Effects of cytoreductive surgery combined with hyperthermic perfusion chemotherapy on prognosis of patients with advanced gallbladder cancer.

BACKGROUND: Gallbladder cancer (GC) is a common malignant tumor and one of the leading causes of cancer-related death worldwide. It is typically highly invasive, difficult to detect in the early stages, and has poor treatment outcomes, resulting in high mortality rates. The available treatment options for GC are relatively limited. One emerging treatment modality is hyperthermic intraperitoneal chemotherapy (HIPEC). HIPEC involves delivering heated chemotherapy directly into the abdominal cavity. It combines the strategies of surgical tumor resection and localized chemotherapy administration under hyperthermic conditions, aiming to enhance the concentration and effectiveness of drugs within the local tumor site while minimizing systemic toxicity. AIM: To determine the effects of cytoreductive surgery (CRS) combined with HIPEC on the short-term prognosis of patients with advanced GC. METHODS: Data from 80 patients treated at the Punan Branch of Renji Hospital, Shanghai Jiao Tong University School of Medicine between January 2018 and January 2020 were retrospectively analyzed. The control group comprised 44 patients treated with CRS, and the research group comprised 36 patients treated with CRS combined with HIPEC. Then, the survival time and prognostic factors of the two groups were compared, as well as liver and kidney function indices before and six days after surgery. Adverse reactions and complications were recorded in both groups. RESULTS: The baseline data of the research and control groups were similar (P > 0.05). Six days after surgery, the alanine aminotransferase, aspartate aminotransferase, total bilirubin, and direct bilirubin levels significantly decreased compared to the preoperative levels in both groups (P < 0.05). However, the values did not differ between the two groups six days postoperatively (P > 0.05). Similarly, the postoperative creatinine and blood urea nitrogen levels were significantly lower than the preoperative levels in both groups (P < 0.05), but they did not differ between the groups six days postoperatively (P > 0.05). Furthermore, the research group had fewer postoperative adverse reactions than the control group (P = 0.027). Finally, a multivariate Cox analysis identified the tumor stage, distant metastasis, and the treatment plan as independent factors affecting prognosis (P < 0.05). The three-year survival rate in the study group was higher than that in the control group (P = 0.002). CONCLUSION: CRS combined with HIPEC lowers the incidence of adverse reactions and improves survival in patients with advanced GC.

Association of lymphopenia and RDW elevation with risk of mortality in acute aortic dissection.

OBJECTIVE: The study aimed to investigate whether lymphopenia and red blood cell distribution width (RDW) elevation are associated with an increased risk of mortality in acute aortic dissection (AAD). METHODS: This multicenter retrospective cohort study enrolled patients diagnosed with AAD by aortic computed tomographic angiography (CTA) from 2010 to 2021 in five teaching hospitals in central-western China. Cox proportional hazards regression and Kaplan-Meier curves were used in univariable and multivariable models. Clinical outcomes were defined as all-cause in-hospital mortality, while associations were evaluated between lymphopenia, accompanied by an elevated RDW, and risk of mortality. RESULTS: Of 1903 participants, the median age was 53 (interquartile range [IQR], 46-62) years, and females accounted for 21.9%. Adjusted increased risk of mortality was linearly related to the decreasing lymphocyte percentage (P-non-linearity = 0.942) and increasing RDW (P-non-linearity = 0.612), and per standard deviation (SD) of increment lymphocyte percentage and RDW was associated with the 26% (0.74, 0.64-0.84) decrement and 5% (1.05, 0.95-1.15) increment in hazard ratios (HRs) and 95% confidence intervals (CIs) of mortality, respectively. Importantly, lymphopenia and elevation of RDW exhibited a significant interaction with increasing the risk of AAD mortality (P-value for interaction = 0.037). CONCLUSIONS: Lymphopenia accompanied by the elevation of RDW, which may reflect the immune dysregulation of AAD patients, is associated with an increased risk of mortality. Assessment of immunological biomarkers derived from routine tests may provide novel perspectives for identifying the risk of mortality.

Modulation of Cerium Carbonate Crystal Growth by Polyvinylpyrrolidone using Density Functional Theory.

Cerium carbonate crystal morphology is predicted using density functional theory (DFT) simulations in this paper. In the nucleation phase, the ketone group in polyvinylpyrrolidone (PVP) will preferentially bind to Ce3+ to form complexes and provide heterogeneous nucleation sites for the system, prompting the nucleation of cerium carbonate crystals. In the growth stage, due to the adsorption of PVP, the probability of (120) crystal plane appearing in the equilibrium state is the greatest, resulting in the formation of hexagonal flake cerium carbonate crystals with (120) crystal plane as the oblique edge. Experimentally, hexagonal sheet cerium carbonate crystals were successfully prepared using PVP as a template agent. Therefore, DFT can be used to predict the morphology of cerium carbonate crystals, which not only elucidates the growth mechanism of cerium carbonate crystals, but also greatly reduces the experimental cost.

High Level of Palmitic Acid Induced Over-Expressed Methyltransferase Inhibits Anti-Inflammation Factor KLF4 Expression in Obese Status.

Our study is based on the establishment of a cohort of human obese omental adipose tissue and the culture of adipocytes in vitro. To observe the effect of high level of free fatty acid (FFA) on the expression of DNA methyltransferases (DNMTs) and the anti-inflammatory factor Kruppel-like factor 4 (KLF4) in adipocytes and evaluate the role of methyltransferases in FFA inhibiting KLF4 expression. A total of 20 normal patients and 20 obese patients were selected for further test. qRT-PCR and western blot were used to detect the mRNA and protein expression levels of DNMT1/DNMT3a/DNMT3b and KLF4 in human adipose tissue and 3T3-L1 adipocytes which stimulated with saturated fatty acid, palmitic acid (PA). Bisulfite sequencing PCR (BSP) detected methylation status of KLF4 gene in human adipose tissue. It was found that the mRNA and protein expression levels of DNMT1 and DNMT3a in the omental tissue of obese individuals were higher than those in normal group, but the expression of KLF4 was decreased. The positive methylation rate of KLF4 promoter region in obese individuals were significantly higher than those in normal individuals, especially at CpG_33 and CpG_34 sites. Meanwhile compared with non-methylated group at CpG_33 and CpG_34 sites of KLF4 promoter region, the DNMT3a mRNA expression in methylated group were significantly increased. A total of 200 µM PA significantly promoted DNMT1, DNMT3a, and DNMT3b and inhibited KLF4 protein expression levels in 3T3-L1 adipocytes. Our findings suggest that under obesity status, the lower expression level of KLF4 of visceral adipose tissue may correlate with palmitic acid promoted DNMTs expression in adipocytes.

Young and old adipocytes have differential influence on the development of osteoblasts.

OBJECTIVE: The aim of the current study was to investigate the effect of adipocytes on the differentiation of osteoblasts at different stages of adipocyte development. METHODS: BMSCs were isolated from 4-week-old male wistar rat femurs and tibias, and flow cytometry was performed. Adipocytes were derived from BMSCs, cell morphology was continually observed from day 21 to day 50. Adipocyte medium was collected once every 2days (d) and ELISA kits were used for detection of triglycerides (TG), tumor necrosis factor-α (TNF-α), and interleukin-6(IL-6) expression level. 21d and 40d old adipocyte and osteoblast cells were co-cultured, and alizarin red staining was performed after 21d. After co-culture, the adherent cells were collected, and the expression of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) was detected by real time PCR. RESULT: Results of cell characterisation showed that the cells had positive expression of CD29 (97.92%) and CD44 (89.32%). With the increase of the induction time of mature adipocytes, the number of adipocyte on 21thd was significantly higher than 40thd, while the volume of adipocyte was significantly lower than 40thd (P<0.05). The levels of TG(2.6±0.83mmol/l VS 3.8±0.66mmol/l), TNF-α(30.5±2.53pg/ml VS 57.6±5.1pg/ml), and IL-6(32.5±1.42pg/ml VS 55.1±5.97pg/ml) secreted by adipocytes increased with induction time: 40thd was significantly higher than 21thd (P<0.01). When 21thd adipocytes and osteoblasts were co-cultured, the number of calcium nodules significantly increased over that of the positive control group, When 40thd adipocytes and osteoblasts were co-cultured, the number of calcium nodules significantly decreased over that of the positive control group (P<0.05). The OPG(68.9±5.39 VS 1.00±0.36) expression was significantly increased, and the expression of RANKL (2.0±0.84 VS 34.4±2.01) was significantly decreased from the 21thd adipocytes co-cultured group compared with the 40thd adipocytes co-cultured group (P<0.001). CONCLUSION: The differential size of adipocytes in the bone marrow can affect bone metabolism by regulating the expression of OPG/RANKL.

The Effect and Mechanism of KLF7 in the TLR4/NF-κB/IL-6 Inflammatory Signal Pathway of Adipocytes.

OBJECTIVE: To investigate the role and possible molecular mechanism of Krüppel-like factor 7 (KLF7) in the TLR4/NF-κB/IL-6 inflammatory signaling pathway activated by free fatty acids (FFA). METHODS: The mRNA and protein expression levels of KLF7 and the factors of TLR4/NF-κB/IL-6 inflammatory signal pathways were detected by qRT-PCR and Western blotting after cell culture with different concentrations of palmitic acid (PA). The expression of KLF7 or TLR4 in adipocytes was upregulated or downregulated; after that, the mRNA and protein expression levels of these key factors were detected. KLF7 expression was downregulated while PA stimulated adipocytes, and then the mRNA and protein expressions of KLF7/p65 and downstream inflammatory cytokine IL-6 were detected. The luciferase reporter assay was used to determine whether KLF7 had a transcriptional activation effect on IL-6. RESULTS: (1) High concentration of PA can promote the expression of TLR4, KLF7, and IL-6 in adipocytes. (2) TLR4 positively regulates KLF7 expression in adipocytes. (3) KLF7 positively regulates IL-6 expression in adipocytes. (4) PA promotes IL-6 expression via KLF7 in adipocytes. (5) KLF7 has a transcriptional activation on IL-6. CONCLUSION: PA promotes the expression of the inflammatory cytokine IL-6 by activating the TLR4/KLF7/NF-κB inflammatory signaling pathway. In addition, KLF7 may directly bind to the IL-6 promoter region and thus activate IL-6.

The Effect and Mechanism of TLR9/KLF4 in FFA-Induced Adipocyte Inflammation.

OBJECTIVE: Current research has reported that obesity is a chronic inflammatory state, which is closely related with excessive accumulation of free fatty acid, while the specific mechanism that high level of FFA causes inflammation is not very clear. Thus, our research intended to observe the high FFA effects on TLR9/KLF4 expression and the downstream inflammatory factors, to explore the mechanism of inflammatory response suppressed by TLR9/KLF4. METHODS: qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of TLR9, KLF4, and key inflammation-related factors. ELISA was used to detect the release level of inflammatory cytokines. The triglyceride (TG) and glucose (GLU) testing cassettes were used to detect the TG and GLU levels in culture medium. RESULTS: In the omental tissue of obese individuals (OB), we found that TLR9, KLF4, mRNA, and the protein expression levels were lower than those of the normal weight control (NC) group. Similarly, in the omental tissue of high-fat diet (HFD) rats, we found that the mRNA expression levels of TLR9 and KLF4 were lower than those of the normal diet control group. In mature adipocytes, we found that KLF4 played an important anti-inflammatory role; moreover, PA can promote the development of inflammation by inhibiting KLF4 expression; TLR9 has a positive regulation function on KLF4 expression, but unrelated to PA. CONCLUSIONS: TLR9/KLF4 is involved in regulating FFA-induced adipocyte inflammation.

Contrast-enhanced ultrasonography assessment of gastric cancer response to neoadjuvant chemotherapy.

AIM: To quantitatively assess the ability of double contrast-enhanced ultrasound (DCUS) to detect tumor early response to pre-operative chemotherapy. METHODS: Forty-three patients with gastric cancer treated with neoadjuvant chemotherapy followed by curative resection between September 2011 and February 2012 were analyzed. Pre-operative chemotherapy regimens of fluorouracil + oxaliplatin or S-1 + oxaliplatin were administered in 2-4 cycles over 6-12 wk periods. All patients underwent contrast-enhanced computed tomography (CT) scan and DCUS before and after two courses of pre-operative chemotherapy. The therapeutic response was assessed by CT using the response evaluation criteria in solid tumors (RECIST 1.1) criteria. Tumor area was assessed by DCUS as enhanced appearance of gastric carcinoma due to tumor vascularity during the contrast phase as compared to the normal gastric wall. Histopathologic analysis was carried out according to the Mandard tumor regression grade criteria and used as the reference standard. Receiver operating characteristic (ROC) analysis was used to evaluate the efficacy of DCUS parameters in differentiating histopathological responders from non-responders. RESULTS: The study population consisted of 32 men and 11 women, with mean age of 59.7 ± 11.4 years. Neither age, sex, histologic type, tumor site, T stage, nor N stage was associated with pathological response. The responders had significantly smaller mean tumor size than the non-responders (15.7 ± 7.4 cm vs 33.3 ± 14.1 cm, P < 0.01). According to Mandard's criteria, 27 patients were classified as responders, with 11 (40.7%) showing decreased tumor size by DCUS. In contrast, only three (18.8%) of the 16 non-responders showed decreased tumor size by DCUS (P < 0.01). The area under the ROC curve was 0.64, with a 95%CI of 0.46-0.81. The effects of several cut-off points on diagnostic parameters were calculated in the ROC curve analysis. By maximizing Youden's index (sensitivity + specificity - 1), the best cut-off point for distinguishing responders from non-responders was determined, which had optimal sensitivity of 62.9% and specificity of 56.3%. Using this cut-off point, the positive and negative predictive values of DCUS for distinguishing responders from non-responders were 70.8% and 47.4%, respectively. The overall accuracy of DCUS for therapeutic response assessment was 60.5%, slightly higher than the 53.5% for CT response assessment with RECIST criteria (P = 0.663). Although the advantage was not statistically significant, likely due to the small number of cases assessed. DCUS was able to identify decreased perfusion in responders who showed no morphological change by CT imaging, which can be occluded by such treatment effects as fibrosis and edema. CONCLUSION: DCUS may represent an innovative tool for more accurately predicting histopathological response to neoadjuvant chemotherapy before surgical resection in patients with locally-advanced gastric cancer.

[Effects of no-tillage and stubble-remaining on soil enzyme activities in broadcasting rice seedlings paddy field].

A field experiment was conducted to study the effects of four cultivation modes (conventional tillage, no-tillage, conventional tillage + stubble-remaining, and no-tillage + stubble-remaining) on the activities of urease, acid phosphatase, protease, and cellulose in different soil layers in a broadcasting rice seedlings paddy field. Under the four cultivation modes, the activities of test enzymes were higher in upper than in deeper soil layers, and had a greater difference between the soil layers under no-tillage + stubble-remaining. In upper soil layers, the activities of test enzymes were higher in the treatments of no-tillage than in the treatments of conventional tillage, being the highest under no-tillage + stubble-remaining and the lowest under conventional tillage. In deeper soil layers, the test enzyme activities were the highest under conventional tillage + stubble-remaining, followed by no-tillage + stubble-remaining, no-tillage, and conventional tillage. During the growth period of rice, soil urease and cellulose activities were lower at tillering stage, increased to the maximum at booting stage, and decreased then, soil acid phosphatase activity was higher at tillering stage but lower at elongating stage, whereas soil protease activity peaked at tillering and heading stages.

[Study on the interaction of Sm(III) complexes of rutin with serum albumin].

The binding reaction of rutin-Sm with serum albumin (SA) was investigated by the fluorescence method in physiological condition. The authors studied mainly the quenching mechanism of the fluorescence of SA by rutin-Sm, and calculation of the binding constants K(LB) of human serum albumin (HSA) and bovine serum albumin (BSA) with rutin-Sm by Lineweaver-Burk equation at different temperatures respectively, then obtained the thermodynamic parameters of HSA and BSA with rutin-Sm according to the calculated binding constants K(LB) at different temperature, meanwhile the type of binding forces of HSA and BSA with rutin-Sm was determined. The results showed that the emission spectra of BSA (HSA) in the presence and absence of rutin-Sm are different. The emission spectra of BSA (HSA) in the presence of rutin-Sm can be quenched. The quenching mechanism of rutin-Sm to SA was static quenching with non-radiation energy transfer for new complex of SA and rutin-Sm. The binding constants K(LB) (L x moL(-1)) were 6.540 x 10(5) and 3.265 x 10(5) for BSA, and 6.830 x 10(5) and 4.665 x 10(5) for HSA at 295 K and 310 K respectively. And the type of bonding forces was estimated by the calculation of thermodynamic parameters of the reaction of rutin-Sm with SA at different temperatures, and the result showed that the binding forces were mainly H-bond and Van der Waals between BSA and rutin-Sm due to the deltaH < 0 and deltaS < 0, and the main electrostatic interaction of rutin-Sm and HSA because of deltaH < 0 and deltaS > 0. The effect of rutin-Sm on the conformation of serum albumin was also studied by using synchronous fluorescence spectroscopy. Results indicated that rutin-Sm could be deposited and transported by serum protein in vivo.

[Effects of returning straw to soil and different tillage methods on paddy field soil fertility and microbial population].

A field experiment was conducted on a paddy field to study the effects of returning straw to soil and different tillage methods (no-tillage + returning straw, no-tillage, tillage + returning straw, and tillage) on the fertility level and microbial quantities of different soil layers. The results showed that in upper soil layer, the organic matter content in treatment 'no-tillage + returning straw' was 5.33, 2.79, and 5.37 g x kg(-1) higher than that in treatments 'no-tillage', 'tillage + returning straw', and 'tillage', respectively, and the contents of total and available N, P and K in treatment 'no-tillage + returning straw' were also the highest, followed by in treatments 'no-tillage' and 'tillage + returning straw', and in treatment 'tillage'. In deeper soil layer, all the fertility indices were higher in treatment 'tillage + returning straw'. Treatments of 'returning straw to soil' had the highest quantities of soil microbes. The quantities of bacteria, fungi, and actinomycetes in upper soil layer were the highest in treatment 'no-tillage + returning straw', and thus, the cellulose decomposition intensity in this treatment at maturity period was 26.44%, 79.01%, and 98. 15% higher than that in treatments 'tillage + returning straw', 'no-tillage', and 'tillage', respectively. In deeper soil layer, the quantities of bacteria, fungi, and actinomycetes were the highest in treatment 'tillage + returning straw'. Treatment 'no-tillage + returning straw' had the features of high fertility and abundant microbes in surface soil layer. The quantities of soil bacteria and actinomycetes and the decomposition intensity of soil cellulose were significantly positively correlated with soil fertility level.

[Thermodynamics studies on the binding of rutin and serum albumin].

To study the binding of rutin and serum albumin (SA) in physiological condition and the quenching mechanism of the fluorescence of SA by rutin, the fluorescence method was used. The results shows that the emission spectra of BSA (HSA) in the presence and absence of rutin are different. The emission spectra of BSA (HSA) in the presence of rutin can be quenched. The quenching mechanism of rutin to SA was static quenching with non-radiation energy transfer with single molecule. The binding constants K(A), the number of binding sites n and the thermodynamic parameters of the reaction of rutin with SA were determined at different temperatures. At 295 and 310 K, for BSA, K(A)(L * moL(-1)) = 4.215 x 10(4) and 6.996 x 10(3) and n = 0.75 and 0.64, respectively; for HSA, K(A)(L * moL (-1)) = 2.660 x 10(4) and 4.110 x 10(3) and n = 0.70 and 0.60, respectively. The binding constants K(A) decreased with the increase in temperature, which means that rutin and SA have a quite strong ability to form a new complex-system. The main binding force was discussed by thermodynamic equation, and the result is that deltaH < 0 and deltaS < 0 for the reaction of rutin with SA. So the binding forces was mainly H-bond and Van der Waals. The effect of the drug on the conformation of serum albumin was also studied by using synchronous fluorescence spectroscopy. Rutin could be deposited and transported by serum protein in vivo. Rutin had nearly no effect on the serum protein conformation.

[Study on the interactions of Eu(III) complex of O-(thioxanthone-[2]-yl)-oxyacetic acid with DNA].

The interaction of the Eu(III) complex of O-(thioxanthone-[2]-yl)-oxyacetic acid and calf thymus DNA was studied using fluorimetric, UV-Vis and CD(circular dichroism) methods. It was found that the ultraviolet absorption intensity of Eu(III) complex of O-(thioxanthone-[2]-yl)-oxyacetic acid decreased as the concentration of DNA increased under the physiological condition of pH 7.10, and the compound remarkably influenced the negative peak in CD spectra of DNA when Eu(III) complex of O-(thioxanthone-[2]-yl)-oxyacetic acid interacted with DNA. The emission intensity of the Eu(III) complex evidently increased in the presence of DNA. The emission intensity of the DNA-EB system decreased as the concentration of the complex increasesd. And the presence of EB also led to the decrease in the emission intensity of the DNA-complex system. These changes observed here are often characteristic of intercalation. The above results strongly suggest that the complex can bind to DNA mainly by intercalation.

[Influence of rutin on conformation of serum albumin].

We studied the effect of the rutin on the conformation of bovine serum albumin (BSA) and human serum albumin (HSA) by synchronous fluorescence spectrum (Alambda = lambda(em) - lambda(ex) = 15 nm and deltalambda = lambda(em) -lambda(ex) = 60 nm) and circular dichroism spectra. The results showed that rutin had hardly effect on the serum protein conformation, but influenced the secondary structure of serum albumin (SA) molecule with the addition of rutin into BSA and HSA solution. The alpha-helix structure of BSA and HSA was decreased and the beta-sheet was increased with the increase in rutin concentration. At the same time, we studied the binding of rutin and bovine serum albumin (BSA) and human serum albumin (HSA) by electrochemistry under physiological condition. Cyclic voltammograms of rutin demonstrated a pair of well-reversible peaks in the solution of Tris-NaCl buffer at pH 7.38 (sweep rate: 50 mV x s(-1); a glassy carbon working electrode, a platinum auxiliary electrode, and a saturated calomel reference electrode). Both peak potentials of rutin were E(pc) = 0.103 2 V and E(pa) = 0.150 6 V, ipc : ipa = 1 : 1.2, the dispersion of peak potential was 47.4 mV (deltaE(prutin) = deltaE(pc) - E(pa) = 47.4 mV). In addition, with the addition of BSA and HSA into the rutin solution, both the reduction and oxidation currents decreased only at the peak potentials of rutin (BSA: E(pc) = 0.114 1 V, E(pa) = 0.168 5 V, ipc : ipa = 1 : 1.2; HSA: E(pc) = 0.114 2 V, E(pa) = 0.168 8 V, ipc : ipa= 1 : 1.1), the dispersions of peak potential were changed: for BSA deltaE(prutin) = [E(pc) - E(pa)] = 54.4 mV, and for HSA: deltaE(prutin) = [E(pc) - E(pa)] = 54.6 mV. The results showed that there was an interaction of rutin with BSA and HSA, forming a kind of nonelectroactive supramolecular complex, and indicated that rutin could be deposited and transported by serum protein in vivo.