To investigate the effect of inhibition of mitochondrial pyruvate carrier on Okada-induced cuprotosis in SH-SY5Y cells.

SH-SY5Y cells were induced by okada acid (OA) to establish a model of Alzheimer’s disease (AD). The cells were divided into control group (complete medium), model group (20 nmol·L^-1 OA) and experimental group [20 nmol·L^-1 OA+1 μmol·L^-1 2-cyano-3 -(1-phenyl-1H-indole-3-yl) -2-acrylic acid (UK-5099)]. Copper ion content was detected by cell cupric colorimetric kit. Pyruvate content was detect by pyruvate detection kit, and the expressions of phosphorylated Tau (AT8) and ferredoxin 1 (FDX1) in each group were detected by immunofluorescence staining. The expression levels of mitochondrial pyruvate carrier (MPC1/MPC2) and dihydrolipoyl transacetylase (DLAT) were detected by Western blot.

The contents of copper ions in experimental group, model group and control group were (0.36±0.01), (0.90±0.05) and (0.23±0.04) μmol·g prot^-1; the pyruvate content were (0.55±0.04), (0.33±0.03) and (0.44±0.04) μmol·10⁸ cell^-1; the AT8 expression levels were 91.46±14.53, 125.02±14.43 and 72.57±19.81; the FDX1 expression levels were 98.93±7.40, 147.83±18.63 and 79.18±8.06; the relative expression levels of MPC1 protein were 0.59±0.11, 1.01±0.14 and 0.55±0.13; the relative expression levels of MPC2 protein were 0.54±0.05, 1.31±0.07 and 0.43±0.07; the relative expression levels of DLAT protein were 0.72±0.04, 0.92±0.07 and 0.53±0.14, respectively. The above indexes in the experimental group were statistically significant compared with those in the model group (P<0.05, P<0.01, P<0.001).

Inhibition of mitochondrial pyruvate carrier can alleviate Okada-induced cuprotosis of SH-SY5Y cells.

To explore the effects of 1% lidocaine on pain threshold in rats with myofascial pain syndrome (MPS) based on cyclic adenosine monophosphate (cAMP) / protein kinase A (PKA)/peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) signaling pathways.

The models of MPS rats were prepared by striking manipulation + fatigue method, and they were randomly divided into model group, combined group, experimental-L group and experimental-H group, 10 rats in each group. Another 10 rats were selected as blank group. After successful molding, experimental-L group and experimental-H group were given injection of 1% lidocaine (0.25 mL, 0.5 mL) at pain trigger site of right medial thigh (once a week), combined group was given intraperitoneal injection of SR-18292 (30 mg·kg^-1, 3 times a week) on basis of experimental-H group, model group and blank group were given injection of normal saline (once a week) at pain trigger site of right medial thigh or at the same site. Five groups were treated for 3 weeks. The pain threshold was detected by mechanical stimulation and thermal stimulation, level of adenosine triphosphate (ATP) in muscle tissues at pain trigger site was detected by the kit, and expressions of cAMP, PKA and PGC-1α proteins were detected by Western blot.

After 3 weeks of intervention, paw withdrawal mechanical threshold (PWMT) values in experimental-H group, combined group, model group and blank group were (51.69±7.22), (37.84±6.29), (20.33±5.83) and (67.88±8.46) g, paw withdrawal latency (PWL) values were (10.58±1.03), (7.89±0.87), (6.33±0.92) and (12.89±1.64) s, ATP levels were (866.37±98.66), (728.81±89.60), (617.52±89.84) and (1 098.45±101.36) μmol·g^-1, relative expression levels of cAMP protein were 0.53±0.08, 0.48±0.07, 0.21±0.06 and 0.68±0.10, relative expression levels of PKA protein were 0.60±0.06, 0.58±0.08, 0.28±0.05 and 0.79±0.09, relative expression levels of PGC-1α protein were 0.78±0.05, 0.59±0.06, 0.41±0.08 and 0.93±0.09, and the differences were statistically significant between experimental-H group, combined group and mode group (all P<0.05).

1% lidocaine can improve pain threshold and mitochondrial energy metabolism in muscle tissues of affected limbs in MPS rats, which may be related to activating cAMP/PKA/PGC-1α signaling pathways.

To achieve overexpression of phosphoglycerate kinase 1 (PGK1) in HEK-293T cells and screen the key site of its succinylation through constructing PGK1 overexpression vector (PGK1-WT) and four lysine site succinylation mutation vectors.

GEPIA2 database was used to analyze the expression of PGK1 mRNA in normal breast tissue (291 cases) and breast cancer tissue (135 cases) and the relationship between PGK1 expression level and the overall survival rate of breast cancer patients. Protein modification proteomics was used to detect the levels of protein succinylation in four triple negative breast cancer (TNBC) cell lines (MDA-MB-436, MDA-MB-468, MDA-MB-231, BT-549) and normal breast epithelial cells (MCF-10A); the succinylated positive and negative mutation vectors of K11, K131, K323, K361 were contructed with point mutation kit. The experiment was divided into NC, shPGK1-496, shPGK1-220, PGK1-WT groups and positive and negative mutant groups of K11, K131, K323 and K361. Real-time fluorescence quantitative polymerase chain reaction was used to detect the level of expression of PGK1 mRNA in NC, shPGK1-496 and shPGK1-220 groups. The expression levels of PGK1 proteins in NC, shPGK1-496 and shPGK1-220 groups, and the expression levels of Flag proteins in NC, PGK1-WT and four sites in positive and negative mutant groups were detected by Western blot. The key site of PGK1 succinylation was detected by co-immunoprecipitation.

Compared with normal breast tissues, the level of PGK1 mRNA was highly expressed in breast cancer tissue, and the survival time of breast cancer patients with high PGK1 expression was shorter. Four lysine sites of PGK1 in TNBC cells were hypersuccinylated. The relative expression levels of PGK1 mRNA in NC, shPGK1-496 and shPGK1-220 groups were 1.03±0.23, 0.15±0.11 and 1.42±0.69, respectively; the relative expression levels of PGK1 protein were 1.08±0.05, 0.49±0.05 and 1.28±0.01 respectively; compared with NC group, the above indexes in shPGK1-496 group were statistically different (all P<0.01). The relative expression levels of Flag protein in NC group, PGK1-WT group and positive and negative mutant groups of K11, K131, K323 site were 0, 3.43±0.09, 3.28±0.10, 2.28±0.11, 3.03±0.03, 2.79±0.11, 3.29±0.09 and 2.98±0.04, respectively; compared with the NC group, PGK1-WT group and the positive and negative mutant groups of K11, K131 and K323 had statistical differences (all P<0.01). The relative expression levels of Flag protein in NC group, PGK1-WT group and K361 positive and negative mutant group were 0, 2.58±0.11, 3.04±0.02 and 4.04±0.10, respectively; compared with NC group, PGK1-WT group and K361 positive and negative mutant group had statistical significance (all P<0.01). The levels of lysine succinylation in IgG group, PGK1-WT group and negative mutant groups of K11, K131, K323, K361 were 0, 1.53±0.12, 1.30±0.18, 1.29±0.09, 1.23±0.03 and 0.89±0.16, respectively; compared with PGK1-WT group, the negativ mutant group of K361 had statistical significance (P<0.01).

The four lysine sites of PGK1 in TNBC cells are highly succinylated, and K361 site is critical to their succinylation.

To investigate whether the supplemented Buyang Huanwu tang can improve the kidney injury in diabetic kidney disease (DKD) mice through regulating mitochondrial dynamics.

The db/db mice were randomized into the model group (0.2 mL·d^-1 distilled water), low and high-dose experimental groups (16.0, 32.0 g·kg^-1 supplemented Buyang Huanwu tang), with 8 mice in each group. Another 8 db/m mice were included in the normal group (0.2mL·d^-1 distilled water). The four groups were orally administered once a day for 8 consecutive weeks. Then, the levels of fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), blood urea nitrogen (BUN), serum creatinine (SCr), and urinary microalbumin (mALB) were detected at the end of the experiment. The expression of mitofusin 1 (Mfnl), mitofusin 2 (Mfn2), optic atrophy 1 (Opa1), dynamin-related protein 1 (Drp1), fission protein 1 (Fis1), sirtuin 3 (SIRT3), transforming growth factor-β₁ (TGF-β₁), α-smooth muscle actin (α-SMA), collagen type Ⅲ (Col-Ⅲ) and fibronectin (FN) was measured by Western blotting.

The levels of TG in the experimental-L, -H groups, model group and normal group were (1.98±0.13), (1.69±0.13), (3.38±0.59) and (1.25±0.16) mmol·L^-1; the levels of TC were (3.86±0.14), (3.19±0.15), (5.41±0.61) and (1.46±0.19) mmol·L^-1; the levels of BUN were (10.04±2.21), (8.79±1.75), (16.92±4.57) and (4.78±1.07) mmol·L^-1; the levels of SCr were (19.17±2.25), (17.16±2.03), (27.92±4.13) and (13.37±1.77) μmol·L^-1; the levels of mALB were (26.92±4.61), (20.54±2.31), (46.78±7.32) and (6.14±1.13) mg·L^-1; the relative expression levels of TGF-β1 were 0.73±0.09, 0.59±0.05, 1.01±0.14 and 0.48±0.05; the relative expression levels of α-SMA were 0.79±0.09, 0.56±0.05, 1.12±0.11 and 0.55±0.04; the relative expression levels of FN were 0.61±0.03, 0.55±0.05, 0.83±0.09 and 0.22±0.03; the relative expression levels of Col-Ⅲ were 0.53±0.06, 0.35±0.05, 0.73±0.09 and 0.17±0.03; the relative expression levels of SIRT3 were 0.65±0.07, 0.41±0.14, 0.23±0.12 and 0.91±0.04; the relative expression levels of Drp1 were 0.54±0.08, 0.49±0.09, 0.98±0.11 and 0.37±0.03; the relative expression levels of Fis1 were 0.71±0.05, 0.51±0.03, 1.14±0.11 and 0.35±0.05; the relative expression levels of Mfn1 were 0.61±0.04, 0.95±0.05, 0.43±0.05 and 1.02±0.07; the relative expression levels of Mfn2 was 0.67±0.06, 0.89±0.09, 0.52±0.03 and 1.08±0.05; the relative expression levels of OPA1 were 0.77±0.06, 0.79±0.09, 0.52±0.03 and 1.13±0.11, respectively. Significant differences were found between the model group and experimental-L, -H groups (P<0.05, P<0.01). Simultaneously, the renal pathological and mitochondrial damage was significantly improved.

The supplemented Buyang Huanwu tang can maintain the balance of mitochondrial dynamics, thereby reducing urinary protein, alleviating renal injury and fibrosis. Its potential mechanism may be related to the regulation of SIRT3 protein expression.

To explore the effects of ginsenoside Rh2 on the development of colon cancer through microRNA-128-3p(miR-128-3p)/growth differentiation factor 15(GDF15) mediated wingless-type MMTV integration site family member (Wnt)/β-catenin signaling pathway.

SW620 colorectal cancer cells were randomly divided into the following groups: Control group (standard culture), experimental group (treated with ginsenoside Rh2 at 20 μmol·L^-1), NC inhibitor group (transfected with NC inhibitor), miR-128-3p inhibitor group (transfected with miR-128-3p inhibitor), ginsenoside Rh2 + miR-128-3p inhibitor group (treated with ginsenoside Rh2 at 20 μmol·L^-1 and transfected with miR-128-3p inhibitor), and ginsenoside Rh2+miR-128-3p inhibitor+si-GDF15 group (treated with ginsenoside Rh2 at 20 μmol·L^-1, transfected with miR-128-3p inhibitor and si-GDF15). Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of miR-128-3p and GDF15 mRNA. Cell proliferation was assessed using 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay. Western blotting was employed to detect the protein relative levels of myelocytomatosis oncogene (Myc) and β-catenin.

The relative expression levels of miR-128-3p in the control group and experimental group were 1.00±0.16 and 1.56±0.25, respectively; the relative expression relative levels of GDF15 mRNA were 1.00±0.12 and 0.43±0.05, respectively; the proliferation rates were (88.76±8.56)% and (35.89±4.05)%, respectively; the v-myc avian myelocytomatosis viral oncogene homolog (Myc) protein levels were 1.00±0.21 and 0.45±0.06, respectively; and the β-catenin protein relative levels were 1.00±0.23 and 0.52±0.07, respectively. Compared with the control group, the above indicators in the experimental group showed statistically significant differences (all P<0.05). In the ginsenoside Rh2+miR-128-3p inhibitor group and ginsenoside Rh2+miR-128-3p inhibitor+si-GDF15 group, the cell proliferation rates were (68.56±7.12)% and (50.56±5.18)%, respectively; the Myc protein relative levels were 0.86±0.12 and 0.61±0.08, respectively; and the β-catenin protein relative levels were 0.71±0.09 and 0.58±0.08, respectively. The above indicators in the ginsenoside Rh2+miR-128-3p inhibitor group compared with the experimental group, and those in the ginsenoside Rh2+miR-128-3p inhibitor+si-GDF15 group compared with the ginsenoside Rh2+miR-128-3p inhibitor group, all showed statistically significant differences (all P<0.05).

Ginsenoside Rh2 can inhibit the proliferation and migration of colon cancer SW620 cells, and promote the apoptosis, which may be related to the regulation of ginsenoside Rh2 on the miR-128-3p/GDF15 to inhibit Wnt/β-catenin signaling pathway.

To investigate the effects of paeoniflorin on insulin resistance and myocardial injury in rats with gestational diabetes mellitus and its mechanism.

Female rats were randomly divided into control group (normal diet), model group [male rats fed with high-fat diet and intraperitoneally injected 30 mg·kg^-1 streptozotocin (STZ) after close cages], experimental group (after modeling, 50 mg·kg^-1 paeoniflorin was given by gavage), si-NC group (50 mg·kg^-1 paeoniflorin + si-NC plasmid injected into tail vein after modeling), si-IGFBP2 group (50 mg·kg^-1 paeoniflorin + si-IGFBP2 plasmid injected into tail vein after modeling), each group of 10 rats. After 2 weeks of continuous treatment, fasting blood glucose (FBG) was measured with a glucose meter. Serum insulin-like growth factor binding protein 2 (IGFBP2), fasting insulin (FINS), cardiac troponin Ⅰ (cTnⅠ), creatine kinase isoenzyme MB (CK-MB) related factors were detected by enzyme-linked immunosorbent assay (ELISA), insulin resistance index (HOMA-IRI) was calculated, and the expression of signal pathway-related proteins was detected by Western blot assay.

The levels of FBG in control group, model group, experimental group, si-NC group and si-IGFBP2 group were: (5.62±0.56), (18.10±1.76), (9.81±1.46), (10.02±1.38) and (16.32±1.87) mmol·L^-1, respectively; IGFBP2 levels were (39.47±2.64), (20.65±1.86), (27.18±1.85), (27.95±1.74) and (15.28±1.22) ng·mL^-1, respectively; FINS levels were (8.72±0.75), (21.11±2.50), (16.88±1.86), (16.06±1.39) and (20.01±2.94) mU·L^-1, respectively; cTnⅠ levels were (406.04±29.98), (788.11±84.43), (513.70±57.62), (526.85±38.03) and (756.51±63.88) pg·mL^-1, respectively; CK-MB levels were (7.98±0.07), (15.15±1.26), (11.59±1.23), (11.37±1.79) and (13.49±1.54) U·L^-1, respectively; HOMA-IRI levels were 2.17±0.26, 16.92±2.12, 6.24±0.95, 6.60±1.10 and 12.83±1.68, respectively; the relative expression levels of phosphorylate-phosphatidylinositol 3-kinase (p-PI3K) protein were 0.68±0.11, 0.39±0.04, 0.53±0.05, 0.51±0.06 and 0.43±0.04, respectively; the above indexes in the model group were compared with the control group, the experimental group were compared with the model group, and the si-IGFBP2 group were compared with the si-NC group, the differences were statistically significant (all P<0.05).

Paeoniflorin may improve insulin resistance and myocardium injury in gestational diabetes mellitus rats by up-regulating IGFBP2 expression.

To evaluate the impact of stage assessment nursing mode on the psychological state of healthy participants in Phase I clinical trials of new drugs based on psychological questionnaire.

This was a randomized, single blind, controlled clinical research. Healthy participants were randomly divided into treatment and control group. According to the impact of anxiety and depression on healthy participants, the treatment group was given a stage evaluation nursing model, while the control group received routine nursing measures. Both groups were evaluated using the self rating depression scale (SDS) and self rating anxiety scale (SAS) questionnaires before administration, on the day of administration, after administration (mid-term of the trial), on the day of discharge, and on the day of withdrawal, and their effectiveness was observed.

A total of 159 healthy participants were included in the study, with 83 cases in the treatment group and 76 cases in the control group. SDS scores of treatment group and control group were (54.93±4.62) and (54.87±4.89) points before administration, (52.17±6.55) and (53.95±7.03) points on the administration day, (49.94±7.35) and (51.69±9.16) points at the mid-term of the trial, (52.13±6.70) and (53.85±6.76) points on the discharge day, (51.33±7.99) and (52.60±7.39) points on the day of withdrawal, respectively. Compared with before administration, there was statistically significant difference in treatment group during administration day and the day of withdrawal, as well as in the mid-term of trial in the control group (all P<0.01). SAS scores for treatment group and control group were (36.08±6.45) and (37.04±6.87) points before administration, (36.18±7.79) and (37.43±8.53) points on administration day, (35.04±7.09) and (36.86±9.55) points at the mid-term of the trial, (35.82±7.75) and (37.68±8.78) points on discharge day, (39.29±9.17) and (36.47±7.91) points on the day of withdrawal, respectively. No statistical significance of SAS scores was observed (all P>0.05).

Use of psychological questionnaires and stage nursing assessment forms for evaluation and nursing intervention can alleviate the levels of depression and anxiety in healthy participants.

To evaluate the bioequivalence of test preparation (T) and reference preparation (R) of diphenidol hydrochloride tablets taken orally in a single fasting/postprandibular state in healthy Chinese subjects.

This protocol was designed as random, open, single-center, single-dose, double-cycle and double-cross experiment was selected in this study. A total of 66 healthy male and female subjects were enrolled, including 24 subjects in the test group and 42 subjects in the reference group. All subjects took the test preparation and the reference preparation 25 mg in each cycle. The plasma concentration of diphenidol hydrochloride was determined by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and the pharmacokinetic (PK) data collected was analyzed and processed statistically.

The PK parameters of diphenidol hydrochloride tablets and reference preparations in fasting group were as follows: C_max were (75.90±34.60) and (67.96±19.90) ng·mL^-1, respectively; AUC_0-t were (387.67±198.48) and (341.57±137.05) ng·h·mL^-1, respectively; AUC_0-∞ were (413.00±228.39) and (361.70±161.19) ng·h·mL^-1, respectively. Main PK parameters of diphenidol hydrochloride tablets and reference preparations orally taken by subjects after meals: C_max were (92.60±33.60) and (85.30±31.96) ng·mL^-1, respectively; AUC_0-t were (511.90±206.87) and (498.33±197.78) ng·h·mL^-1, respectively; AUC_0-∞ were (543.21±236.90) and (513.98±203.92) ng·h·mL^-1, respectively. Under fasting and postprandial conditions, subjects took diphenidol hydrochloride tablet and reference preparation orally, one tablet 25 mg each time. The geometric mean of the main pharmacokinetic parameters of difenidol in plasma (C_max, AUC_0-t, AUC_0-∞) and the ratio of their corresponding parameters ranged from 80.00% to 125.00% with 90% confidence interval, and no serious adverse events occurred in both groups.

Diphenidol hydrochloride tablets show bioequivalence and good safety evaluation under fasting and postprandial conditions