Aim To investigate effects of indole-3-carbinol (I3C) on hepatic stellate cells (HSCs) activated by acetaldehyde in precision-cut liver slices (PCLS). Methods PCLS were incubated with 700 ?mol?L-1 acetaldehyde and 200 ~ 800 ?mol?L-1 I3C for 6 h. The expression of ?-smooth muscle actin(?-SMA) in liver slices was analyzed by immunohistochemistry. The leakages of glutathione S-transferase (GST), lactate dehydrogenase (LDH) and content of transforming growth factor-?1 (TGF-?1) in media were assayed. Contents of malondialdehyde (MDA) and hydroxyproline (Hyp) in tissue were also determined. Expressions of matrix metalloproteinases-1 (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1) in media were analyzed by the western blot. Results The increase of activated HSCs due to acetaldehyde was inhibited by I3C (200~800 ?mol?L-1). Meanwhile, I3C treatment (200~800 ?mol?L-1) showed significant and concentration-dependent antagonistic actions on the increment of GST, LDH leakages into the media and MDA, Hyp contents in tissues induced by acetaldehyde. The increase of TGF-?1 was also remarkable inversed by I3C (200~800 ?mol?L-1). As compared with acetaldehyde group, the ratio of MMP-1/TIMP-1 was increased significantly by I3C treatment (800 ?mol?L-1)(P

AIM: To activate hepatic stellate cells (HSC) in vitro in precision-cut liver slices (PCLS) stimulated by acetaldehyde for studying and screening anti-fibrotic drugs. METHODS: PCLS were prepared by the vibratome, and incubated with 700 ?m?L~-1 acetaldehyde for 0, 2, 4 and 6 h. The medium and homogenate were retained to determine glutathione S-transferase (GST) activity, lactate dehydrogenase (LDH) leakage and hydroxyproline (Hyp) content. PCLS were prepared for paraffin sections. The expression of ?-smooth muscle actin (?-SMA) was evaluated by immunohistochemistry, and the result was analyzed with image analysis software. RESULTS: The leakages of GST and LDH were increased significantly compared with those in 0 h group (P

To study the effect on regulation of cell cycle of osteosarcoma cell line MG63 tranceduced with exogenous p16ink4a and hRb1 genes, pIRES-p16ink4a-hRb1, pIRES-p16ink4a and pIRES-hRb1 plasmids were constructed by gene recombination technology. The recombinant plasmid was transferred into osteosarcoma cell line MG63 by metafectene, and the resistant clones were selected by G418 selective medium. mRNA and protein expression of osteosarcoma cell line were assayed by RT-PCR and Western Blot respectively. Cell cycle and apoptosis were analyzed by subG1 flow cytometric. Cell proliferation was tested by MTT. In the genome of these transfected target cells, the expression of p16ink4a and hRb1 mRNA and protein were detected respectively in vitro. It was demonstrated with subG1 flow cytometric analysis and MTT method that p16ink4a and hRb1 genes cooperation more significantly inhibited cell growth and induced a more marked G1 arrest and apoptosis than p16ink4a/hRb1 alone (P＜0.01). Coexpression of exogenous p16ink4a with hRb1 broke the regulatory feedback loop of p16ink4a-cyclinD1/CDK-hRb1 and played a more significant role in inhibiting cell growth as well as inducing cell apoptosis than p16ink4a or hRb1 did alone in vitro.

Objective:To evaluate the long-term safety, tolerability and efficacy of Lacosamide add-on therapy in Chinese children with partial-onset seizures.Methods:SP848 was a global multicenter single-arm study involving 60 Chinese children with partial-onset seizures with the age of 4-17 years who were managed by Lacosamide add-on therapy at seven hospitals across China from April 2018 to May 2019.After treatment with at least two kinds of anti-seizure medications simultaneously or sequentially, partial seizures were still poorly controlled and Lacosamide oral solution (syrup) or tablets were added.The minimum initial oral dose was 2 mg/(kg·d), and the maximum allowable dose was 12 mg/(kg·d)or 600 mg/d during the study period.The dose was adjusted according to the tolerance and seizure control level of partial-onset seizures children.Seizure frequency and the median percentage change in partial-onset seizures per 28 days from baseline to the final visit were recorded, including 50% responder rate and 75% responder rate.Results:A total of 60 Chinese children with the mean age of 9.18 (4.00-15.40) years were included in this interim analysis, involving 39 males and 21 females.The mean course of epilepsy was 5.04 (0.50-15.20) years.A total of 43 patients (71.7%) still have been treated.One patient (1.7%) has completed the 6-12 months of follow-up, and 14 patients (23.3%) have completed the follow-up for less than 6 months.The median change in the frequency of partial seizures every 28 days from baseline to the last visit was -2.91, with its median percentage as -25.46%, and the proportions of ≥50%, while ≥75% responder rate were 40.0% and 28.3%, respectively.A total of 52 patients (86.7%) had 265 treatment emergent adverse events (TEAE), 11 patients (18.3%) had 19 serious TEAE, 37 patients (61.7%) had 127 drug-related TEAE, and 11 patients (18.3%) had 16 TEAE leading to the discontinuation of the trial.The most common TEAE were upper respiratory tract infections (20 cases, 33.3%), followed by drowsiness (16 cases, 26.7%), dizziness (15 cases, 25.0%) and vomiting (13 cases, 21.7%). There were no abnormal changes in the electrocardiographic findings during the treatment.Conclusions:For Chinese patients with partial seizures who are older than the age of 4 years and poorly controlled by other drugs, Lacosamide is effective and well tolerated as an add-on therapy drug.The safety characteristics are consistent with those reported in children and adults.No new safety concerns are identified.

Objective:To observe central responses and functional connectivity (FC) during urinary bladder storage in neurogenic overactive bladder (NOAB).Methods:Twenty patients with NOAB were recruited. Resting state functional magnetic resonance imaging (rs-fMRI)were performed in all subjects under the following two conditions: empty bladder state and full bladder (strong desire to void)state. Software MATLAB, SPM8 and DPABI were adopted to analyze the difference of brain imaging between the two conditions. Voxel-based analysis of the REHO was performed to analyze rs-fMRI data including the main excitatory regions and inhibitory areas, peak value(X-axis, Y-axis, Z-axis), clusters size (active volume unit: number of voxel), T value(the excitatory and inhibitory extent of brain active regions). Voxel-based analysis of the REHO maps and FC between empty and full bladder were performed.Results:Increased activity during strong desire to void with NOAB patients was observed in the left orbital part of superior frontal, right middle frontal gyrus, and right superior frontal. Decreased activity was observed in right precentral. FC analysis found that these activated or deactivated brain regions were widely connected with other brain areas, include: frontal lobe, parietal lobe, temporal lobe, cingulate gyrus, lenticular nucleus, insular lobe, angular gyrus, parahippocampal gyrus and anterior and posterior central gyrus.Conclusions:Our results suggested that the right frontal robe may play a role in the control of bladder with NOAB during strong desire to void, and inhibitory areas located in right precentral. In NOAB patients, excitatory and inhibitory connections were increased in frontal lobe and central gyrus, decreased in insular lobe and parhippocampal gyrus.

ObjectiveTo investigate the effect and possible mechanism of Shenfu Injection （参附注射液） on rat cardiomyocyte injury induced by isoproterenol from the perspective of regulating the high mobility group protein B1 （HMGB1）/ Toll-like receptor 4 （TLR4）/nuclear factor κB （NF-κB） pathway through the deacetylation modification of silent information regulator 1 （SIRT1）. MethodsThe optimal concentration and intervention duration of isoproterenol hydrochloride and the optimal intervention concentration of Shenfu Injection were screened out by CCK-8 method. Logarithmically growing H9c2 rat cardiomyocytes were taken at 5×10⁴ cells/well and divided into normal group， model group， Shenfu Injection group， and SIRT1 inhibitor group， with 3 replicates in each group.Except for the normal group， the cells in the other groups were induced by isoproterenol hydrochloride to establish a chronic heart failure cell model. After modeling， the Shenfu Injection group was given Shenfu Injection at the optimal intervention concentration， and the SIRT1 inhibitor group was given 1 μmol/L of SIRT1 inhibitor EX-527， for optimal intervention duration.CCK-8 assay was used to detect the cell activity and calculate the inhibitory rate. ELISA assay was used to detect the nicotinamide adenine dinucleotide oxidation state/ nicotinamide adenine dinucleotide reduction state （NAD+/NADH） in cardiomyocytes. Immunofluorescence was used to detect the immunofluorescence localization of HMGB1 and SIRT1 in cardiomyocytes. Western blotting was used to detect the protein expression of acetylated HMGB1 in cardiomyocytes， HMGB1 in the nucleus and cytoplasm， and SIRT1， TLR4， myeloid differentiation factor 88 （MYD88） and NF-κB p65 in cardiomyocytes. RT-qPCR was used to detect the mRNA expression of SIRT1， HMGB1， TLR4， MYD88 and NF-κB p65 in cardiomyocytes. ResultsThe optimal intervention concentration of isoproterenol hydrochloride was 300 μmol/L， and the intervention duration was 48 hours； 8% was the optimal intervention concentration of Shenfu Injection. Compared to those in the normal group， the cell activity， NAD+/NADH value， nuclear HMGB1 protein expression， cardiomyocyte SIRT1 protein and mRNA expression in the model group decreased， while the cell inhibition rate， cardiomyocyte acetylated HMGB1 and cytoplasmic HMGB1 protein expression， cardiomyocyte TLR4， MYD88， NF-κB p65 protein and mRNA expression all increased （P＜0.05）； fluorescence localization showed that the content of HMGB1 in cardiomyocytes in the model group increased and was localized in both the nucleus and cytoplasm.Compared to the model group and the SIRT1 inhibitor group， the Shenfu Injection group showed significant improvements in all the above indicators （P＜0.05）； fluorescence localization showed that the SIRT1 content increased in the Shenfu injection group， while the HMGB1 content decreased， and was mainly located in the nucleus. ConclusionShenfu Injection can improve myocardial cell damage by increasing SIRT1 expression to reduce the acetylation level of HMGB1， regulating the HMGB1/TLR4/NF-κB pathway and inhibiting the nuclear translocation of HMGB1.

Objective:To explore the efficacy and safety of intravesical electrical stimulation (IVES) combined with a training for bladder motor and sensory dysfunction in the treatment of neurogenic underactive bladder(UAB).Methods:A prospective, single-blind, randomized controlled trial was used to study neurogenic UAB patients admitted to the China Rehabilitation Research Center from October 2019 to May 2021. Inclusive criteria included age≥18 years old, the patients who have been diagnosed as neurogenic UAB and the course of disease being more than 3 months; patients who have been undergone intermittent catheterization to empty the bladder or patients indicated for intermittent catheterization (post-void residual urine accounts for more than 40% of the functional bladder volume), voluntary signing of written informed consent, able to communicate well with researchers and comply with the requirements of the whole trial, and the patient not undergoing any treatment other than oral medication before IVES. Exclusion criteria included patients with low bladder compliance by urodynamic examination(<20 ml/cmH 2O), patients with mechanical outflow obstruction, patients with complete spinal cord injury, the patients with symptomatic urinary tract infection which was not cured, patients with hydronephrosis or bladder-ureteral reflux, patients with renal insufficiency(serum creatinine greater than 1.5 times of the upper limit of normality), patients with malignant tumors of the bladder or prostate, overactive bladder, Alzheimer's disease, brain atrophy, acute cerebrovascular disease, or cognitive impairment, patients who were pregnant or planning to be pregnant, bladder mucosa injury, patients with pacemakers or defibrillators, those who participated in other clinical trials 3 months before the study, and other circumstances that the researcher consider it is not suitable to be involved in this study. The patients were randomly divided into experimental group and control group according to the ratio of 1∶1. The experimental group used conventional transurethral insertion of bipolar catheter electrodes for IVES combined with bladder motor and sensory dysfunction training, and the control group underwent IVES with open circuit combined with bladder motor and sensory dysfunction training. The stimulation parameters of the two groups were two-way square wave, 1-30 mA intensity, 10-20 Hz frequency, 200 μs pulse width, once a day, lasting 30 minutes for each treatment, and for continuous 20 working days. The post-void residual urine, voiding efficiency, 24-hour intermittent catheterization times, first sensation of bladder filling volume and American Urological Association Symptom Index Quality of Life(AUA-SI-QOL) scores were recorded before and at the end of treatment. The adverse events during the treatment were recorded. Results:Fifty-two patients were selected and 50 patients completed the trial, including 26 patients in the experimental group and 24 patients in the control group. Before treatment, there were no significant differences in gender[16(male)/10(female)vs.13(male)/11(female), P=0.598], age [(40.7±13.5)years vs.(38.5±12.3)years, P=0.543], course of disease[0.71(0.42, 1.63)years vs.0.79(0.42, 1.50)years, P=0.695], post-void residual urine[300(193, 400)ml vs.325(178, 380)ml, P=0.724], voiding efficiency[17%(0, 47.8)% vs.21%(0, 38.0)%, P=0.960], 24-hour intermittent catheterization times[4(2, 4)vs.3(2, 4), P=0.692], first sensation volume during bladder filling[(325.8±74.3)ml vs.(307.5±75.0)ml, P=0.391] or AUA-SI-QOL scores[5(4, 5)vs.4(4, 5), P=0.313] between the experimental group and the control group. At the end of treatment, the post-void residual urine, first sensation volume during bladder filling and AUA-SI-QOL scores of the experimental group were significantly lower than those of the control group [250(40, 350)ml vs.300(200, 390)ml, P=0.034; (276.5±68.8)ml vs.(315.4±67.3)ml, P=0.049; 4(2, 4)vs.4(3, 5), P=0.024], and the voiding efficiency was significantly higher than that of the control group[33%(14.5, 84.5)% vs.18%(0, 35.8)%, P=0.041], but there was no significant difference in the number of 24-hour intermittent catheterization between the two groups [3(1, 4)vs.3(2, 4), P=0.174]. In the control group, there were no significant changes in post-void residual urine, voiding efficiency, 24-hour intermittent catheterization times, first sensation volume during bladder filling and AUA-SI-QOL scores before and after treatment [325(178, 380)ml vs.300(200, 390)ml, P=0.832; 21%(0, 38.0)% vs.18%(0, 35.8)%, P=0.943; 3(2, 4)vs.3(2, 4), P=0.239; (307.5±75.0)ml vs.(315.4±67.3)ml, P=0.257; 4(4, 5)vs.4(3, 5), P=0.157]. In the experimental group, there were significant improvements in post-void residual urine, voiding efficiency, 24-hour intermittent catheterization times, first sensation volume during bladder filling and AUA-SI-QOL scores before and after treatment [300(193, 400)ml vs.250(40, 350)ml, P<0.001; 17%(0, 47.8)% vs.33%(14.5, 84.5)%, P<0.001; 4(2, 4)vs.3(1, 4), P=0.011; (325.8±74.3)ml vs.(276.5±68.8)ml, P<0.001; 5(4, 5)vs.4(2, 4), P<0.001]. During the treatment period, 1 case of abdominal discomfort occurred in the experimental group and 1 case of urethral discomfort in the control group. After adjusting the stimulation intensity and catheter position, the discomfort disappeared without other serious adverse events. Conclusions:IVES combined with bladder motor sensory dysfunction training can not only effectively improve the bladder emptying efficiency and bladder sensation in patients with neurogenic UAB, but also be safe and easy to operate.

Chronic heart failure is a serious heart disease with dyspnea and limited activity tolerance as the main clinical manifestations. Activation of the inflammatory system can significantly stimulate cardiac fibrosis and remodeling and promote the progression of heart failure， playing a key role in the development of the disease. Studies have confirmed that inflammation is involved in the development of different types of heart failure. "Toxic pathogen theory" is an important basic theory of traditional Chinese medicine （TCM） to explain the occurrence of diseases. We concluded the similarities between TCM toxic pathogens and inflammation in concept， disease location， etiology， syndrome differentiation， and clinical characteristics. Chronic heart failure is manifested by the toxic pathogens of turbid phlegm， stagnated blood， and accumulated fluid. Heart vessel obstruction is the main pathological factor， and the inflammatory factors produced by necrotic cardiomyocytes are the microscopic manifestations of the obstruction. Therefore， based on the "toxic pathogen theory"， this study aimed to clarify the role of inflammation in the development of chronic heart failure from both macroscopic and microscopic perspectives. Moreover， this paper proposed that the stagnated blood has not been transformed into toxin in the early stage of the disease and thus the products of clearing heat and detoxification should not be used. At the development stage of the disease when the transformation tends to begin， treatment should be based on syndrome differentiation， and detoxifying Chinese medicine should be used in order to achieve the goal of "removing toxin without harming the healthy Qi". At the late stage of heart failure， toxins have been accumulated and detoxifying medicines and therapies should be applied to eliminate the toxic pathogens. This study is expected to lay a foundation for the modern research on the role of inflammation in the development of chronic heart failure with TCM theory and guide the diagnosis and treatment of this disease.

For those patients with refractory lower urinary tract dysfunction who are not well treated by traditional therapy such as behavior therapy and drug therapy, neuromodulation technologies have gradually become alternative treatments. Several neuromodulation technologies are also used in animal experimental and clinical scientific research by more and more scholars, in order to find more effective methods and mechanisms of treatment of lower urinary tract dysfunction. This article introduces the principle and advantages of common neuromodulation technologies, which focuses on the application in lower urinary tract dysfunction treatment, and analyzes the direction and the broad prospect of neuromodulation.
Humans ; Transcutaneous Electric Nerve Stimulation ; Urologic Diseases ; physiopathology ; therapy

ObjectiveTo study the modeling characteristics of primary dysmenorrhea models in animals and to provide references for the standardization of the primary dysmenorrhea animal models. MethodThe research articles on animal models of primary dysmenorrhea were retrieved to establish a database. The types of experimental animals， modeling methods， modeling cycle， drug dosage， drug injection methods， high-frequency detection indicators， positive drug types， etc.， were summarized and analyzed. ResultA total of 171 research articles that met the criteria were included. The animals for primary dysmenorrhea model induction were mainly SD rats， Wistar rats， and Kunming mice. Most of them were prepared by combining estradiol and oxytocin with the modeling cycle of 9 d≤t≤12 d. In terms of drug dosage for rats， estradiol benzoate was 0.5 mg·d^-1 on the 1st and 10th days and 0.2 mg·d^-1 on the 2^nd to 9^th days， while oxytocin at 2 U·d^-1 was the most common. In terms of drug dosage for mice， diethylstilbestrol at 2 mg·kg^-1·d^-1 and oxytocin at 20 U·kg^-1·d^-1 were the most common. In terms of injection methods， oxytocin was mainly administered by intraperitoneal injection and estradiol （estradiol benzoate and diethylstilbestrol） by subcutaneous injection. The detection indicators were mainly behavioral indicators of the writhing assay or the related biochemical indicators in the uterus or serum by enzyme-linked immunosorbent assay. The positive western medicines were dominated by ibuprofen and Chinese medicines by Tongjingbao. ConclusionAlthough primary dysmenorrhea animal models have become a hot topic， the existing reviews are not comprehensive， and the modeling standards and traditional Chinese medicine （TCM） syndrome evaluation are inadequate. By summarizing and analyzing the big data of the animal models， this study proposed some specific views to provide guidance and references for establishing the standard and ideal animal models of primary dysmenorrhea， so as to carry out research on this disease.