To explore the protective effects of orphan nuclear receptor subfamily 2 group E member 1 (NR2E1) overexpression on nerve regeneration in dentate gyrus (DG) of diabetic rats complicated with depression.

The rat model of diabetes complicated with depression was established by injecting streptozotocin into the tail vein combined with chronic unpredictable mild stimulation. The successful rats were randomly divided into model, control and experimental groups, with 8 rats per group; another 8 normal rats were set as normal group. Control group was injected adeno-associated control virus 4 μL. Experimental group was injected with adeno-associated virus of overexpress NR2E1 4 μL; model and normal groups were injected with 0.9% NaCl. After 14 days, the depression-like behavior of rats was detected by Morris water maze, the fasting blood glucose level was detected by blood glucose meter, and the serum insulin (INS), interleukin -1β (IL-1β) and tumor necrosis factor-α (TNF-α) levels in DG area were detected by enzyme-linked immunosorbent assay. The expression of thymidine analogue bromodeoxyuridine (Brdu) in DG region was detected by immunofluorescence method, and the signal transduction expression levels of microRNA-let-7d (miRNA-let-7d)/NR2E1/silencing information regulatory factor 1 (SIRT1) gene were detected by real-time fluorescence quantitative polymerase chain reaction.

The space exploration time of experimental, control, model and normal groups was (31.25±9.94), (25.50±8.02), (19.75±5.82) and (43.87±12.77) s; the fasting blood glucose were (19.28±3.60), (25.26±5.25), (23.71±4.13) and (4.45±0.40) mmoL·L^-1; the insulin levels were (77.15±3.03), (80.53±4.93), (83.31±5.77) and (48.15±5.18) ng·mL^-1; the IL-1β levels were (28.15±4.68), (36.83±6.97), (40.18±4.40) and (19.15±2.67) pg·mL^-1; the TNF-α levels were (281.15±25.15), (324.06±16.85), (337.08±23.11) and (219.06±27.68) pg·mL^-1; the integrated optical density of Brdu were 1.70±1.19, 0.89±0.12, 0.93±0.14 and 2.14±0.15; the relative expression levels of miRNA-let-7d were 2.02±0.18, 2.67±0.54, 2.88±0.30 and 1.11±0.16; the relative expression levels of NR2E1 mRNA were 2.87±0.39, 0.96±0.09, 1.07±0.12 and 3.29±0.35; the relative expression levels of SIRT1 mRNA were 1.62±0.37, 0.99±0.08, 1.04±0.10 and 2.30±0.34, respectively. Compared with the model group, the above indexes in experimental group have statistical significance (all P<0.05).

The overexpression of NR2E1 receptor in isolated nucleus may protect the dentate gyrus nerve regeneration in diabetic rats with depression by regulating miRNA-let-7d/TLX/SIRT1 signal transduction.

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 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 explore the effect of Wuzi Yanzong pill on testicular dysfunction in subacute aging rats through metabolomics.

Forty-two rats were subcutaneously injected with 200 mg·kg^-1 D-galactose (D-Gal) to induce a subacute aging model. After successful modeling, the rats were randomly divided into four groups: model group, control group, exprimental-L group, exprimental-H group, each with 9 rats. Eight normal rats without D-Gal treatment were used as the normal group. The exprimental-L,-H groups were administered Wuzi Yanzong pill at doses of 0.54 and 2.16 g·kg^-1 by gavage, respectively, while the control group was given 300 mg·kg^-1 metformin by gavage. The normal and model groups were treated with an equal volume of distilled water by gavage. All rats were dosed once daily for 4 weeks. During this period, the each group (except for the normal group) was continued injected 200 mg·kg^-1 D-Gal injections. The levels of testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in serum were measured using enzyme-linked immunosorbent assay (ELISA). The relative expression of proteins involved in testicular secretory function [hydroxysteroid 17-beta dehydrogenase 3 (HSD17B3), cytochrome P450, family 11, subfamily A, polypeptide 1 (CYP11A1), steroidogenic acute regulatory protein (STAR)] was determined by Western blot. Metabolomics analysis was performed to study metabolic changes in the testicular tissue.

The serum T levels in the normal, model, control, experimental-L, and experimental-H groups were (8.67±1.71), (1.95±0.46), (6.50±1.15), (2.45±0.49) and (5.81±0.96) pg·mL^-1, respectively; the FSH levels were (28.53±4.16), (46.73±3.72), (30.18±5.17), (35.35±1.90) and (30.31±4.35) mU·mL^-1, respectively; the LH levels were (22.66±3.97), (37.89±4.16), (24.28±3.75), (37.09±3.98) and (28.35±4.38) mU·mL^-1, respectively; the relative protein expression levels of HSD17B3 were 1.04±0.05, 0.29±0.14, 0.78±0.18, 0.71±0.13 and 0.85±0.18; the relative protein expression levels of CYP11A1 were 1.23±0.15, 0.62±0.08, 1.01±0.05, 0.77±0.15 and 1.24±0.09; the relative protein expression levels of STAR were 0.97±0.12, 0.21±0.05, 0.79±0.19, 0.62±0.23 and 0.96±0.25, respectively. The model group showed significant differences in these indicators when compared with the control and experimental-H groups (all P<0.05). Metabolomics analysis identified six differential metabolites in the experimental-H group compared to the model group. These metabolites were mainly involved in steroid hormone biosynthesis, ovarian steroidogenesis, lipid metabolism, and biotin metabolism.

In vivo and metabolomics studies show that Wuzi Yanzong pill improves antioxidant capacity in the testes, reduces oxidative stress damage, delays aging, and effectively treats testicular dysfunction, thereby improving testicular function.

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

To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of propofol medium/long chain fat milk in Beagle dogs plasma, and to investigate the pharmacokinetics and bioequivalence of the reference preparation of propofol medium/long chain fat milk injection and the subject preparation after intravenous injection in Beagle dogs.

The protein precipitation method was used for pretreatment, and the chromatographic column was Inert Sustain AQ-C₁₈ column (50.0 mm×2.1 mm, 5 μm). Mobile phase: 0.1% ammonia-ammonium acetate aqueous solution, flow rate: 0.4 mL·min^-1, column temperature: 40 ℃; the injection volume was 10 μL; ion source: electrospray ion source, negative ion mode, multi-reaction detection. The specificity, standard curve and lower limit of quantitation, precision and recovery, stability and reproducibility of the method were investigated.

The linear range of propofol was 0.05-10.00 μg·mL^-1, the lower limit of quantitation was 0.05 μg·mL^-1, the accuracy was 94.2%-105.0%, the intra-day and inter-day precision was less than 20%, and the stability was good. The C_max of the reference preparation and the test preparation were (8.75±2.66) and (8.15±2.78)μg·mL^-1 and the AUC_last were (1.71±0.45) and (1.88±0.42) h·μg·mL^-1, respectively.

A simple, sensitive and accurate LC-MS/MS method is established for the determination of propofol.

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 establish a qualification evaluation system for primary investigators (PI) in clinical trials.

Based on the preliminary determination of PI qualification evaluation indicators in accordance with laws and regulations, 19 experts engaged in clinical trials or clinical trial management fields were selected for 2 rounds of Delphi expert consultation, and the weights of each indicator were calculated using principal component analysis.

The positive coefficients for the two rounds of expert consultation were 100.00% and 94.74%, and the expert authority coefficients were 0.940 and 0.933, the coordination coefficients of expert opinions were 0.318 and 0.613. After 2 rounds of expert consultation, the PI qualification evaluation indicators were determined, including 3 primary indicators and 17 secondary indicators. The results of principal component analysis showed that the weight range of each index was 0.037 9 to 0.080 3.

The preliminary construction of the PI qualification evaluation system based on the "PI Qualification Evaluation Form" has strong scientificity and applicability, and can provide references for the evaluation and selection of clinical trial PI.