Effects of different doses of remimazolam on hemodynamics during general anesthesia in patients with septic shock.

OBJECTIVE: The stability of hemodynamics plays a vital role in the process of anesthesia induction for patients with septic shock. As a new-type benzodiazepine, remimazolam has numerous advantages, including rapid induction, rapid recovery, stable hemodynamics, and mild respiratory depression. Nevertheless, reports about the effects of remimazolam on hemodynamics in patients with septic shock are still limited. The study aimed to evaluate the effects that different doses of remimazolam have on hemodynamics in inducing general anesthesia in patients with septic shock. PATIENTS AND METHODS: Admitted to the intensive care unit of our hospital from January 2023 to June 2023, 75 patients with septic shock caused by acute appendicitis-induced sepsis were selected as observation subjects. They were randomly assigned to receive low-dose [0.2 mg/(kg·h)], medium-dose [0.3 mg/(kg·h)], and high-dose [0.4 mg/(kg·h)] remimazolam by using a random number table, with 25 patients in each group. Their intraoperative conditions were recorded, including operation duration, intraoperative hemorrhage volume, intraoperative transfusion volume, and decannulation time. Hemodynamic parameters, including mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), and stoke volume index (SVI) were collected at seven-time points (T0: before induction; T1: before intubation; T2: after intubation; T3: the start of operation; T4: 15 min after operation; T5: 30 min after operation; T6: the end of operation). We also compared hepatic and renal function indexes, including blood urea nitrogen (BUN), serum creatinine (sCr), procalcitonin (PCT), white blood cells (WBC), tumor necrosis factor-α2 (TNF-α2), and Interleukin-6 (IL-6), of the three groups of patients before operation and 1, 3, 5, 7 days after operation. In addition, the incidence of adverse reactions in the three groups was recorded and compared. RESULTS: During remimazolam induction, the number of patients with intraoperative need for rescue remimazolam in the medium-dose and high-dose groups was significantly lower than in the low-dose group (p < 0.05). In terms of hemodynamic indexes, MAP in the high-dose group at T2 was lower than that at T0 (p < 0.05), and MAP at T2 was significantly lower in the high-dose group than that in the medium-dose group (p < 0.05). Furthermore, MAP at T4 in the medium-dose and high-dose groups declined compared with the low-dose group (p < 0.05). There were no significant differences in HR, CI, and SVI at different time points among the three groups (p > 0.05), but levels of HR and SVI decreased and CI increased after anesthesia compared with those before operation. Additionally, in comparison with the levels before operation, levels of sCR, BUN, PCT, WBC, TNF-α, and IL-6 were higher on postoperative days 1, 3 (p < 0.05) and lower on postoperative day 7 (p < 0.05). After the operation, both levels of BUN and sCR in the medium-dose and high-dose groups were lower than those in the low-dose group (p < 0.05). CONCLUSIONS: Remimazolam is safe and effective for inducing general anesthesia in patients with septic shock. Low, medium, and high doses of remimazolam can maintain a stable hemodynamic state, and the recovery of hepatic and renal function is certain to depend on the dose.

UCHL1 Impairs Periodontal Ligament Stem Cell Osteogenesis in Periodontitis.

Periodontitis comprises a series of inflammatory responses resulting in alveolar bone loss. The suppression of osteogenesis of periodontal ligament stem cells (PDLSCs) by inflammation is responsible for impaired alveolar bone regeneration, which remains an ongoing challenge for periodontitis therapy. Ubiquitin C-terminal hydrolase L1 (UCHL1) belongs to the family of deubiquitinating enzymes, which was found to play roles in inflammation previously. In this study, the upregulation of UCHL1 was identified in inflamed PDLSCs isolated from periodontitis patients and in healthy PDLSCs treated with tumor necrosis factor-α or interleukin-1ß, and the higher expression level of UCHL1 was accompanied with the impaired osteogenesis of PDLSCs. Then UCHL1 was inhibited in PDLSCs using the lentivirus or inhibitor, and the osteogenesis of PDLSCs suppressed by inflammation was rescued by UCHL1 inhibition. Mechanistically, the negative effect of UCHL1 on the osteogenesis of PDLSCs was attributable to its negative regulation of mitophagy-dependent bone morphogenetic protein 2/Smad signaling pathway in periodontitis-associated inflammation. Furthermore, a ligature-induced murine periodontitis model was established, and the specific inhibitor of UCHL1 was administrated to periodontitis mice. The histological results showed increased active osteoblasts on alveolar bone surface and enhanced alveolar bone regeneration when UCHL1 was inhibited in periodontitis mice. Besides, the therapeutic effects of UCHL1 inhibition on ameliorating periodontitis were verified, as indicated by less bone loss and reduced inflammation. Altogether, our study proved UCHL1 to be a key negative regulator of the osteogenesis of PDLSCs in periodontitis and suggested that UCHL1 inhibition holds promise for alveolar bone regeneration in periodontitis treatment.

[Effect of adenovirus-mediated shRNA down-regulates SHP2 expression on the apoptosis of human hepatic stellate cells LX-2].

Objective: To investigate the effect of adenovirus-mediated short hairpin RNA (shRNA) downregulating SH2 domain-containing protein tyrosine phosphatase 2 (SHP2) on the apoptosis of human hepatic stellate cells LX-2 cultured in vitro. Methods: The recombinant adenovirus Ad-shRNA/SHP2 carrying shRNA targeted SHP2 and expressing green fluorescent protein (GFP), and the empty control virus Ad-GFP expressing GFP were transfected into LX-2 cells cultured in vitro. Real-time fluorescence quantitative PCR was used to detect SHP2 mRNA expression in LX-2 cells. Western blot was used to detect the protein expressions of SHP2, Bax, and Bcl-2 in LX-2 cells. TUNEL and annexin-V/propidium iodide dual-labeled flow cytometry were used to detect apoptosis in LX-2 cells. Experimental group: (1) Control group: LX-2 cells were transfected with DMEM instead of adenovirus; (2) Ad-GFP group: transfected with empty virus Ad-GFP; (3) Ad-shRNA/SHP2 group: transfected with recombinant adenovirus Ad-shRNA/SHP2. The means between multiple groups were compared using a one-way ANOVA and the LSD test was used for inter group comparisons. Results: shRNA-targeted SHP2 significantly down-regulated the expression of SHP2 protein and mRNA in LX-2 cells (P < 0.05). The TUNEL and annexin-V/propidium iodide dual-labeled flow cytometry results showed that the apoptosis rate of LX-2 cells in the Ad-shRNA/SHP2 group (12.755%±1.606%, 19.340%±2.505%) (P < 0.05) was significantly higher compared to the control group (3.077%±0.731%, 9.438%±0.804%) and the Ad-GFP group (3.250%±0.851%, 8.893%±1.982%), with no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). Western blot analysis of Bax and Bcl-2 protein expression in LX-2 cells of each group revealed that the Bax protein expression was significantly higher in the Ad shRNA/SHP2 group (2.493 ± 0.203) (P < 0.05) compared to the control group and Ad-GFP group (1.989 ± 0.147, 1.999 ± 0.162), with no statistically significant difference between the control group and the Ad-GFP group (P > 0.05), while the Bcl-2 protein was significantly decreased in the Ad-shRNA/SHP2 group (1.042±0.148) compared with the control group and the Ad-GFP group (1.707±0.146, 1.521±0.142), with no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). Conclusions: SHP2 expression down-regulation induces apoptosis of human hepatic stellate cells LX-2 in vitro by reducing Bcl-2/Bax.

Comparing the Performance of Different Instruments for Diagnosing Frailty and Predicting Adverse Outcomes among Elderly Patients with Gastric Cancer.

OBJECTIVES: To examine the diagnostic performance of the Tilburg Frailty Indicator (TFI), 11-factor modified frailty index (mFI-11), and 5-factor modified frailty index (mFI-5) for frailty defined by Frailty Phenotype (FP), as well as to compare the predictive ability of TFI, mFI-11, and mFI-5 for adverse outcomes in hospital among elderly patients undergoing gastric cancer surgery. DESIGN: A prospective cohort study. SETTING: Hospitalization setting, Nanjing, China. PARTICIPANTS: We recruited 259 elderly patients undergoing gastric cancer surgery from a tertiary hospital. MEASUREMENTS: Frailty was assessed by the FP, TFI, mFI-11, and mFI-5 before surgery, respectively. The receiver operating characteristic (ROC) curves were plotted to compared the diagnostic performance of TFI, mFI-11, and mFI-5 using FP as the reference. ROC curves were used to examine the performance of TFI, mFI-11, and mFI-5 in predicting adverse outcomes. The area under the curve (AUC)>0.70 was regarded as an indicator of good performance. RESULTS: The prevalence of frailty ranged from 8.5% (mFI-11) to 45.9% (TFI). The AUCs of TFI (AUC: 0.764, p<0.001) was significantly greater than that of mFI-11 (AUC: 0.600, p=0.033) and mFI-5 (AUC: 0.600, p=0.0311) in the detection of frailty defined by FP, with quite different sensitivity and specificity at their original cutoffs. TFI and mFI-11 both had statistically significant but similarly inadequate predictive accuracy for adverse outcomes in hospital, including total complications (AUCs: 0.618; 0.621), PLOS (AUCs: 0.593; 0.639), increased hospital costs (AUCs: 0.594; 0.624), and hypoproteinemia (AUCs: 0.573; 0.600). For the mFI-5, only the predictive ability for hypoproteinemia was statistically significant, with poor accuracy (AUC: 0.592, p<0.0055). CONCLUSION: The TFI performed slightly better than mFI-11 and mFI-5 in our study. Moreover, future studies are needed to further determine an optimal frailty instrument with great diagnostic and predictive accuracy.

[Changes of electroencephalography power spectrum during joint position perception test after anterior cruciate ligament rupture].

OBJECTIVE: To measure the electroencephalography (EEG) of the patients with anterior cruciate ligament (ACL) rupture when performing joint position perception movement task, to compare the differences between the ACL rupture side and the unaffected side, to identify the EEG change in the power spectrum caused by the ACL rupture, and to provide evidence for the diagnosis, treatment and rehabi-litation for ACL injury as well as knee instability. METHODS: Sixteen male patients, selected from the Department of Sports Medicine, Peking University Third Hospital from November 2014 to April 2015, with only ACL rupture on one side used isokinetic muscle strength testing equipment were enrolled in the study to perform unilateral active knee joint positional movement and passive knee joint positional movement tasks. EEG was recorded to compare between the affected and unaffected limb of ACL rupture patients when doing single leg movement tasks, including passive knee joint position test and active knee joint position sensation test. The target position of the active knee joint position movement task and the passive knee joint position movement task was 30 degrees of knee flexion. RESULTS: During the passive knee joint position test, there was no significant difference in EEG power spectrum of Delta[F (1, 15)=0.003, P=0.957, ηP2 =0.001], Theta[F (1, 15)=0.002, P=0.962, ηP2 < 0.001], Alpha[F (1, 15)=0.002, P=0.966, ηP2 =0.001], Beta[F (1, 15)=0.008, P=0.929, ηP2 =0.001] at Fz, Cz, and Pz between the affected and unaffected limbs in the ACL patients. During the active knee joint position movement task, the EEG power spectrum of Delta, Theta, Alpha, Beta at Fz and Cz location, on the affected side was significant higher than on the unaffected side. CONCLUSION: This study compared the differences between the ACL rupture side and the unaffected side during active knee position movement task and passive knee position movement task, and identifyied the EEG changes in the power spectrum caused by the ACL rupture, It was found that the central changes caused by unilateral ACL rupture still existed during contralateral (unaffected) side movement. The EEG power spectrum of the affected side during active exercise was significantly higher than that of the unaffected side This study provides new electrophysiological evidence for the study of ACL injury.

Circ_NCKAP1 promotes skin basal cell carcinoma progression by sponging the miR-148b-5p/HSP90 axis.

OBJECTIVE: Skin basal cell carcinoma (BCC) is the most common malignant skin tumor. Recent studies demonstrated that circular RNAs (circRNAs) are implicated in tumorigenesis and may represent potential therapeutic targets. The aim of the present study was to explore the expression profiles of circRNAs and their role in skin BCC. MATERIALS AND METHODS: Three pairs of skin BCC tissues and adjacent tissues were used to perform a circRNA microarray for screening of circRNA expression profiles. Circ_NCKAP1 was selected as a target circRNA by RT-qPCR verification and bioinformatics analysis. The effect of circ_NCKAP1 knockdown on cell proliferation and apoptosis was assessed using CCK8 and Annexin V-FITC/PI assays, and its regulation over the miR-148b-5p/HSP90 axis was assessed by dual-luciferase reporter assay. RESULTS: Circ_NCKAP1 was found to be significantly upregulated in skin BCC tissues (p<0.05). In vitro loss-of-function assays demonstrated that circ_NCKAP1 knockdown markedly inhibited cell proliferation and promoted cell apoptosis (p<0.05). Moreover, Dual-Luciferase reporter assay identified that circ_NCKAP1 could bind to miR-148b-5p directly, and HSP90 was targeted by miR-148b-5p. CONCLUSIONS: Circ_NCKAP1 can promote skin BCC progression by sponging the miR-148b-5p/HSP90 axis, and circ_NCKAP1 may be a potential target for skin BCC therapy.

Maintenance of cellular annexin A1 level is essential for PI3K/AKT/mTOR-mediated proliferation of pancreatic beta cells.

Annexin A1 (AnxA1, also known as lipocortin-1), is a calcium-dependent phospholipid binding protein with diverse functions. Previous studies have indicated that AnxA1 is associated with age-related ß-cell dysfunction and aging, which lead to decreased ß-cell proliferation capacity. However, it has been uncertain whether AnxA1 affects the proliferation of pancreatic beta (ß) cells. In the present study, we reduced AnxA1 expression in the MIN6 islet ß-cell line using small interfering RNA (AnxA1-siRNA), then measured cell cycle distribution and cellular proliferation. We also measured the expression levels of cell cycle-related proteins such as cyclin D1, cyclin E, and cyclin-dependent kinase 2 (CDK2) by Western blot analysis. We investigated the phosphatidylinositol 3-kinase (PI3K)/ serine/threonine protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway to explore the potential mechanism underlying the observed effects. Knockdown of AnxA1 expression using siRNA reduced the rates of MIN6 cell proliferation. The proportions of cells in S and G2/M phases also decreased upon inhibition of AnxA1. Moreover, AnxA1 protein expression in MIN6 cells was positively related to the protein levels of cyclin D1, cyclin E, and CDK2. Activation of the PI3K/Akt/mTOR signaling pathway by AnxA1 may be involved in the signaling cascade to regulate cell proliferation. This study identified a positive correlation between AnxA1 protein and pancreatic ß-cell proliferation. AnxA1 protein expression might affect the proliferation of MIN6 cells via regulation of cyclin D1, cyclin E, and CDK2 proteins, as well as the PI3K/Akt/mTOR signaling pathway.

The detection of SAS1B in serum provides clues for early diagnosis of thyroid cancer.

OBJECTIVE: The incidence of thyroid cancer is rising globally. Most patients progress slowly, but some patients develop lymph node and distant metastasis earlier, and their prognosis is poor. Therefore, early diagnosis and warning of malignancy are very meaningful for such patients. SAS1B gene is a newly discovered protein expressed on the surface of mature egg cells and has metalloendopeptidase activity. We aimed at exploring whether SAS1B is involved in the occurrence of thyroid cancer, and at providing evidence for early diagnosis and targeted therapy of thyroid cancer. PATIENTS AND METHODS: In this study, a rabbit anti-human SAS1B polyclonal antibody was prepared by gene recombination technology. The indirect ELISA method was used to detect the SAS1B protein expression in the serum of 69 patients with thyroid cancer and 55 normal controls, and the relevant pathological factors were analyzed. Immunohistochemistry and PCR technology were used to investigate the expression levels of SAS1B protein and mRNA in 30 thyroid cancer tissues and 23 control thyroid tissues. RESULTS: The titer of SAS1B recombinant antibody was 1:51200. The expression of SAS1B in the serum of patients with thyroid cancer was higher than that in the normal control group (p<0.01). The antibody had a good sensitivity in serum detection of cancer patients (p=0.008<0.01), the linear regression analysis result was that the expression of SAS1B gene was related to tumor envelope invasion and lymph node metastasis (p=0.003<0.01, p=0.003<0.01), and it was irrelevant to the patient's gender, age, tumor mass size, number of cancer foci, pathological stage, etc. (p>0.05). The results of immunohistochemistry showed that SAS1B protein was mainly located in the cytoplasm and membrane of thyroid cancer cells. The expression intensity in thyroid cancer tissues was higher than that in control tissues (p<0.05), but it was not expressed in normal thyroid tissues. Antibodies showed a good sensitivity that was used to detect thyroid cancer tissues (p=0.000<0.01). The results of ordinary PCR detection using thyroid cancer tissue and control thyroid tissue showed that the amplification products of the three domains (N-terminal, C-terminal and catalytic domain) of the SAS1B gene showed high expression in thyroid cancer tissue. q-PCR results showed that the expression of SAS1B gene in thyroid cancer and control thyroid tissue was higher than that in control group (p<0.05), and the genes of Aurora A and BARD1 related to centrosome replication and DNA replication forks protection during the proliferation were highly expressed in thyroid cancer tissue. The study results suggested that SAS1B was involved in the carcinogenesis of thyroid cancer. The Hum_mPLoc.2.0 software, PSORT Ⅱ software and UniProt software were used to predict that SAS1B protein had secretory protein properties. CONCLUSIONS: The above data indicate that the SAS1B gene is closely related to the process of thyroid cancer and can serve as a good tumor marker that can be used for early diagnosis and early warning of thyroid malignancy.

MiR-665 inhibits inflammatory response in microglia following spinal cord injury by targeting TREM2.

OBJECTIVE: The purpose of this study was to uncover the role of microRNA-665 (miR-665) in protecting inflammatory response in microglia following spinal cord injury (SCI) and the underlying mechanism. PATIENTS AND METHODS: The serum levels of miR-665 and TREM2 (triggering receptor expressed on myeloid 2) in SCI patients (n=24) and healthy subjects (n=24) were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Then, the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). After lipopolysaccharide (LPS) induction in BV2 cells, the relative levels of miR-665 and TREM2 were detected by qRT-PCR, and relative levels of IL-6 and TNF-α in the culture medium were examined by ELISA. Next, TREM2, the target gene of miR-665, was determined by Dual-Luciferase reporter assay, and the relationship between the expression levels of TREM2 and miR-665 in SCI patients and BV2 cells was analyzed. Finally, the regulatory effects of miR-665 and TREM2 on IL-6 and TNF-α levels in the culture medium of LPS-induced BV2 cells were assessed. RESULTS: It was found that miR-665 was downregulated in serum of SCI patients and LPS-induced BV2 cells, while TREM2 was upregulated. Silenced miR-665 or overexpressed TREM2 was involved in protecting inflammatory response following SCI. Besides, rescue experiments showed that miR-665 participated in the regulation of inflammatory response following SCI by targeting TREM2. CONCLUSIONS: MiR-665 inhibits inflammatory response following SCI by targeting TREM2.

[Influence of Halogenated Hydroxyl-Alkanes Inhalation Anesthetic on the Determination of Ethanol Content in Blood].

ABSTRACT: Objective To study the influence of halogenated hydroxyl-alkanes inhalation anesthetic on the determination of ethanol content in blood. Methods Halogenated hydroxyl-alkanes were analyzed by headspace gas chromatography with double column confirmatory detection method. The influence of halogenated hydroxyl-alkanes on determination of ethanol content in blood sample by headspace gas chromatography was explored under the different detection conditions of KB-BAC1/ KB-BAC2 and J&W DB-ALC1/DB-ALC2 gas chromatographic column. Results The retention time of sevoflurane and enflurane was similar to that of ethanol and tert butanol respectively when using the J&W DB-ALC1/DB-ALC2 gas chromatographic column, and interfered with the detection of ethanol content in blood; only J&W DB-ALC1 gas chromatographic column can separate the sevoflurane and ethanol components, so as to eliminate their influence on the detection of ethanol content in blood. When using KB-BAC1/KB-BAC2 gas chromatographic column, the retention time of sevoflurane, isoflurane and ethanol is similar, especially that of sevoflurane and ethanol, and sevoflurane obviously interferes with the determination of ethanol content in blood. Conclusion Halogenated hydroxy-alkanes interfere with determination of ethanol content in blood by headspace gas chromatography. The interference can be discriminated effectively by choosing the suitable chromatographic column and double column confirmatory detection.

[Progress of research on the environmental and genetic influencing factors of colorectal cancer].

Colorectal cancer is a global public health issue which possesses serious challenge. The incidence of colorectal cancer in developed countries and regions stands in the forefront worldwide, and has been rising sharply in some of the developing countries. It is unanimously recognized that the occurrence of colorectal cancer is closely related to environmental factors as diet and lifestyle, genetic factors, and gene-environment interactions of the people. Since there have been many studies on the influencing factors of colorectal cancer, the current review aims at providing evidence on colorectal cancer prevention by evaluating the relationships between the influencing factors and colorectal cancer, based on the published literatures. Environmental risk factors revealed by previous epidemiological studies facilitate the population-based prevention programs against colorectal cancer. The developments of sequencing and omics technologies provide more chances to illustrate the genetic susceptibility of colorectal cancer. With both, we are able to construct more accurate risk prediction models and subsequently developing personalized intervention plans to achieve the ultimate goal of reducing the burden of colorectal cancer.

LncRNA MIR497HG inhibits proliferation and migration of retinal endothelial cells under high-level glucose treatment via miRNA-128-3p/SIRT1 axis.

OBJECTIVE: The aim of this study was to elucidate the potential influence of MIR497HG on regulating proliferative capacity of human retinal endothelial cells (HRECs). MATERIALS AND METHODS: Relative expression levels of MIR497HG, microRNA-128-3p (miRNA-128-3p) and SIRT1 in HRECs treated with different doses of glucose and mannitol were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Dual-Luciferase reporter gene assay was conducted to assess the interaction among MIR497HG, miRNA-128-3p, and SIRT1. In addition, the potential effects of MIR497HG/miRNA-128-3p/SIRT1 axis on proliferative and migratory capacities in HRECs were evaluated by Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'- deoxyuridine (EdU) and transwell assay, respectively. RESULTS: High-level glucose (HG) treatment significantly downregulated MIR497HG and SIRT1 expression, whereas upregulated miRNA-128-3p expression in HRECs (p<0.05). MiRNA-128-3p was the target gene binding MIR497HG, and SIRT1 was the downstream gene of miRNA-128-3p. Overexpression of MIR497HG significantly attenuated proliferative and migratory abilities of HG-induced HRECs (p<0.05). Furthermore, decreased trends were partially reversed by overexpression of miRNA-128-3p or knockdown of SIRT1. CONCLUSIONS: MIR497HG is downregulated after HG treatment. In addition, it suppresses the proliferation and migration of HRECs by targeting miRNA-128-3p/SIRT1 axis, thus influencing the progression of diabetic retinopathy.

Trichostatin A alleviates the process of breast carcinoma by downregulating LPAR5.

OBJECTIVE: To elucidate the role of histone deacetylase inhibitor Trichostatin A (TSA) in affecting metastasis of breast carcinoma, and its molecular mechanism. PATIENTS AND METHODS: LPAR5 levels in breast carcinoma tissues and paracancerous tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR), and its expression pattern was further verified in breast carcinoma cell lines. The relationship between LPAR5 and prognosis of breast carcinoma patients was analyzed. After TSA induction (100-400 nmol/L) for 6-48 h, the proliferative and migratory abilities of SKBR3 and MDA-MB-231 cells in overexpressing LPAR5 were examined by cell counting kit-8 (CCK-8), transwell and wound healing assay. By constructing a xenograft model in nude mice, the influences of TSA and LPAR5 on in vivo growth of breast carcinoma were examined. RESULTS: LPAR5 was upregulated in breast carcinoma samples. High level of LPAR5 predicted higher rates of lymphatic metastasis and distant metastasis, as well as lower overall survival and progression-free survival in breast carcinoma patients. LPAR5 level was dose-dependently downregulated in TSA-induced SKBR3 and MDA-MB-231 cells. In addition, TSA induction dose-dependently declined proliferative ability, and time-dependently attenuated migratory ability in breast carcinoma cells. In vivo overexpression of LPAR5 in nude mice reversed the inhibitory effect of TSA on breast carcinoma growth. CONCLUSIONS: TSA induction can suppress proliferative and migratory abilities in breast carcinoma by downregulating LPAR5.

[Effect of knockdown of O-Glcnac transferase on hepatocyte fat synthesis].

Objective: To investigate the effect of knockdown of O-GlcNAc transferase (OGT) on hepatocyte fat synthesis. Methods: Liver cell line L02 were used to established the model of hepatic steatosis. The levels of OGT and O-GlcNAc protein were detected by Western blot. The OGT knockdown cell line of L02 cells was established, and its lipid formation ability was detected after induction of oleic acid (OA). Real-time quantitative PCR (qRT-PCR) and Western blot were used to detect mRNA and protein expression of enzymes related to fat synthesis. An independent sample t test was used. Results: Western blot showed that the expression of OGT and O-GlcNAc was increased in L02 cells after adipogenesis (P < 0.05). After shOGT lentivirus infects L02 cells, OGT mRNA levels were down-regulated (P < 0.01). Oil red O staining showed that the lipid in L02 shOGT cells decreased, qRT-PCR showed that the mRNA expressions of fat synthase (ACC1), (FASN) and (SCD1) were decreased, the difference was statistically significant (P < 0.05), protein Expression is consistent with mRNA expression. Conclusion: Knockdown of OGT can inhibit hepatocyte fat synthesis by reducing O-GlcNAc levels.

Regulatory mechanism of TGF-ß1/SGK1 pathway in tubulointerstitial fibrosis of diabetic nephropathy.

OBJECTIVE: The aim of this study was to clarify the potential function of transforming growth factor-ß1/serum/glucocorticoid-regulated kinase 1 (TGF-ß1/SGK1) pathway in diabetic nephropathy-induced tubulointerstitial fibrosis. MATERIALS AND METHODS: Type 2 diabetes mellitus (T2DM) model was successfully established in rats by high-sucrose-high-fat diet combined with streptozotocin (STZ) induction. Subsequently, blood glucose level, renal function and pathological changes in kidneys of T2DM and control rats were evaluated. Western blot and quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) were conducted to determine the protein and mRNA expression levels of TGF-ß1, SGK1, fibronectin (FN) and α-smooth muscle actin (α-SMA) in rat kidney tissues, respectively. RESULTS: Blood glucose (BG), glycosylated hemoglobin (GHb), serum creatinine (Scr) and blood urea nitrogen (BUN) in T2DM rats were significantly higher than those of control rats (p<0.05). The morphology of glomeruli and renal tubules in rats of control group were normal. In contrast, T2DM rats showed significant lesions in glomeruli, renal tubules, and renal interstitium. Furthermore, the relative expression levels of TGF-ß1, SGK1, FN, and α-SMA in kidney tissues of T2DM rats were remarkably higher than those of controls (p<0.05). CONCLUSIONS: The TGF-ß1/SGK1 pathway is closely related to tubulointerstitial fibrosis in T2DM rats.