of the tumor microenvironment may be a promising direction for enhancing the efficacy of immunotherapy, for example, histone methylation plays an important role in the regulation of T cells in the tumor microenvironment (TME). In particular, histone lysine-specific demethylase 1 (LSD1/KDM1A), as an important histone-modifying enzyme in epigenetics, was found to be an important factor in the regulation of T cells. Therefore, this paper will summarize the effects of histone methylation, especially LSD1, on T cells in the TME to enhance the efficacy of anti-PD-1 immunotherapy. To provide a strong theoretical basis for the strategy of combining LSD1 inhibitors with anti-PD-1/PD-L1 immunotherapy, thus adding new possibilities to improve the survival of tumor patients.

Positive expression of LSD1 and negative expression of E-cadherin correlate with metastasis and poor prognosis of colon cancer.

BACKGROUND: The first identified lysine-specific demethylase, LSD1, plays an important role in the metastatic progression of several types of cancer. AIMS: The aim of this study was to investigate LSD1, E-cadherin, and N-cadherin expression in colon cancer specimens and their clinical significance. METHODS: The expression of LSD1, E-cadherin, and N-cadherin in colon cancer specimens was determined by immunohistochemistry, and the relationship between the expression of the respective molecules and clinicopathological characteristics was analyzed. RESULTS: The positive expression rates of LSD1, E-cadherin, and N-cadherin in colon cancer specimens were 66.7 % (72/108), 85.2 % (92/108), and 41.7 % (45/108), respectively. LSD1 was significantly more highly expressed in colon cancer specimens classified as high TNM stage lesions and with distant metastasis (P < 0.05). Further analysis demonstrated that LSD1 expression was positively correlated with lymph node and distant metastases (P < 0.05). However, E-cadherin expression was significantly downregulated in colon cancer specimens classified as high TNM stage lesions and with distant metastasis (P < 0.05), whereas the expression of N-cadherin did not differ significantly according to clinical and pathological characteristics (P > 0.05). Correlation analysis revealed that LSD1 expression was negatively correlated with E-cadherin expression (r s = -0.318, P = 0.001), but not evidently correlated with N-cadherin expression (r s = 0.182, P = 0.06). Colon cancer specimens with positive LSD1 expression and negative E-cadherin expression were correlated with significantly lower overall survival. CONCLUSIONS: LSD1 showed a significantly higher expression, in contrast to the significantly lower expression of E-cadherin, in colon cancer specimens classified as high TNM stage lesions and with distant metastasis. Positive expression of LSD1 and negative expression of E-cadherin may be predictors of a worse colon cancer prognosis.

Prevalence and characteristics of cervical human papillomavirus genotypes and cervical lesions among 58630 women from Guangzhou, China.

BACKGROUND: To assess the prevalence and characteristics of human papillomavirus (HPV) genotypes and its associated cervical lesions in Guangzhou, China, which may be useful for adjusting area-specific cervical cancer prevention and control strategies. METHODS: A total of 58630 women were enrolled. Cervical specimens were collected for HPV DNA testing and/or cervical cytology. Patients with visible cervical lesions or abnormal screening results underwent further cervical biopsies. RESULT: The overall HPV positive rate was 14.07%. The top five genotypes in Guangzhou were HPV 52 (3.06%), HPV 16 (2.28%), HPV 58 (1.80%), HPV 51 (1.32%), and HPV 39 (1.15%). The prevalence of overall HPV and vaccine-targeted HPV genotypes showed a significantly decreasing trend from 2016 to 2019 (P < 0.05). While, the infection rate of HPV 35 increased significantly during this time (P = 0.015). The age-specific prevalence of any HPV genotypes showed a bimodal curve, which peaked firstly among the < 20 y age group, and then peaked secondly among the > 59 y age group. Among HPV-positive women, the proportions of HSIL and cervical cancer increased significantly with age (P < 0.05). Among > 59 y age group, 9.35% HPV-positive cases were diagnosed as cervical cancer. HPV 16/18 was the most common cause of cervical cancer. While, the percentage of non-HPV 16/18 infection among cervical cancer patients increased over time, from 15.21% in 2015 to 26.32% in 2019 (P = 0.010). Besides that, the prevalence of non-HPV 16/18 genotypes among cervical cancer patients significantly increased with age, which peaked at the > 59 y age group (P = 0.014). CONCLUSION: Although the prevalence of any HPV and vaccine-targeted HPV genotypes decreased significantly with time, it is still important to follow the HPV genotypes and their associated cancer risk after the large-scale popularization of HPV vaccine. And age should be taken into consideration in screening strategies and risk-based management of cervical cancer in Guangzhou.

N-acetylcysteine promotes the proliferation of porcine adipose-derived stem cells during in vitro long-term expansion for cultured meat production.

Cultured meat is an efficient, safe and sustainable meat production technology. Adipose-derived stem cell (ADSC) is a promising cell type for cultured meat. In vitro, obtaining numerous of ADSCs is a pivotal step for cultured meat. In this research, we demonstrated that the proliferation and adipogenic differentiation of ADSCs significantly decreased during serial passage. Then, senescence ß-galactosidase (SA-ß-gal) staining showed that the positive rate of P9 ADSCs was 7.74-fold than P3 ADSCs. Subsequently, RNA sequencing (RNA-seq) was performed for P3 and P9 ADSCs and found that PI3K-AKT pathway was up-regulated, but cell cycle and DNA repair pathway were down-regulated in P9 ADSCs. Then, N-Acetylcysteine (NAC) was added during long-term expansion and showed that NAC enhanced the ADSCs proliferation and maintained adipogenic differentiation. Finally, RNA-seq was performed for P9 ADSCs cultured with or without NAC and showed that NAC restored the cell cycle and DNA repair pathway in P9 ADSCs. These results highlighted that NAC was an excellent supplement for large-scale expansion of porcine ADSCs for cultured meat.

Evaluation the food safety of cultured fat via detection of residues of adipogenic differentiation cocktail in cultured fat with high performance liquid chromatography and enzyme-linked immunosorbent assay.

Cultured fat is inducing adipose progenitor cells (APCs) to differentiate into mature adipocytes for consumption. The traditional adipogenic differentiation cocktail, including insulin, dexamethasone, indomethacin, isobutylmethylxanthine and rosiglitazone, has potential food safety problems in cultured fat. Therefore, the detection of these residues is necessary to ensure food safety. In this research, a method of high performance liquid chromatography (HPLC) was established to quantitatively analyze the potential residual content of dexamethasone, indomethacin, isobutylmethylxanthine and rosiglitazone in cultured fat and medium. Quantitative analysis showed that the content of four residues in cultured fat decreased to zero on Day 10. Subsequently, enzyme-linked immunosorbent assay (ELISA) was performed to detect the insulin content in the cultured fat and found that the insulin content in the cultured fat on Day 10 was 2.78 ± 0.21 µg/kg. After soaking with phosphate buffered saline (PBS), the insulin content decreased to 1.88 ± 0.54 µg/kg. In conclusion, this research provided an effective approach to clarify the content of potential residual components in cultured fat and it will provide reference for the safety of cultured fat in the future.

The inhibitory effect of p75 neurotrophin receptor on growth of human hepatocellular carcinoma cells.

p75 neurotrophin receptor (p75NTR), a member of the TNF receptor superfamily, is a focus for study at present. Up to now, its role and functions in hepatocellular carcinoma were not fully elucidated. In this study, we investigated the expression of p75NTR in hepatocellular carcinoma and the impact of its alteration on tumor growth. We found that the expression of p75NTR was decreased significantly in 158 cases of hepatocellular carcinoma tissues as compared with their adjacent noncancerous counterparts, and its expression was also significantly decreased in various human hepatocellular carcinoma cell lines. Down-regulating p75NTR by specific siRNA promoted the growth of normal liver cell lines, whereas up-regulating p75NTR inhibited the growth of hepatocellular carcinoma cell lines in vitro and caused dramatic attenuation of tumor growth in vivo by induction of cell cycle arrest. Furthermore, we found that up-regulating p75NTR could down-regulate the expression of cyclin A, cyclin D1, cyclin E, cdk2, p-Rb and PCNA, but up-regulate the expression of Rb. Conversely, the results were inverse when p75NTR was down-regulated by specific siRNA. Therefore, we provided the evidence that p75NTR was a potential tumor suppressor and might be used as a therapeutic target for hepatocellular carcinoma.

Injectable hydrogel microspheres experimental research for the treatment of osteoarthritis / 中国修复重建外科杂志

OBJECTIVE@#To prepare a novel hyaluronic acid methacrylate (HAMA) hydrogel microspheres loaded polyhedral oligomeric silsesquioxane-diclofenac sodium (POSS-DS) patricles, then investigate its physicochemical characteristics and in vitro and in vivo biological properties.@*METHODS@#Using sulfhydryl POSS (POSS-SH) as a nano-construction platform, polyethylene glycol and DS were chemically linked through the "click chemistry" method to construct functional nanoparticle POSS-DS. The composition was analyzed by nuclear magnetic resonance spectroscopy and the morphology was characterized by transmission electron microscopy. In order to achieve drug sustained release, POSS-DS was encapsulated in HAMA, and hybrid hydrogel microspheres were prepared by microfluidic technology, namely HAMA@POSS-DS. The morphology of the hybrid hydrogel microspheres was characterized by optical microscope and scanning electron microscope. The in vitro degradation and drug release efficiency were observed. Cell counting kit 8 (CCK-8) and live/dead staining were used to detect the effect on chondrocyte proliferation. Moreover, a chondrocyte inflammation model was constructed and cultured with HAMA@POSS-DS. The relevant inflammatory indicators, including collagen type Ⅱ, aggrecan (AGG), matrix metalloproteinase 13 (MMP-13), recombinant A disintegrin and metalloproteinase with thrombospondin 5 (Adamts5), and recombinant tachykinin precursor 1 (TAC1) were detected by immunofluorescence staining and real-time fluorescence quantitative PCR, with normal cultured chondrocytes and the chondrocyte inflammation model without treatment as control group and blank group respectively to further evaluate their anti-inflammatory activity. Finally, by constructing a rat model of knee osteoarthritis, the effectiveness of HAMA@POSS-DS on osteoarthritis was evaluated by X-ray film and Micro-CT examination.@*RESULTS@#The overall particle size of POSS-DS nanoparticles was uniform with a diameter of about 100 nm. HAMA@POSS-DS hydrogel microspheres were opaque spheres with a diameter of about 100 μm and a spherical porous structure. The degradation period was 9 weeks, during which the loaded POSS-DS nanoparticles were slowly released. CCK-8 and live/dead staining showed no obvious cytotoxicity at HAMA@POSS-DS, and POSS-DS released by HAMA@POSS-DS significantly promoted cell proliferation (P<0.05). In the chondrocyte anti-inflammatory experiment, the relative expression of collagen type Ⅱ mRNA in HAMA@POSS-DS group was significantly higher than that in control group and blank group (P<0.05). The relative expression level of AGG mRNA was significantly higher than that of blank group (P<0.05). The relative expressions of MMP-13, Adamts5, and TAC1 mRNA in HAMA@POSS-DS group were significantly lower than those in blank group (P<0.05). In vivo experiments showed that the joint space width decreased after operation in rats with osteoarthritis, but HAMA@POSS-DS delayed the process of joint space narrowing and significantly improved the periarticular osteophytosis (P<0.05).@*CONCLUSION@#HAMA@POSS-DS can effectively regulate the local inflammatory microenvironment and significantly promote chondrocyte proliferation, which is conducive to promoting cartilage regeneration and repair in osteoarthritis.

Application value of SHOX2 and RASSF1A gene promoter region methylation detection for screening and diagnosis of early-stage lung adenocarcinoma / 肿瘤研究与临床

Objective:To investigate the value of SHOX2 and RASSF1A gene promoter region methylation detection for screening and diagnosis of early-stage lung adenocarcinoma.Methods:The mRNA sequencing data of 471 lung adenocarcinoma patients and corresponding methylation data of 413 cases were downloaded from The Cancer Genome Atlas (TCGA) database, the methylation levels of SHOX2 and RASSF1A gene promoter regions were calculated, and the difference in methy lation level between normal lung tissues and tumor tissues was analyzed. The clinical data of 54 patients with early-stage lung adenocarcinoma and 31 patients with benign lung tumors who underwent surgery at Drum Tower Hospital Affiliated to Nanjing University Medical School from January 2018 to January 2019 were retrospectively analyzed. The methylation status of SHOX2 and RASSF1A in tumor tissues and normal lung tissues (>5 cm from the edge of the tumor foci) (called clinical samples) was detect, and a positive methylation in the promoter region of either gene was considered as a combination of two genes methylation positivity. Using pathological diagnosis as the gold standard, the efficacy of gene methylation positivity in diagnosing early-stage lung adenocarcinoma was analyzed by receiver operating characteristic (ROC) curve. Patients with >80% of tumor cells in paraffin samples were screened, and mRNA high-throughput sequencing was performed in their tumor tissues and normal lung tissues. The relationship between positive methylation of the two genes and clinicopathological features was analyzed, and the correlation between the promoter region methylation level of the two genes and mRNA expression levels in clinical samples and TCGA database samples was analyzed by Spearman method. Gene set variance analysis (GSVA) method was used to analyze the differences in Kyoto Encyclopedia of Genes and Genomes enrichment pathways between two-gene methylation-positive clinical lung adenocarcinoma samples and corresponding methylation-negative lung adenocarcinoma.Results:In TCGA database, the SHOX2 promoter region methylation island contained 6 sequenced methylation sites, of which sites cg04532033 and cg01557547 methylation levels were higher in lung adenocarcinoma tissues than in normal lung tissues (both P < 0.05); the RASSF1A gene promoter region methylation island contained 11 sequenced methylation sites, and the methylation levels of 6 of these sites in lung adenocarcinoma tissues were higher than those in normal lung tissues (all P < 0.05). Compared with normal lung tissues, the methylation level of SHOX2 promoter region was higher in stage Ⅰ and Ⅱ lung adenocarcinoma tissues (both P < 0.05); the methylation level of RASSF1A promoter region was higher in all stages of lung adenocarcinoma ( P < 0.001). Among 54 patients with early-stage lung adenocarcinoma, 28 were positive for SHOX2 promoter region methylation in tumor tissues, 21 were positive for RASSF1A promoter region methylation, and 40 were positive for combined methylation of both genes; 31 benign lung nodules were negative for SHOX2 and RASSF1A methylation. ROC curve analysis showed that the sensitivity of positive SHOX2 promoter region methylation for diagnosing early-stage lung adenocarcinoma was higher than that of RASSF1A promoter region methylation positivity (51.8% vs. 38.9%), and the area under the curve (AUC) for diagnosis by two-gene methylation positivity was larger than that for diagnosis by SHOX2 or RASSF1A gene methylation positivity alone (0.870 vs. 0.759 and 0.694). The circulating thresholds (Ct) of SHOX2 and RASSF1A methylation tested by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) in stage Ⅰ and Ⅱ lung adenocarcinoma were lower than those in normal lung tissues (all P < 0.05); patients with two-gene methylation positivity were characterized by older age, longer tumor longest diameter and more advanced pathological stage compared with patients with two-gene methylation negativity (all P < 0.05). In clinical stage Ⅰ-Ⅱ lung adenocarcinoma samples, the Ct of SHOX2 and RASSF1A promoter region methylation tested by qRT-PCR was negatively correlated with their mRNA relative expression levels ( r=-0.43, P = 0.003; r = -0.48, P = 0.001); in TCGA database stage Ⅰ-Ⅱ lung adenocarcinoma samples, the level of SHOX2 promoter region methylation was negatively correlated with its mRNA relative expression level ( r = -0.23, P < 0.001), and the level of RASSF1A promoter region methylation was also negatively correlated with its mRNA relative expression level, but without statistical difference ( r = -0.05, P = 0.310). In two-gene promoter methylation-positive lung adenocarcinoma samples, the pathways related to folate metabolism and DNA stability were upregulated, and the pathways related to vasoconstriction and cell growth and differentiation were downregulated. Conclusions:The combined detection of SHOX2 and RASSF1A promoter region methylation can be used as an indicator for screening and diagnosis of early-stage lung adenocarcinoma. Abnormal promoter region methylation of the two genes may affect multiple tumor-related pathways and promote the occurrence and progression of early-stage lung adenocarcinoma.

Preliminary study on the ability of 68Ga-Pentixafor PET/CT to differentiate between adrenal aldosterone-producing adenoma and nonfunctional adenoma / 中华内科杂志

Objective:To evaluate the ability of 68Ga-Pentixafor (nuclide ligand imaging agents for chemokine receptor 4) PET/CT to differentiate between aldosterone-producing adenoma (APA) and adrenal nonfunctional adenoma (NFA), and to assess how well this imaging method correlates with clinical features and postoperative outcomes. Methods:This was a cross-sectional study involving 73 APA and 12 NFA patients who received 68Ga-Pentixafor PET/CT imaging at Peking Union Medical College Hospital from August 2018 to October 2021. The receiver operating characteristic (ROC) curve was used to evaluate the differential value of visual analysis and the maximum standard uptake value (SUV max) of the focus on APA and NFA. The related factors of SUV max, and its predictive effect on postoperative outcomes were analyzed using Pearson or Spearman analysis and χ2 text. Results:68Ga-Pentixafor PET/CT imaging was positive in 64 APA patients (sensitivity=87.7%) and negative in all 12 NFA patients (specificity=100%). The area under the ROC curve with SUV max differentiating APA and NFA was 0.932 ( P<0.001). When the SUV max cut-off point was 6.23, the sensitivity was 80.8% and the specificity was 100%. The SUV max correlated positively with lesion size ( r=0.598) and aldosterone/renin activity ratio ( r=0.313) and correlated negatively with potassium level ( r=-0.286), renin activity ( r=-0.240) and age of diagnosis ( r=-0.273) (all P<0.05). Of the patients who underwent adrenalectomy and received more than 6 months of post-surgical follow-up, the clinical complete remission rate was higher for 68Ga-Pentixafor PET/CT imaging-positive patients than imaging-negative patients (24/39 vs. 0/4, P=0.031). Conclusions:68Ga-Pentixafor PET/CT is effective at differentiating between APA and NFA. The SUV max of 68Ga-Pentixafor PET/CT correlates with age at onset, lesion size, and the severity of clinical manifestations, and is able to predict postoperative outcomes.

Role of new serological markers in evaluating the progression of liver disease in patients with chronic HBV infection and normal aminotransferases / 临床肝胆病杂志

Antiviral therapy is the basic treatment method for improving prognosis recommended in the management guidelines of chronic hepatitis B in China and globally. For patients with chronic HBV infection and normal transaminases, it is difficult in clinical practice to accurately evaluate the progression of hepatitis and identify suitable patients who need antiviral therapy. In order to objectively and accurately evaluate the degree of liver inflammatory activity in such patients, more and more noninvasive evaluation indicators have been used in addition to conventional liver biopsy. This article reviews the new serological indicators that can reflect the degree of liver inflammation and/or fibrosis in patients with chronic HBV infection and normal aminotransferase levels, hoping to provide a reference for antiviral decision-making in these patients.

Specimen extraction through natural orifices with Cai tubes in gastrointestinal surgery: a single-institute series of 234 cases / 中华胃肠外科杂志

Objective: To investigate the feasibility of Cai tube-assisted natural orifice specimen extraction surgery (NOSES) in gastrointestinal surgery. Methods: This was a descriptive case-series study. Inclusion criteria: (1) colorectal or gastric cancer diagnosed by preoperative pathological examination or redundant sigmoid or transverse colon detected by barium enema; (2) indications for laparoscopic surgery; (3) body mass index <30 kg/m2 (transanal surgery) and 35 kg/m2 (transvaginal surgery); (4) no vaginal stenosis or adhesions in female patients undergoing transvaginal specimen extraction; and (5) patients with redundant colon aged 18-70 years and a history of intractable constipation for more than 10 years. Exclusion criteria: (1) colorectal cancer with intestinal perforation or obstruction, or gastric cancer with gastric perforation, gastric hemorrhage, or pyloric obstruction; (2) simultaneous resection of lung, bone, or liver metastases ; (3) history of major abdominal surgery or intestinal adhesions; and (4) incomplete clinical data. From January 2014 to October 2022, 209 patients with gastrointestinal tumors and 25 with redundant colons who met the above criteria were treated by NOSES utilizing a Cai tube (China invention patent number:ZL201410168748.2) in the Department of Gastrointestinal Surgery, Zhongshan Hospital, Xiamen University. The procedures included eversion and pull-out NOSES radical resection in 14 patients with middle and low rectal cancer, NOSES radical left hemicolectomy in 171 patients with left-sided colorectal cancer, NOSES radical right hemicolectomy in 12 patients with right-sided colon cancer, NOSES systematic mesogastric resection in 12 patients with gastric cancer, and NOSES subtotal colectomy in 25 patients with redundant colons. All specimens were collected by using an in-house-made anal cannula (Cai tube) with no auxiliary incisions. The primary outcomes included 1-year recurrence-free survival (RFS) and postoperative complications. Results: Among 234 patients, 116 were male and 118 were female. The mean age was (56.6±10.9) years. NOSES was successfully completed in all patients without conversion to open surgery or procedure-related death. The negative rate of circumferential resection margin was 98.8% (169/171) with both two positive cases having left-sided colorectal cancer. Postoperative complications occurred in 37 patients (15.8%), including 11 cases (4.7%) of anastomotic leakage, 3 cases(1.3%) of anastomotic bleeding, 2 cases (0.9%) of intraperitoneal bleeding, 4 cases (1.7%) of abdominal infection, and 8 cases (3.4%) of pulmonary infection. Reoperations were required in 7 patients (3.0%), all of whom consented to creation of an ileostomy after anastomotic leakage. The total readmission rate within 30 days after surgery was 0.9% (2/234). After a follow-up of (18.3±3.6) months, the 1-year RFS was 94.7%. Five of 209 patients (2.4%) with gastrointestinal tumors had local recurrence, all of which was anastomotic recurrence. Sixteen patients (7.7%) developed distant metastases, including liver metastases(n=8), lung metastases(n=6), and bone metastases (n=2). Conclusion: NOSES assisted by Cai tube is feasible and safe in radical resection of gastrointestinal tumors and subtotal colectomy for redundant colon.

High-fat intake alleviates lung injury induced by Paragonimus proliferus infection in rats through up-regulating CYP 4A1 expression in lung tissues / 中国血吸虫病防治杂志

Objective To explore the improvements of high-fat intake on lung injury induced by Paragonimus proliferus infection in rats, and to preliminarily explore the mechanisms underlying the role of cytochrome P450 4A1 (CYP 4A1) in the improve ments. Methods SD rats were randomly assigned into three groups, including the normal control group (n = 10), the infection and normal diet group (n = 12) and the infection and high-fat diet group (n = 12). Rats in the normal control group were fed with normal diet and without any other treatments, and animals in the infection and normal diet group were subcutaneously injected with 8 excysted metacercariae of P. proliferus via the abdominal wall, followed by feeding with normal diet, while rats in the infection and high-fat diet group were subcutaneously injected with 8 excysted metacercariae of P. proliferus via the abdominal wall, followed by feeding with high-fat diet. All rats were sacrificed 28 weeks post-infection, and serum samples and lung specimens were collected. Following hematoxylin-eosin (HE) staining of rat lung specimens, the rat lung injury was observed under an optical microscope, and alveolitis was evaluated using semi-quantitative scoring. Serum interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) levels were measured using enzyme-linked immunosorbent assay (ELISA), and the cytochrome P450 4A1 (CYP 4A1) expression was quantified in rat lung specimens at transcriptional and translational levels using quantitative real-time PCR (qPCR) and Western blotting assays. Results Alveolar wall thickening, edema and inflammatory cell infiltration were alleviated 28 weeks post-infection with P. proliferus in rats in the infection and high-fat diet group relative to the infection and normal diet group, and no alveolar consolidation was seen in the infection and high-fat diet group. The semi-quantitative score of alveolitis was significantly higher in the infection and normal diet group [(2.200 ± 0.289) points] than in the normal control group [(0.300 ± 0.083) points] and the infection and high-fat diet group [(1.300 ± 0.475) points] (both P values < 0.05), and higher serum IL-1β [(151.586 ± 20.492)] pg/mL and TNF-α levels [(180.207 ± 23.379) pg/mL] were detected in the infection and normal diet group than in the normal control group [IL-1β: (103.226 ± 3.366) pg/mL; TNF-α: (144.807 ± 1.348) pg/mL] and the infection and high-fat diet group [IL-1β: (110.131 ± 12.946) pg/mL; TNF-α: (131.764 ± 27.831) pg/mL] (all P values < 0.05). In addition, lower CYP 4A1 mRNA (3.00 ± 0.81) and protein expression (0.40 ± 0.02) was quantified in lung specimens in the infection and normal diet group than in the normal control group [(5.03 ± 2.05) and (0.84 ± 0.14)] and the infection and high-fat diet group [(11.19 ± 3.51) and (0.68 ± 0.18)] (all P values < 0.05). Conclusion High-fat intake may alleviate lung injuries caused by P. proliferus infection in rats through up-regulating CYP 4A1 expression in lung tissues at both translational and transcriptional levels.

Expression and significance of TWIST basic helix-loop-helix protein over-expression in gastric cancer.

AIMS: TWIST protein has been implicated in neoplastic transformation and development of some cancers. In this study, we aimed to investigate the expression of TWIST in gastric cancer and its clinical significance. METHODS: A total of 76 cases of archival gastric cancer tissues were immunohistochemically evaluated for TWIST expression, and its expression was correlated with clinicopathological parameters. Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the mRNA of TWIST in four gastric cancer cell lines and a normal immortalised gastric epithelial cell line (GES-1). The expression of TWIST protein in these cell lines and 14 pairs of fresh gastric carcinoma and adjacent normal tissue samples was detected by Western blotting. RESULTS: TWIST expression increased in diffuse-type gastric carcinoma compared with intestinal-type gastric carcinoma (26/42, 61.9% versus 9/34, 26.5%, p<0.05). TWIST expression was significantly increased in 35 (46.1%) of the 76 cancers and correlated with lymph node metastasis (node positive rate 60.4%; node negative rate 21.4%; p<0.05). The expression of TWIST protein was higher in 9/14 (64.3%) fresh cancer tissues compared with adjacent normal tissues. The expression of mRNA and protein of TWIST in gastric cancer cell lines was up-regulated compared with that in GES-1. CONCLUSIONS: TWIST was highly expressed in gastric cancer. Its up-regulation was associated with the neoplastic transformation and subsequent development of gastric cancer. Therefore, TWIST may be a useful prognostic marker and target for gastric cancer therapy.

Efficacy of concentrated growth factor in the reduction of complications after bone augmentation during surgery for odontogenic maxillary cyst / 中国基层医药

Objective:To investigate the efficacy of concentrated growth factor (CGF) in the reduction of pain and swelling after bone augmentation during surgery for odontogenic maxillary cyst.Methods:Sixteen patients with odontogenic maxillary cysts who underwent bone augmentation during surgery between June 2018 and December 2019 were included in this study. They were randomly divided into two groups ( n = 8/group). In the observation group, CGF and bone substitutes were mixed and covered with biofilm. In the control group, bone powder was used to fill the bone cavity, and artificial biofilm was applied. Postoperative swelling, pain, and scar hyperplasia were compared between the two groups. Pain was rated according to visual analogue score. Swelling was divided into three grades according to the level of swelling. Scar was compared according to vancouver scar scale. Statistical analysis was performed using SPSS 20.0 software. Results:The percentage of grade 1-3 swelling in the observation group were 62.5% (5/8), 37.5% (3/8) and 0.0% (0/8), respectively, and they were 50.0% (4/8), 37.5% (3/8), 12.5%(1/8), respectively ( Z = -1.71, P > 0.05). There was significant difference in the duration of swelling between observation and control groups [(3.8 ± 0.9) days vs. (5.8 ± 1.4) days, t = 6.88, P < 0.05]. At 3, 7 and 14 days after surgery, visual analogue score was (2.21 ± 0.25) points, (3.75 ± 0.22) points and (0.57 ± 0.13) points in the observation group and it was (3.76 ± 0.18) points, (2.38 ± 0.26) points, and (2.38 ± 0.26) points in the control group ( t = 14.23, t = 11.29, t = 17.61, all P < 0.001). At 1, 2 and 4 weeks after surgery, vancouver scar scale score was (4.26 ± 0.26) points, (1.22 ± 0.13) points and (2.47 ± 0.11) points in the observation group, and it was (6.35 ± 0.27) points, (4.47 ± 0.73) points and (2.77 ± 0.21) points in the control group ( t = 15.77, t = 2.67, t = 3.58, P < 0.001). Conclusion:CGF can promote postoperative bone defect repair and wound healing and reduce postoperative swelling, pain and scar.

The Status Quo and Prospects of Diagnosis and Treatment of Rare Kidney Disease in Children / 罕见病研究

Rare kidney diseases are important causes of chronic kidney disease (CKD) in children. The majority of rare kidney diseases in children are hereditary kidney diseases, accounting for about 29.7% to 52.1% of children with CKD. Next-generation sequencing has been widely used in clinical diagnosis in the past decade, leading to the improvement of the diagnosis of hereditary kidney diseases. In 2018, China announced the first list of rare diseases and greatly enhanced the diagnosis, treatment and research of rare diseases in China, including rare kidney disease. Meanwhile, China faces great challenges in the diagnosis and treatment of hereditary kidney diseases in Children, including the assessment of pathogenicity of gene variants, the lack of biomarkers for disease progression and therapy efficacy, lack of drugs, and others. The future lies in the cooperation between patients, physicians, researchers, and health policy markers, and the fast translation from research finding to clinical application, so as to meet the demand from the children with rare kidney diseases in China.

Imaging evaluation of vulnerability of intracranial atherosclerotic plaques / 国际脑血管病杂志

Intracranial atherosclerotic disease (ICAD) is an important cause of ischemic stroke. Accurate clinical and imaging evaluation is helpful to its hierarchical management and individualized treatment. With the gradual maturation of intracranial artery wall imaging technology, the stroke mechanism of ICAD can be further understood through plaque vulnerability characteristics and the optimal stroke prevention strategy can be developed. This article reviews the different vulnerable characteristics, evaluation methods and current research progress of intracranial atherosclerotic plaques.

Analysis of immune microenvironment and immune-related differentially expressed genes related to prognosis in metastatic colorectal cancer / 肿瘤研究与临床

Objective:To analyze the difference in immune microenvironment between primary tumor tissues and metastatic tumor tissues of metastatic colorectal cancer, and to screen specific immune-related differentially expressed genes (DEG) related to prognosis of metastatic colorectal cancer via bioinformatics methods.Methods:The GSE131418 microarray dataset of colorectal cancer and metastases was downloaded from gene expression omnibus (GEO) database, including 517 samples from the MCC cohort and 618 samples from the Consortium cohort in Moffitt Cancer Center. Immune-related gene sets were downloaded from immunology database and analysis portal IMMPORT, including 2 483 immune-related genes. A total of 695 cases of RNA sequencing data and 627 cases of clinical information of colorectal cancer tumors and adjacent tissues were downloaded from Cancer Genome Atlas (TCGA) data. The stroma cell score, immune cell score and stromal immune total score of metastatic tumor tissues and primary tumor tissues were calculated by using ESTIMATE algorithm, and 22 kinds of immune cell infiltration in primary tumor and metastatic tumor tissues of colorectal cancer were compared and analyzed by using CIBERSORT deconvolution algorithm. Immune-related DEG were screened to make Kyoto Encyclopedia of Genes and Gnomes (KEGG) signaling pathway enrichment analysis. The patients were divided into high and low expression groups according to the median expression levels of immune-related DEG. The Kaplan-Meier method and Cox regression risk model were used to analyze immune-related DEG, and the genes significantly related to prognosis in the results of the two methods were screened (all P < 0.01), and multivariate analysis was performed by using Cox regression method. The expression differences of each gene in tumor tissues, adjacent tissues, primary tumor tissues and metastatic tissues in GSE131418 data sets of TCGA database and GEO database were compared, and survival analysis was also performed. Results:The stroma cell score, immune cell score and stromal immune total score of colorectal cancer metastatic tissues were lower than those of primary tumor tissues (all P < 0.001). Compared with primary tumor tissues, the proportion of activated natural killer (NK) cells, monocytes, CD8 + T cells, T cells, activated dendritic cells in metastatic colorectal cancer tissues was increased, while the proportion of inactive mast cells, inactive dendritic cells, inactive NK cells, activated memory CD4 + T cells, M1 macrophages, and neutrophils was decreased. There were 289 immune-related DEG in metastatic tissues and primary tumor tissues of metastatic colorectal cancer, including 101 up-regulated genes and 188 down-regulated genes. KEGG signaling pathway enrichment analysis showed that in the immune microenvironment of metastatic tissues in metastatic colorectal cancer, vascular endothelial growth factor (VEGF) signaling pathway, programmed death ligand 1 (PD-L1) expression and programmed death 1 (PD-1) checkpoint pathway, T helper cell (Th) 1, Th2 and Th17 cell differentiation, NF-kappa B signaling pathway, interleukin 17 (IL-17) signaling pathway, chemokine signaling pathway, T cell receptor signaling pathway, MAPK signaling pathway, and NK cell-mediated cytotoxicity pathways enrichment were detected. Immune-related DEG related to prognosis including ANGPTL5, FPR1, HSPA8, NR2E3, PSMD2, PSMD8 and SBDS were screened out. Cox regression multivariate analysis showed that immune-related DEG ANGPTL5 ( HR = 2.69, 95% CI 1.22-5.92, P < 0.05), HSPA8 ( HR = 0.57, 95% CI 0.33-0.97, P < 0.05), and SBDS ( HR = 2.23, 95% CI 1.18-4.21, P < 0.05) were independent prognostic factors for metastatic colorectal cancer. The expression of ANGPTL5 in tumor tissues was lower than that in normal tissues, and the expression of ANGPTL5 in metastatic tissues was higher than that in primary tumor tissues. Patients with high expression of ANGPTL5 in tumor tissues had worse prognosis. The expression of HSPA8 in tumor tissues was higher than that in normal tissues, and the expression of HSPA8 in metastatic tissues was lower than that in primary tumor tissues. Patients with high expression of HSPA8 in tumor tissues had a better prognosis. The expression of SBDS in tumor tissues was lower than that in normal tissues, and the expression of SBDS in metastatic tissues was lower than that in primary tumor tissues. Patients with high expression of SBDS in tumor tissues had worse prognosis. Conclusions:Immune microenvironment of metastatic colorectal cancer is quite different from that of primary tumor. The degree of immune cell infiltration is reduced and the whole is immunosuppressed. The specific immune-related DEG related to prognosis of metastatic colorectal cancer may be new therapeutic targets of metastatic colorectal cancer.