OBJECTIVE To explore the molecular mechanism of Compound Dihuang Granule in treating Parkinson's disease(PD)with yin-deficiency and dynamic wind syndrome by regulating the changes of ubiquitin-proteasome system-related proteins.METHODS SPF male SD rats were randomly selected as normal control group and sham operation group,13 rats in each group.The remaining rats were treated with 6-hydroxydopamine lateral brain injection to damage the substantia nigra to establish PD yin-deficien-cy and dynamic wind syndrome rat model.The established rats were randomly divided into model group,madopar group(150 mg·kg-1),low,medium and high dose groups of Compound Dihuang Granule(1.75,3.5,7 g·kg-1),13 rats in each group.Each group was given drugs by intragastric gavage for 28 d.The general conditions and neurobehavioral manifestations of the rats were observed;the expression of ubiquitin(UB),ubiquitin activating enzyme(UBE1),ubiquitin conjugating enzyme(UBE2A)and ubiquitin carboxyl terminal hydrolase(UCH-L1)positive cells,protein and mRNA expression levels in the damaged lateral stria-tum of the brain of rats were detected by immunohistochemistry,Western blot and qPCR methods;changes of dopamine(DA)content in the damaged lateral striatum of the brain of rats were detected by ELISA.RESULTS There was no significant difference in general conditions between the normal control group and the sham operation group,and there was no significant difference in the number of ro-tations,suspension time and number of moving grids.Compared to the sham-op group,the model group rats showed poor spirit,irrita-bility,limb tremor,slow movement,dark yellow fur,less food intake,reduced body weight;the modeling increased number of rota-tions,decreased hanging time and number of moving grids(P<0.01),significantly decreased expression of UB,UBE1,UBE2A,and UCH-L1 positive cells,mRNA and protein expression levels in the damaged lateral striatum(P<0.01),and significantly decreased DA content(P<0.01).Compared with the model group,the general condition of the rats in the madopar group and Compound Di-huang Granule groups was improved,the number of rotations was reduced,the hanging time and number of moving grids were increased(P<0.05,P<0.01),the expression of UB,UBE1,UBE2A,and UCH-L1 positive cells,mRNA and protein expression levels were increased(P<0.05,P<0.01),and the DA content was increased(P<0.01);behavioral tests and the expression levels of various in-dicators were most significantly improved in the madopar group and the high-dose Compound Dihuang Granule group(P<0.05,P<0.01),and there was no statistical significance between the two groups.CONCLUSION Compound Dihuang Granule may play a role in treating PD yin-deficiency and dynamic wind syndrome by regulating the expression of ubiquitin-proteasome system-related proteins,reducing abnormal protein aggregation,and thus alleviating DA content deficiency,and the high-dose Compound Dihuang Granule has the best effect.

Objective To observe the effects of compound Dihuang Granules on striatum PINK1,Parkin and α-syn in rats with Parkinson disease yin-deficiency and wind movement syndrome;To explore the mechanism of compound Dihuang Granules in the treatment of Parkinson disease yin-deficiency and wind movement syndrome.Methods 6-hydroxydopamine was injected into the substantia nigra of the midbrain to prepare the model of PD yin-deficiency and wind movement syndrome.Totally 65 model rats were randomly divided into model group,Madopar(150 mg/kg)group,and TCM low-,medium-,and high-dosage(1.75,3.5,7 g/kg)groups.Another 26 rats were divided into normal control group and sham-operation group,with 13 rats in each group.Each administration group was given corresponding medication solution by gavage,the normal control group,sham-operation group and the model group were given equivalent volume of normal saline,once a day for 28 consecutive days.The behavioral changes of rats were detected,ELISA and biochemical assay kits were used to detect ATP content and COX Ⅳactivity in striatum,HE staining was used to observe the morphological changes of the striatum,immunohistochemistry staining,Western blot and RT-qPCR were used to detect the expression of PINK1,Parkin,α-syn protein and mRNA in striatal tissue.Results Compared with the normal control group and the sham-operation group,the rats in the model group showed increased number of rotations and pole climbing time,decreased swimming score(P<0.01),the ATP content and COX Ⅳ activity in striatum tissue decreased(P<0.01),with disordered arrangement and reduced number of neurons,cell swelling,partial nuclear shrinkage,formation of vacuoles,and blurred boundaries,the expressions of PINK1,Parkin protein and mRNA in striatal tissue decreased(P<0.01),and the expressions of α-syn protein and mRNA increased(P<0.01).Compared with the model group,the number of rotations and pole climbing time of rats in each medication group decreased,swimming scores increased(P<0.01),ATP content and COX Ⅳ activity in striatal tissue increased(P<0.05,P<0.01),the number of neurons increased,the arrangement tended to be neat,and the morphology improved,the expressions of PINK1,Parkin protein and mRNA in striatal tissue increased(P<0.05,P<0.01),the expressions of α-syn protein and mRNA decreased(P<0.05,P<0.01).The improvement was most significant in the Medopar group and TCM high-dosage group(P<0.05,P<0.01),and there was no significant difference between the two groups(P>0.05),and TCM groups showed dosage dependence(P<0.05,P<0.01).Conclusion Compound Dihuang Granules can improve the behavioral symptoms of Parkinson disease yin-deficiency and wind movement syndrome,possibly by regulating the expressions of PINK1,Parkin and α-syn in the striatum tissue,thereby alleviating mitochondrial dysfunction,protecting dopaminergic neurons,and playing a role in treating Parkinson disease.

Objective To observe the effect of compound Dihuang Granules on the expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue of Parkinson disease(PD)rats with yin deficiency and wind movement syndrome.Methods 6-Hydroxydopamine was used to inject into the right substantia nigra to prepare PD with yin deficiency and wind movement syndrome rats model.65 rats that successfully modeled were randomly divided into model group,madopar group(madopar 150 mg/kg)and TCM low-,medium-and high-dosage groups(compound Dihuang Granules 1.75,3.5,7 g/kg),with 13 rats in each group;another 13 rats without intervention were selected as the normal control group,and 13 rats were only injected with ascorbic acid solution as sham-operation group.The administration groups were given corresponding drugs by gavage,and the normal control group,sham-operation group and model group were given normal saline by gavage,once a day for 28 consecutive days.The neurobehavioral changes of rats were observed,and the morphology of liver tissue and colonic tissue were observed by HE staining,the protein and mRNA expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue were detected by immunohistochemical staining,Western blot and RT-PCR,respectively.Results Compared with the normal control group and the sham-operation group,the number of rotation circles of rats in the model group increased,with granular degeneration of liver tissue,loose cytoplasm of cells,slight hepatic steatosis,edema of colonic mucosal and submucosal layers,loose connective tissue,infiltration of a small number of lymphocytes,and significant vascular dilation,the expressions of DJ-1,IP3R,GRP75 and VDAC1 protein and mRNA in liver and colonic tissue were significantly decreased(P<0.01).Compared with the model group,the number of rotation circles of rats in madopar group and TCM groups were significantly decreased,with mild granular degeneration of liver cells,loose cytoplasm,orderly arrangement of hepatic cords,mild edema of the colon,a small amount of lymphocyte and granulocyte infiltration,and a small amount of vascular dilation,the protein and mRNA expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver and colonic tissue significantly increased(P<0.05).The TCM high-dosage group showing the most obvious changes,there was no statistical significance with the madopar group(P>0.05).Conclusion Compound Dihuang Granules may restore endoplasmic reticulum-mitochondria homeostasis,improve gut-liver-brain axis mitochondrial function,protect dopamine neurons,and delay PD by regulating the expressions of DJ-1,IP3R,GRP75 and VDAC1 in liver tissue and colonic tissue.

Methcathinone (MCAT) belongs to the designer drugs called synthetic cathinones, which are abused worldwide for recreational purposes. It has strong stimulant effects, including enhanced euphoria, sensation, alertness, and empathy. However, little is known about how MCAT modulates neuronal activity in vivo. Here, we evaluated the effect of MCAT on neuronal activity with a series of functional approaches. C-Fos immunostaining showed that MCAT increased the number of activated neurons by 6-fold, especially in sensory and motor cortices, striatum, and midbrain motor nuclei. In vivo single-unit recording and two-photon Ca²⁺ imaging revealed that a large proportion of neurons increased spiking activity upon MCAT administration. Notably, MCAT induced a strong de-correlation of population activity and increased trial-to-trial reliability, specifically during a natural movie stimulus. It improved the information-processing efficiency by enhancing the single-neuron coding capacity, suggesting a cortical network mechanism of the enhanced perception produced by psychoactive stimulants.
Mice ; Animals ; Reproducibility of Results ; Neurons ; Sensation ; Perception

Objective:To assess the combination of conventional ultrasound with contrast-enhanced ultrasound(CEUS)in the diagnosis of urothelial carcinoma(UC)of the renal pelvis in elderly patients.Methods:Sixty-seven elderly patients with a histopathologic diagnosis of UC of the renal pelvis and surgically treated at our hospital between April 2015 and March 2023 were retrospectively analyzed.Characteristics of regular preoperative 2D ultrasound, color Doppler flow imaging(CDFI)and CEUS were examined.Results:Of 67 patients, 49(73.13%)were found to have localized lesions in the renal pelvis and renal calyces.Lesions in 53 patients(79.10%)could be clearly identified by conventional ultrasound, with 46(86.79%)being isoechoic or hypoechoic, and 7(13.21%)being hyperechoic.Analysis of tumor blood flow by CDFI found 22 cases(41.51%)with avascular lesions, 21(39.62%)with hypovascular lesions and 10(18.87%)with hypervascular lesions.The average value of the resistance index(RI)was 0.64.Enhancement was seen in 62 lesions(92.54%)by CEUS after injection of SonoVue.Compared with the cortex of the ipsilateral kidney, a slow enhancement pattern was observed in 46(74.19%), 14(22.58%)showed simultaneous enhancement, and 2(3.23%)showed fast enhancement.At peak enhancement, 43 lesions(69.35%)had hypo-enhancement, 10(16.13%)had iso-enhancement, and 9(14.52%)had hyper-enhancement, compared with the cortex.Concerning the homogeneity of enhancement, 16(25.81%)displayed heterogeneous enhancement, with tumor necrosis or hemorrhage, and 46(74.19%)had homogeneous enhancement.When the contrast agent washout rate was assessed, a fast washout pattern was observed in 53(85.48%), synchronous washout in 6(9.68%), and slow washout in 3(4.84%).Conclusions:UC of the renal pelvis mostly shows isoechoic and hypoechoic lesions on conventional ultrasound, avascular or hypo-vascular lesions on CDFI, and slow-in, fast-out and hypo-enhancement on CEUS, compared with the cortex.Conventional ultrasound combined with CEUS can help improve the diagnostic accuracy of UC of the renal pelvis.

Purpose: Cavernous nerve injury induced erectile dysfunction (ED) is a refractory complication with high incidence in person under radical prostatectomy. Studies have shown that relaxin-2 (RLX-2) plays a vital role of endothelial protection, vasodilation, anti-fibrosis and neuroprotection in a variety of diseases. However, whether penile cavernous erection can benefit from RLX-2 remains unknown. The purpose of the experiment was to explore the effects of RLX-2 on ED in the rat suffering with bilateral cavernous nerve injury (BCNI). Materials and Methods: The rats were divided into three groups: Sham group was underwent sham operation, BCNI+RLX group or BCNI group was underwent bilateral cavernous nerve crush and then randomly treated with RLX-2 (0.4 mg/kg/d) or saline by continuous administration using a subcutaneously implanted micro pump for 4 weeks respectively. Then, erectile function was evaluated by electrical stimulation of cavernous nerves. Cavernous nerves and penile tissues and were collected for histological evaluation. Results: Erectile function of rats with BCNI was partially improved after RLX-2 treatment. The BCNI group had lower expression of relaxin family peptide receptor (RXFP) 1, p-AKT/AKT, p-eNOS/eNOS ratios than sham operation rats, but RLX-2 could partially reversed these changes. Histologically, the BCNI+RLX group had a significant effect on preservation of neurofilament, neuronal glial antigen 2 of penile tissue and nNOS of cavernous nerves when compared with BCNI group. RLX-2 could inhibited the lever of BCNI induced corporal fibrosis and apoptosis via regulating TGFβ1-Smad2/3-CTGF pathway and the expression of Bax/Bcl-2 ratio, caspase3. Conclusions RLX-2 could improve erectile function of BCNI rats by protecting cavernous nerve and endothelial function and suppressing corporal fibrosis and apoptosis via RXFP1 and AKT/eNOS pathway. Our findings may provide a promising treatment for refractory BCNI induced ED.

Objective:To study the value of shear wave elastography (SWE) combined with contrast-enhanced ultrasound (CEUS) in diagnosing the invasiveness of papillary thyroid microcarcinoma (PTMC), and analyze its risk factors.Methods:This study included 200 patients with pathologically confirmed PTMC who underwent surgery in Fenyang Hospital from January 2019 and June 2021. All were diagnosed with SWE and CEUS before surgery. The value of the two methods in diagnosing the invasiveness of PTMC was explored. The patients′ data were collected to screen the risk factors for the invasiveness of PTMC.Results:It was pathologically confirmed that among the 200 patients with PTMC, there were 112 cases with malignant nodules, 88 cases with benign nodules, 75 cases with cervical lymph node metastasis (including 71 cases with capsular invasion) and 125 cases without lymph node metastasis. CEUS parameters of malignant nodules were significantly higher than those of benign nodules ( P<0.05). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of SWE combined with CEUS to diagnose capsular invasion were 94.66%, 85.60%, 89.00%, 79.77% and 96.39%, with high consistency with the pathological results ( Kappa>0.75). Multivariate Logistic regression analysis showed that multiple foci, irregular shape, breakthrough capsule and small calcification were independent risk factors for the invasiveness of PTMC (VIF<3). The ROC curve results showed that the AUC of SWE combined with CEUS to diagnose capsular invasion was 0.772, and the diagnostic sensitivity and specificity were 73.91% and 80.56%. Conclusions:SWE combined with CEUS can significantly improve the diagnostic accuracy for the invasiveness of PTMC.

Objective:To conduct periodic revalidation of the 15 items and 43 terms autoverification rules of blood analysis after 1 year of application, analyze the application suitability and make the rules improved.Methods:Track the results of 528 010 blood analysis samples of our hospital from August 1, 2019 to January 31, 2020, and analyze the pass rate and interception rate of autoverification; 600 specimens in total were selected randomly for microscope examination, including 300 specimens which touched autoverification rules (1 012 items of autoverification rules) and were intercepted by autoverification and 300 specimens which untouched autoverification rules and were released by autoverification. The abnormal characteristics and unacceptable Delta check of the specimens also need to be concerned at the same time.The false negative rate and false positive rate, true negative rate, true positive rate and pass correct rate of autoverification were verified and compared with the rate of the second phase verification when the autoverification rule was established. The false negative rate, false positive rate, true negative rate and true positive rate of the Delta check rule which 54 716 specimens touched were calculated and compared with the second phase verification rate when the autoverification rule was established.The results of microscopic examination were used as the gold standard for the calculation of the rates, and P<0.05 was considered as a significant difference. The false positive and true positive of 1 012 autoverification rules were analyzed item by item.The false positive and true positive of 108 specimens which touched blast cell autoverification rule were analyzed terms by terms. The mean TAT and median TAT of 528 010 specimens and 193 750 outpatient specimens were calculated respectively, and the report percentages of 528 010 samples that TAT<30, 30-60　and>60 min were calculated respectively. Analyze and evaluate the application suitability of autoverification rules to juge whether they meet the needs of doctors and laboratory. The design process and the rules and application process of autoverification were optimized and improved.Results:The autoverification pass rate was 63.06% (332 971/528 010), the interception rate was 36.94% (195 039/528 010). The false negative rate was 1.00% (1/600), the false positive rate was 12.67% (76/600), the true negative rate was 49% (294/600), the true positive rate was 37.33% (224/600), and the correct rate was 98% (294/300). The pass rate, true negative rate, true positive rate and correct rate of the periodic reverification group were higher than the second phase verification group, the false negative rate and false positive rate were lower than that the second phase verification group. The false negative rate and true positive rate of the Delta check of periodic verification group were lower than that the second phase verification group, the false positive rate and true negative rate were higher than the second phase verification group, there were significant differences in the comparition results. The mean TAT of 528 010 specimens was25 min, and the median TAT was 22 min. The mean TAT of 193 750 outpatient specimens was 23 min, and the median TAT was 20 min. The report percentages of 528 010 samples that TAT<30 min, 30 min-60 min and>60 min were 83.30% (439 819/528 010), 8.00% (42 250/528 010) and 8.70% (45 941/528 010), respectively.Conclusion:The results of periodic revalidation of autoverification after 1 years application show that the 15 items and 43 terms autoverification rules of blood analysis could meet requirements about the accuracy and efficiency of the laboratory, and have a good suitability for application.

Objective To evaluate the effect of double-leaf perforator free flap pedicled with posterolateral calf peroneal artery on repairing facial through-and-through defect after oral cancer oblation.Methods Nine patients with facial through-and-through defects after oblation of oral and maxillofacial malignancies,including 4 cases of buccal mucosa carcinoma,2 cases of maxilla osteosarcoma,1 case of carcinoma of the maxillary sinus and 2 cases of parotid duct carcinoma,were recruited from May,2016 to May,2018.The flap was used to repair the facial defect of recruited patients.The area of the bigger leaf of bilobed perforator flap was 7.0 cm×8.0 cm-6.0 cm×7.0 cm and the small leaf was 4.5 cm×4.5 cm-4.0 cm×4.0 cm,respectively.The intraoral and facial defects needed to be reconstructed contained the oral mucosal of the upper palate,the skin,subcutaneous tissue and mucosa of the cheek.The radiotherapy was performed 1.0-1.5 months after the operation.The prognosis including appearance,mouth opening,and the functions of deglutition,and language were assessed.Results Through 3-18 months outpatient followed-up,all 9 cases of transplanted flaps survived well and the incisions of the donor and recipient areas healed by first intention.The patients were satisfied with the facial appearance,mouth opening,and the functions of deglutition and language at the 6 months follow-up.The radiotherapy had no damage on the survival of the bilobed perforator flap.There was no recurrence or metastasis in follow-up.Conclusion The double-leaf perforator free flap pedicled with posterolateral calf peroneal artery is an ideal free tissue for repairing the facial perforating defect after oral cancer oblation because of its constant blood vessel,abundant tissues,flexible design and operating methods.

Objectives To assess the clinical outcomes,the technical characteristics of percutaneus endoscopic interlaminar decompression for lumbar lateral recess stenosis in elderly patients.Methods From May 2014 to June 2015,76 elderly patients with lumber lateral recess stenosis were accepted percutaneus endoscopic interlaminar decompression,including 48 males and 28 females aged from 60-91 years old (average,71.7:8.8 years).There were 40 patients with stenosis in L45,29 patients with stenosis in L5S1 and 7 patients with stenosis in L4.5 and L5S1.Back and leg pain were assessed by visual analogue scale (VAS).The influence of pain was evaluated by Oswestry disability index (ODI).The change of spinal canal was assessed by lateral recess angle.The MacNab criteria was evaluated at last follow-up.Results All of the 76 operations had been completed successfully with an average operation time of 64.3±23.9 min (range,56-98 min).An average follow-up time was 31.5 months (range,24-36months).There was no operative segment errors and nerve root tear during operation.Dural sac tear in 1 case,2 cases of nerve root injury and 1 case of postoperative kakesthesia,transient seizures after anesthesia occurred in 1 case and 1 case had postoperative delirium.There were significances in lumbocrural pain scores,leg pain scores and ODI scores between preoperation and postoperative 1,3,6,12 month and last follow-up (P＜0.05).In the MacNab last follow-up,the fineness rate was 92.1％,including 52 cases of excellence,18 cases of good,and 6 cases of fair.The latera recess angle between preoperation and postoperation was significantly different.During follow-up period,there were 3 cases developed into lumbar instability.No recurrence,discitis and leakage of cerebrospinal fluid were found,delayed incision healing was found in 1 case.Conclusions Percutaneus endoscopic interlaminar decompression for lumbar lateral recess stenosis in elderly patients is a safe,effective and minimally invasive method of operation with the characteristics of precise decompression of bone stenosis,sufficient resection of ligamentum flavum to decompress soft stenosis,small effect on spinal stability and low complications.