A recurrence-predictive model based on eight genes and tumor mutational burden/microsatellite instability status in Stage II/III colorectal cancer.

BACKGROUND: Although adjuvant chemotherapy (ACT) is widely used to treat patients with Stage II/III colorectal cancer (CRC), administering ACT to specific patients remains a challenge. The decision to ACT requires an accurate assessment of recurrence risk and absolute treatment benefit. However, the traditional TNM staging system does not accurately assess a patient's individual risk of recurrence. METHODS: To identify recurrence risk-related genetic factors for Stage II/III CRC patients after radical surgery, we conducted an analysis of whole-exome sequencing of 47 patients with Stage II/III CRC who underwent radical surgery at five institutions. Patients were grouped into non-recurrence group (NR, n = 24, recurrence-free survival [RFS] > 5 years) and recurrence group (R, n = 23, RFS <2 years). The TCGA-COAD/READ cohort was employed as the validation dataset. RESULTS: A recurrence-predictive model (G8plus score) based on eight gene (CUL9, PCDHA12, HECTD3, DCX, SMARCA2, FAM193A, AATK, and SORCS2) mutations and tumor mutation burden/microsatellite instability (TMB/MSI) status was constructed, with 97.87% accuracy in our data and 100% negative predictive value in the TCGA-COAD/READ cohort. For the TCGA-COAD/READ cohort, the G8plus-high group had better RFS (HR = 0.22, p = 0.024); the G8plus-high tumors had significantly more infiltrated immune cell types, higher tertiary lymphoid structure signature scores, and higher immunological signature scores. The G8plus score was also a predict biomarker for immunotherapeutic in advanced CRC in the PUCH cohort. CONCLUSIONS: In conclusion, the G8plus score is a powerful biomarker for predicting the risk of recurrence in patients with stage II/III CRC. It can be used to stratify patients who benefit from ACT and immunotherapy.

Analysis of the decoupling effect and driving factors of carbon emissions from the transportation sector in Guangdong Province.

To propel the green transformation of the transportation industry in Guangdong Province. Against the backdrop of the five-year plan, this study employs the Tapio decoupling model and the Logarithmic Mean Divisia Index decomposition method to analyze the decoupling status and driving factors of carbon emissions from transportation and economic growth in Guangdong Province from 2001 to 2020. The results show that:(1) Both the total volume of carbon emissions from transportation and per capita emissions exhibit an overall upward trend, with petroleum-based emissions accounting for an overwhelming 96%, with diesel emissions register the highest proportion and most substantial increase. (2) The decoupling status predominantly manifests as weak decoupling, with intermittent years expansive coupling,, expansive negative decoupling, strong decoupling, and recessive coupling, thus indicating a persistent state of decoupling instability. (3) The study introduces the urbanization effect, wherein the effects of income urbanization, as well as spatial urbanization, predominantly exert positive driving influences. Conversely, the effects of energy intensity, population urbanization, industry scale, and energy structure collectively exert negative driving influences. Notably, the effect of income urbanization emerges as the primary factor propelling the augmentation of carbon emissions, while the energy intensity effect serves as the primary factor curbing such escalation. Finally, pertinent policy recommendations are put forth.

Aberrant expression of WNK lysine deficient protein kinase 1 is associated with poor prognosis of colon adenocarcinoma.

BACKGROUNDS: WNK1 (WNK lysine deficient protein kinase 1) is a kind of protein kinase and participates in angiogenesis, having a potent tumor promoting role. WNK1 is ubiquitously expressed, and its upregulated expression has been reported in several tumor types. AIMS: Here, we aimed to investigate the correlation between WNK1 expression and colon adenocarcinoma (COAD) progression. METHODS: In the current study, WNK1 expression was evaluated by immunohistochemically in adjacent normal colonic mucosae and primary adenocarcinomas. The effect of WNK1 on overall survival (OS) and its associations with the clinicopathological parameters were analyzed in a retrospective cohort of COAD patients (n = 185). The tumor-related effects of WNK1 in COAD were further tested via cellular and mice experiments. RESULTS: According to our cohort, higher WNK1 expression was significantly associated with unfavorable prognostic factors, such as high pT stage, pN stage, as well as shorter OS. Moreover, WNK1 exhibited tumor promoting role in COAD cancer cell lines as well as in nude mice. Silencing WNK1 can significantly inhibit the proliferation of COAD both in vitro and in vivo. CONCLUSIONS: In all, WNK1 acts as a tumor promoter and may be used as a COAD prognostic biomarker.

5-azacytidine inhibits Sox2 promoter methylation during the induction of Thy-1+Lin- cells into hepatocytes.

OBJECTIVE: To investigate the changes and functions of Sox2 gene expression and promoter methylation during induced differentiation of bone marrow mesenchymal stem cells (BMSCs) into hepatocytes (HCs). METHODS: Rat bone marrow Thy-1+Lin- cells were prepared and divided into control group (directed induction of differentiation into HCs) and experimental group (5-azacytidine intervention induced differentiation). The mRNA expression levels of ALB and Sox2 were detected by fluorescence quantitative polymerase chain reaction (PCR), and the Sox2 gene promoter methylation level was determined by Bisulfite sequencing PCR (BSP). RESULTS: Sox mRNA expression level was significantly increased in experimental group compared to the control group at 0, 7, and 14 days, respectively (all P<0.05). The Sox2 promoter methylation level was gradually increased after 0, 7 and 14 days induction in both groups, accompanied by an increase in methylated loci (all P<0.05). Statistical significance was present in CpG methylated loci between groups (all P<0.05). CONCLUSIONS: The expression of Sox2 gene increased first and then decreased in the process of inducing rat BMSCs into stem cells, and the methylation level of CpG loci in the promoter region changed dynamically, with an increased overall methylation level. After 5-aza treatment, the Sox2 promoter was in a non-methylated state, and its mRNA expression increased, which hindered the cell differentiation.

Effect of aeration on water quality and sediment humus in rural black-odorous water.

Black-odorous water, which exists widely in rural areas of China, affects the living environments of residents and causes a loss of ecosystem functions, and improper treatment may even cause secondary pollution. Moreover, the transformation relationships among the components of humus in the sediments of black-odorous water are largely unknown. Therefore, it is necessary to select appropriate methods for treating black-odorous water in rural areas and to understand the characteristics of humus in sediment during the treatment process. In this study, the use of aeration in treating black-odorous water and interstitial water in rural areas was studied and the effects of different treatment methods on the contents and spectral characteristics of humus components in sediments were explored. It was found that the quality of the treated water improved from severe black-odorous to nonblack-odorous, the colour of the sediment changed from black to greyish-brown to turquoise, and the odour changed from strong to weak. The removal rates of ammonia nitrogen, total nitrogen, total phosphorus, and chemical oxygen demand in the sediment aeration group reached more than 50% in the upper water, and more than 70% in the interstitial water. After treatment, the content of fulvic acid, the main black substance in sediment, decreased by 0.36-1.58 g/kg, and the molecular structure of the humus was simplified.

Methodology of emergency medical logistics for multiple epidemic areas in public health emergency.

Coronavirus disease 2019(COVID-19) has brought great disasters to humanity, and its influence continues to intensify. In response to the public health emergencies, prompt relief supplies are key to reduce the damage. This paper presents a method of emergency medical logistics to quick response to emergency epidemics. The methodology includes two recursive mechanisms: (1) the time-varying forecasting of medical relief demand according to a modified susceptible-exposed-infected- Asymptomatic- recovered (SEIAR) epidemic diffusion model, (2) the relief supplies distribution based on a multi-objective dynamic stochastic programming model. Specially, the distribution model addresses a hypothetical network of emergency medical logistics with considering emergency medical reserve centers (EMRCs), epidemic areas and e-commerce warehousing centers as the rescue points. Numerical studies are conducted. The results show that with the cooperation of different epidemic areas and e-commerce warehousing centers, the total cost is 6% lower than without considering cooperation of different epidemic areas, and 9.7% lower than without considering cooperation of e-commerce warehousing centers. Particularly, the total cost is 20% lower than without considering any cooperation. This study demonstrates the importance of cooperation in epidemic prevention, and provides the government with a new idea of emergency relief supplies dispatching, that the rescue efficiency can be improved by mutual rescue between epidemic areas in public health emergency.

Long non-coding RNA FALEC promotes colorectal cancer progression via regulating miR-2116-3p-targeted PIWIL1.

BACKGROUND: Colorectal cancer (CRC) is one of the most common digestive malignant tumors globally. Focally amplified lncRNA on chromosome 1 (FALEC) is a novel lncRNA that has been reported to be involved in many biological processes during carcinogenesis. However, its role in CRC remains poorly understood. METHODS: Gene expression at mRNA or protein level was measured by qRT-PCR or western blot, respectively. In vitro experiments including EdU, colony formation, flow cytometry, wound-healing and transwell assays, as well as in vivo xenograft experiment, were utilized to determine the functional role of FALEC in CRC. Relevant mechanical assays were performed to investigate the underlying molecular mechanism. RESULTS: FALEC was aberrantly up-regulated in CRC. FALEC knockdown could impair CRC cell proliferation, migration and invasion, whereas facilitate cell apoptosis. MiR-2116-3p was revealed to be sponged by FALEC. PIWIL1 was identified as the target of miR-2116-3p. Mechanically, FALEC restored the expression of PIWIL1 via absorbing miR-2116-3p. MiR-2116-3p inhibition and PIWIL1 enrichment could counteract the anti-tumor impact induced by silenced FALEC on the oncogenic behaviors of CRC cells. CONCLUSION: Our study revealed that FALEC promoted CRC progression via restoring the expression of miR-2116-3p-targeted PIWIL1, suggesting the potential application of targeting FALEC in the treatment of CRC.

Novel Presentation of Rosai-Dorfman Histiocytosis With a Prolonged Course of Cranial and Peripheral Neuropathies.

BACKGROUND: Rosai-Dorfman disease is a form of histiocytosis affecting the systemic lymph nodes. Intracranial Rosai-Dorfman disease is rare and presents with extra-parenchymal or intraparenchymal proliferative mass lesions. Cranial neuropathy has not been reported in Rosai-Dorfman disease except when caused by mass effect by an adjacent lesion. PATIENT DESCRIPTION: We describe a girl with Rosai-Dorfman disease who presented with peripheral and multiple cranial neuropathies. Detailed clinical, immunologic, neurophysiology, imaging, and genetic studies were performed. She had a prolonged course but recovered fully after immune therapies. She had increased titers of striated muscle and smooth muscle antibodies. Imaging studies revealed contrast enhancement of cranial nerves and striated muscles. Demyelination was evident in the nerve twigs from muscle biopsy. Exome sequencing did not reveal a genetic mutation. CONCLUSIONS: Most patients with Rosai-Dorfman disease have a benign course, but severe neurological dysfunction due to bulbar involvement and cranial and peripheral neuropathies may occur. Treatment with immunoglobulin and steroids may be of benefit.

Beneficial effect of tetrandrine on refractory epilepsy via suppressing P-glycoprotein.

Patients with refractory epilepsy are resistance to antiepileptic drugs (AEDs). The mechanisms of drug resistance are varied, but one of them is the overexpression of multidrug transporters, such as P-glycoprotein (P-gp), in the brain. Tetrandrine (TTD) is a bis-benzylisoquinoline alkaloid isolated from the root of Stephania tetrandra (S, Moore) and is found to have a favorable effect against multidrug resistance (MDR) in chemotherapy. However, whether TTD affects AEDs in refractory epilepsy is unknown. In this study, we investigated the change in AED treatment efficacy in doxorubicin-induced drug resistant cells after TTD administration. We also examined the effect of TTD on seizure behaviors in the refractory epileptic rats, specifically the expression of MDR1 mRNA and P-gp protein in the cortex and hippocampus of the refractory epileptic rats. Our results demonstrated that TTD decreased cell resistance to phenytoin and valproate. TTD decreased seizure rate and increased the treatment efficacy of AEDs by reducing the expression of P-gp at mRNA and protein levels in vivo. These data support the use of TTD as an adjuvant drug for treating refractory epilepsy.

[Accuracy of endorectal ultrasonography in preoperative staging of rectal cancer].

OBJECTIVE: To evaluate the accuracy of endorectal ultrasonography in preoperative staging of rectal carcinoma. METHODS: The 319 patients with rectal adenocarcinoma underwent endorectal ultrasonography evaluation from January 2007 to March 2010. There were 175 males and 144 females, and the age of patients were 22-82 year old (median 59 years). According their visiting time, 319 patients were divided into 3 groups (period A: January to December 2007; period B: January to December 2008; and period C: January 2009 to March 2010). All patients underwent endorectal ultrasonography, and the 3 doctors had finished evaluations with 272 cases (Doctor 1, 2, 3 had finished evaluations with 162, 64 and 46 cases respectively). The endorectal ultrasonography staging was compared with the pathology findings based on the surgical specimens in 319 patients who had surgery. RESULTS: Overall accuracy in assessing the level of rectal wall invasion was 67%. The accuracy of uT2 and uT3 were 43% and 81% respectively, and the difference was statistically significant (χ(2) = 30.54, P < 0.01), and the accuracy of uT4a was 59%, which was lower than uT3 (81%,χ(2) = 13.77, P < 0.01). Overall accuracy in assessing nodal involvement in the 311 patients treated with radical surgery was 66%. Staging accuracy tends to improve with experience, the accuracy with Doctor 1 in period C(staging accuracy of T and N were 84% and 81% respectively) were higher than period A(staging accuracy of T and N were 55% and 41% respectively) (χ(2) = 6.65 and 13.27, P < 0.01). CONCLUSIONS: Transrectal ultrasound for preoperative staging of rectal has higher accuracy with mastered ultrasound doctor.

[Relationship of c-kit and platelet-derived growth factor receptor alpha gene mutation features with prognosis of patients with gastrointestinal stromal tumor].

OBJECTIVE: To explore the relationship between c-kit and platelet-derived growth factor receptor alpha(PDGFRA) gene mutation features and the prognosis of gastrointestinal stromal tumor(GIST). METHODS: Clinicopathological, genetic testing and follow-up informations of patients admitted to the Shanxi Tumor Hospital from June 2000 to January 2009 were collected. The survival was calculated and univariate analysis was conducted using the Kaplan-Meier method. Multivariate analysis was conducted by the Cox regression method. RESULTS: The 5-year disease-free survival rate was 61.5% and the 5-year overall survival rate was 67.4%. The 5-year disease-free survival rates of patients without disease among those with c-kit exon 11 mutation (n=77), c-kit exon 9 mutation(n=4), and PDGFRA exon 18 mutation (n=2) were 63.4%, 14.3% and 100%, and the 5-year overall survival rates were 70.8%, 50.0% and 100%, respectively. In the patients with c-kit exon 11 mutation, the 5-year disease-free survival rates among those with point mutations(n=26), deletion mutations(n=44), and duplication mutations(n=7) were 87.1%, 44.9% and 80.0%, and the 5-year overall survival rates were 88.1%, 57.0% and 100%, respectively. There were significant differences in overall survival among different factors. Multivariate analysis showed that gene mutation was not the independent factor of prognosis(P=0.492). CONCLUSIONS: In GIST patients undergoing surgery without imatinib treatment, mutated genotype is better than wild type in terms of prognosis. Gene mutation is not the independent factor of prognosis in GIST patients.

[Pathological changes of rectal cancer after irinotecan, 5-fluorouracil or combined short-term radiotherapy].

OBJECTIVE: To observe and evaluate the pathologic changes and curative effects of irinotecan (CPT-11), 5-fluorouracil (5-FU) and combined short-term radiotherapy before low-set rectal cancer operation so as to provide a theoretic basis for formulating a new effective adjuvant therapeutic regimen. METHODS: A total of 41 patients of low rectal cancer were treated with CPT-11, 5-FU therapy or CPT-11 plus 5-FU combined short-term radiotherapy from April 2002 to April 2009. They were divided into 2 groups according to different treatment schemes, including irinotecan group (n = 18) and irinotecan combined short-term radiotherapy group (n = 23). The pathologic changes before and after treatment were observed and the differences of two treatment approaches compared. RESULTS: Tumor cells had different degrees of degeneration and necrosis under microscope in two groups. Compared with computed tomographic findings before therapy, tumor sizes of two groups were reduced by an average of 33.1% (13.5 mm vs 20.2 mm) and 34.4% (12.8 mm vs 19.5 mm) respectively. Two groups were graded according to the RCRG (rectal cancer regression grade) score: RCRG1: 7 cases vs 18 cases, RCRG2: 4 cases vs 3 cases and RCRG3: 7 cases vs 2 cases. According to the pathologic evaluation standard, 3-degree necrosis, cell interstitial fibrosis and intimal thickening in vessels were observed in two groups: 7 cases vs 17 cases, 6 cases vs 17 cases and 3 cases vs 14 cases respectively (all P < 0.05). Five patients achieved complete pathological remission in the irinotecan combined short-term radiotherapy group. CONCLUSION: Based on the pathological changes and mitigation results after treatment, CPT-11 and 5-FU may be used as neoadjuvant drugs for rectal cancer. If the above two drugs can be used in combination with short-term radiation, the curative effect will be better.

[sRNA (sraB) regulate the resistant ability of Salmonella enterica serovar enteritidis to egg albumen].

OBJECTIVE: Salmonella species are important food-borne pathogens of human and animal. S. enterica serovar Enteritidis is the only serovar that routinely causes human infection through intact egg, the molecular basis of its ability to survive in egg is poorly understood. The importance of post-transcriptional regulation by small non-coding RNAs (sRNA) has recently been recognized. The sRNAs play diverse physiological roles in stress responses, regulation of metabolism, control of bacterial envelope composition and bacterial virulence. In this study, we studied regulatory function of salmonella sRNA sraB associated with survival ability of S. enterica serovar Enteritidis in egg albumen. METHODS: To study the contribution of sraB to the survival ability of S. Enteritidis in egg albumen, we constructed sraB deletion strain (SE2472 delta sraB) with wild type S. Enteritidis SE2472, using red recombination system. For complementation of sraB, complete fragment sraB was amplified from SE2472 and inserted into plasmid pHDB3 to overexpress sraB. We carried out the egg albumen bactericidal experiment with strains of SE2472, SE2472 delta sraB (sraB deletion), SE2472 delta sraB-comp (sraB complement) and control. To explore the regulatory role of sraB, we assayed the bactericidal activity of the two important antimicrobial components of egg albumen: lysozyme and transferrin. RESULTS: In the egg albumen bactericidal experiment, the survival rate of SE2472 delta sraB was only about 61%-70% of that of SE2472; while SE2472 delta sraB-comp improve the survival rate of SE2472 delta sraB by 10%-33% . In the transferrin bactericidal experiment, the survival rate of SE2472 delta sraB was 38% of that of SE2472 at 8 h incubation, and 23% at 24 h incubation. SE2472 delta sraB-comp played an important role in improving the survival rate rescued the defect by 14% than SE2472 delta sraB at 8 h of incubation, but failed to rescue the defect at 24 h incubation. In the Lysozyme experiment, the survival rate of SE2472 delta sraB was 41% of that of SE2472 at 8 h incubation, and 27% at 24 h incubation, compared with SE2472 delta sraB, the expression of sraB of SE2472 delta sraB-comp has improved the survival rate by 35% after 8 h of incubation and 23% after 24 h of incubation. CONCLUSION: Based on our results, we conclude that small RNA (sraB) plays important role during the survival of S. Enteritidis in egg albumen, and may contribute regulatory role in response to antimicrobial components of egg albumen such as lysozyme and transferrin.

Minimal role for activating transcription factor 3 in the oligodendrocyte unfolded protein response in vivo.

To further our goal of identifying and characterizing the functions of major components of the unfolded protein response (UPR) in oligodendrocytes, the gene encoding the activator of transcription factor 3 protein (ATF3) has been ablated in mice expressing mutant forms of the Proteolipid protein 1 (Plp1) gene and the phenotype of double mutants characterized at several levels. Mature oligodendrocytes in Plp1 mutant mice undergo UPR-induced cell stress, induce ATF3 expression and exhibit a greater propensity to die by apoptosis, which is consistent with pro-death function of ATF3 proposed from in vitro studies. However, we find that the absence of ATF3 has no effect on the levels of apoptosis in Plp1 mutants. Furthermore, we find that oligodendrocyte function appears normal in Atf3(-/-) mice and that motor coordination and neural communication are similarly unaffected. Accordingly, we conclude that ATF3, at best, plays a minor role in UPR signaling and its expression is more likely induced by the UPR as a secondary event in oligodendrocytes that is unrelated to cell death.

Fatty acid synthase expression during peripheral nervous system myelination.

The expression of fatty acid synthase (FAS) in rat and mouse sciatic nerves during postnatal development was investigated. FAS activity was not sensitive to the nutritional status of the animals. During development, the specific activity of FAS was low in rat and mouse nerves immediately after birth. Then, there was a steady increase in the activity (8- to 10-fold) which reached a maximal level around postnatal day 11, plateaued till day 32, and decreased to reach 30% of the maximum at day 80. A similar developmental profile was obtained when the amount of FAS protein was quantified, thus suggesting that the variations in activity observed during sciatic nerve development are mainly due to variations in FAS protein content. Northern blot analysis showed that the mRNA levels for FAS parallels those of the ceramide galactosyl transferase (CGT) during mouse sciatic nerve development and in a rat demyelination-nerve regeneration model. In addition, we measured FAS expression in the sciatic nerves of the trembler mutant, which is a mouse model of PNS dysmyelination. In 20-day-old trembler nerves, FAS specific activity, protein amount and mRNA levels represented only 25% of the normal values. Altogether, our data indicate that FAS expression is linked to the PNS myelination process, and that the main regulation occurs at the level of the gene expression.

Overcoming Cellular Immunity to Prolong Adenoviral-Mediated Gene Expression in Sciatic Nerve.

In a previous report, we demonstrated that a first generation (E1- and E3-deleted) recombinant adenovirus can transduce expression of the E. coli lacZ gene into Schwann cells, both in vitro and in vivo, suggesting that this method might be useful for future therapy of peripheral neuropathy, including CMT1. Adenoviral-mediated gene transfer was limited, however, by demyelination and Wallerian degeneration at the site of virus injection, as well as by attenuation of viral gene expression over time. In our current work we have optimized adenoviral-mediated gene expression after intraneural injection into sciatic nerve. Using an improved injection protocol, peak expression of lacZ occurs between 10 and 14 days after injection of 2-week-old animals, decreases thereafter, and there is minimal associated tissue injury. In contrast, very few adenoviral-infected Schwann cells are found in nerves of adult animals 10 days after injection, probably due to immune clearance of viral-infected cells. Consistent with this notion, high levels of lacZ are found in sciatic nerve 30 days after injection of adult SCID mice, which have a genetic defect in both cellular and humoral immunity, of adult ß2 microglobulin-deficient mice (ß2 M -/-), which have a genetic defect in cellular immunity, or of adult mice treated with the immunosuppressing agent FK506. In addition, adenoviral-infected Schwann cells co-cultured with axons in vitro, in the absence of a host immune response, ensheath axons and express lacZ for at least 8 weeks. These data thus demonstrate that expression of first generation recombinant adenovirus in sciatic nerve in adult mice, as in other tissues, is limited mainly by the host cellular immune response to the virus, which can be overcome by attenuation of host cell-mediated immunity. Adenoviral vectors might thus be used to modulate Schwann cell gene expression in patients with peripheral neuropathy after appropriate immunosuppression.