OBJECTIVE To investigate the variations in taste， aroma and volatile organic compounds of Rhei Radix et Rhizoma decoction pieces derived from different sources， and to identify their origins. METHODS The flavor， odor and volatile organic compounds of Rhei Radix et Rhizoma decoction pieces from different sources were compared and analyzed by using electronic tongue， electronic nose， and gas chromatography-ion mobility spectrometry （GC-IMS）. Principal component analysis （PCA）， partial least squares-discriminant analysis （PLS-DA）， orthogonal partial least squares discriminant analysis （OPLS-DA） and Fisher discriminant analysis were employed to identify the origins of Rhei Radix et Rhizoma decoction pieces and establish the basis discrimination criteria. RESULTS The differences in taste of Rhei Radix et Rhizoma decoction pieces from 3 origins were primarily characterized by bitterness， astringency， and bitter-astringent aftertaste. In terms of smell， variations were mainly observed in inorganic sulfides， organic sulfides containing aromatic components， methane and other short-chain alkanes， alcohols， ethers， aldehydes and ketones， as well as nitrogen oxides. Differentially volatile organic compounds mainly consisted of alcohols， aldehydes and ketones. Furthermore， the samples from 8 batches could be effectively classified into 3 categories.Three types of Rhei Radix et Rhizoma decoction pieces can be effectivily identified based on the peak intensity ratio between volatile substances. For example， when the peak intensity of 2-acetylfuran was 3-19 times that of isobutyric acid [dimer]， it was considered as Rheum officinale Baill. CONCLUSIONS The discriminant models established in this study， along with the criteria for determining the origins based on the peak intensity ofcharacteristic volatile compounds， can be utilized for the identification of Rhei Radix et Rhizoma decoction pieces.

Huntington's disease (HD) is an autosomal dominant inherited disease with insidious onset and slow progression, mainly characterized by chorea-like symptom, intelligence decline, and psychiatric abnormalities. Cause of the disease is abnormal expansion of CAG trinucleotide repeat sequences in the first exon of the Huntington gene (HTT) on chromosome 4. Despite the clear etiology, currently, no effective therapeutic measures to control the disease progress is noted, and symptomatic treatment is still the main treatment in clinical practice. This article provides a brief overview of the current clinical trials, clinical challenges, and future development of HD gene therapy to provide references for subsequent related research.

Lewy bodies formed by abnormal deposition of α-synuclein (α-syn) in neurons and glial cells are diagnostic markers of α-synucleinopathies. However, the early diagnosis of α-synucleinopathies is delayed due to the difficulty of brain biopsy at present. Because that α-syn can produce prion-like replication and spread to the periphery under pathological conditions, the ultramicro detection of pathologic α-syn in extracranial tissues by real-time quaking-induced conversion (RT-QuIC) technique can provide a basis for early diagnosis of α-synucleinopathies. In this paper, we review RT-QuIC technique in detecting α-syn seeding activity in different tissues of patients with α-synucleinopathies to summarize the potential value of RT-QuIC technique in early diagnosis or differential diagnosis of α-synucleinopathies.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.

Objective:To investigate the characteristics and evolution of mild motor symptoms (MMS) in patients with prodromal Parkinson′s disease (pPD).Methods:Based on the pPD cohort screened by Parkinson′s Disease Prodromal Clinical Assessment Scale in Nanjing community from July 2018 to December 2020, the clinical data of 30 patients with pPD who completed the baseline assessment and were followed up for at least 1 year were analyzed. According to the Unified Parkinson Diease Rating Scale Ⅲ (UPDRS-Ⅲ) score, the patients were divided into MMS group (UPDRS-Ⅲ score>3) and non-MMS group (NMMS group, UPDRS-Ⅲ score≤3). The differences and evolution characteristics of clinical characteristics between the 2 groups were compared. Multivariate linear regression was used to analyze the risk factors of motor symptom progression in pPD patients.Results:Among the 30 patients with pPD, 7 of 23 patients in the MMS group were converted to PD at the end of follow-up, 1 of 7 patients in the NMMS group were converted to PD at the end of follow-up. The UPDRS-Ⅲ score [10.00 (7.00, 17.00)], Montreal Cognitive Assessment Scale (MoCA) score [25.50 (24.75, 28.00)] and the Hamilton Anxiety Scale (HAMA) score [9.00 (5.00, 13.00)] at the end of follow-up of pPD patients were significantly higher than those at baseline [7.00 (4.00, 12.00), 24.00 (22.75, 25.25) and 8.00 (2.00, 11.00)], and the differences were statistically significant ( Z=-3.505, P<0.001; Z=-2.956, P=0.003; Z=-2.427, P=0.015).Subgroup analysis showed that UPDRS-Ⅲ score [11.00 (7.00, 18.00)], MoCA score [25.00 (24.00, 27.00)] and HAMA score [ 9.00 (6.00, 15.00)] at the end of follow-up in the MMS group were higher than those at baseline [8.00 (6.00, 12.00), 24.00 (22.00, 25.00) and 9.00 (3.00, 11.00)], and the difference was statistically significant (Z=-2.768, P=0.006; Z=-2.457, P=0.014; Z=-2.250, P=0.024). The Non-Motor Symptoms Questionnaire score at the end of follow-up in the MMS group (8.96±5.20) was significantly lower than that in the baseline (11.04±4.41), and the difference was statistically significant ( t=2.441, P=0.023).There was no significant difference in Mini-Mental State Examination (MMSE), Hamilton Depression Scale (HAMD), Rapid Eyes Movement Sleep Behavior Disorder Questionnaire-Hong Kong (RBDQ-HK) and Sniffin′ sticks olfactory test score at the end of follow-up in the MMS group. Only UPDRS-Ⅲ score in the NMMS group was increased at the end of follow-up [7.00 (5.00, 8.00)] compared with the baseline [4.00 (1.00, 4.00)], and the difference was statistically significant ( Z=-2.375, P=0.018). There was no significant difference in MoCA, MMSE, HAMA, HAMD, RBDQ-HK, and Sniffin′ sticks olfactory test score between the NMMS group and the baseline at the end of follow-up. Conclusion:The clinical conversion rate of pPD patients with MMS is high,and screening of this population should be paid attention.

Background::Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s dementia. Mitochondrial dysfunction is involved in the pathology of PD. Coiled-coil-helix-coiled-coil-helix domain-containing 2 (CHCHD2) was identified as associated with autosomal dominant PD. However, the mechanism of CHCHD2 in PD remains unclear.Methods::Short hairpin RNA (ShRNA)-mediated CHCHD2 knockdown or lentivirus-mediated CHCHD2 overexpression was performed to investigate the impact of CHCHD2 on mitochondrial morphology and function in neuronal tumor cell lines represented with human neuroblastoma (SHSY5Y) and HeLa cells. Blue-native polyacrylamide gel electrophoresis (PAGE) and two-dimensional sodium dodecyl sulfate-PAGE analysis were used to illustrate the role of CHCHD2 in mitochondrial contact site and cristae organizing system (MICOS). Co-immunoprecipitation and immunoblotting were used to address the interaction between CHCHD2 and Mic10. Serotype injection of adeno-associated vector-mediated CHCHD2 and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration were used to examine the influence of CHCHD2 in vivo.Results::We found that the overexpression of CHCHD2 can protect against methyl-4-phenylpyridinium (MPP+)-induced mitochondrial dysfunction and inhibit the loss of dopaminergic neurons in the MPTP-induced mouse model. Furthermore, we identified that CHCHD2 interacted with Mic10, and overexpression of CHCHD2 can protect against MPP +-induced MICOS impairment, while knockdown of CHCHD2 impaired the stability of MICOS. Conclusion::This study indicated that CHCHD2 could interact with Mic10 and maintain the stability of the MICOS complex, which contributes to protecting mitochondrial function in PD.

Objective:To explore the effect of hydrogen sulfide (H 2S) on modulating the subunit Kir6.2 of adenosine triphosphate sensitive potassium channels via the cyclic guanosine monophosphate-dependent protein kinase (cGMP/PKG) signaling pathway in epileptic rat models. Methods:Sixty adult male SD rats were randomly divided into the following six groups (10 rats in each group) by random number table method: control, epileptic, H 2S donor, H 2S donor+epileptic, KT5823 (one inhibitor of the cyclic guanosine monophosphate-dependent protein kinase)+H 2S donor+epileptic, and glibenclamide (one inhibitor of the adenosine triphosphate sensitive potassium channels)+H 2S donor+epileptic groups. Except the control group, SD rats were intraperitoneally injected with plentylenetetrazole to make the kindling models and their behaviours were recorded including the latency period, the grade, and the duration of the first epileptic seizure according to the Racine′s standard. The waveforms of electroencephalogram (EEG) in hippocampus were also recorded during the seizure. The mRNA and protein levels of PKG and Kir6.2 in hippocampus were evaluated by Western blotting and quantitative real-time polymerase chain reaction, and the hippocampal concentrations of cGMP and phosphorylation of cyclic guanosine monophosphate-dependent protein kinase (p-PKG) were detected by enzyme linked immunosorbent assay. Results:Rats in the epileptic group showed Ⅳ-Ⅴ grade of epileptic seizure [4.500 (4.000, 4.875)], short latency period [(10.37±8.21) min] but long duration [(69.50±24.37) s] of seizure. Compared to the epileptic group, rats in the H 2S donor group showed Ⅱ-Ⅲ grade of epileptic seizure ( P=0.004), significantly longer latency period ( P<0.001), and shorter duration of seizure ( P<0.001). Compared to the H 2S donor+epileptic group, rats in the KT5823+H 2S donor+epileptic group showed Ⅲ-Ⅳ grade of epileptic seizures, significantly shorter latency period ( P<0.001), and longer duration of seizure ( P<0.001). The results of EEG showed that the wave patterns in the epileptic group were spike or sharp waves and the amplitudes were largest [(190.570±23.590) μV]. Compared with the epileptic group, amplitudes were reduced ( P<0.001) in the H 2S donor+epileptic group. PKG mRNA and PKG protein were expressed differently among all groups (PKG mRNA: n=5, H=26.714, P<0.001; PKG protein: n=5, F=30.597, P<0.001). Compared with the control group, the expression of both PKG mRNA and PKG protein was decreased (PKG mRNA: 1.000±0.001 vs 0.782±0.064, P=0.023; PKG protein: 0.550±0.037 vs 0.145±0.020, P=0.042) in the epileptic group. Besides, Kir6.2 mRNA and Kir6.2 protein were expressed differently among all groups (Kir6.2 mRNA: n=5, H=27.761, P<0.001; Kir6.2 protein: n=5, F=60.659, P<0.001). Compared with the control group, the expression of both Kir6.2 mRNA and Kir6.2 protein was decreased (Kir6.2 mRNA: 1.000±0.001 vs 0.897±0.033, P=0.004; Kir6.2 protein: 0.384±0.035 vs 0.215±0.016, P=0.024) in the epileptic group. And the concentrations of cGMP and p-PKG were decreased (cGMP: P<0.001; p-PKG: P<0.001) in the epileptic group. The results in the H 2S donor+epileptic group were up-regulated (PKG mRNA: P=0.047; PKG protein: P<0.001; Kir6.2 mRNA: P=0.011; Kir6.2 protein: P<0.001; cGMP: P<0.001; p-PKG: P<0.001) compared with the epileptic group. However, the results in the KT5823+H 2S donor+epileptic group were down-regulated (PKG mRNA: P=0.015; PKG protein: P=0.027; Kir6.2 mRNA: P=0.013; Kir6.2 protein: P=0.017; cGMP: P=0.005; p-PKG: P<0.001) compared with the H 2S donor+epileptic group. Conclusion:A possible mechanism is that H 2S prevents the epileptic seizure from modulating the subunit Kir6.2 of ATP sensitive potassium channels via the cGMP/PKG signaling pathway.

Objective:To evaluate the microstructural changes of brain parenchyma in patients with essential hypertension by diffuse kurtosis imaging (DKI) and enhanced T2 star weighted angiography (ESWAN).Methods:A prospective study was performed; 27 patients with essential hypertension, admitted to our hospital from April 2019 to September 2019, and 16 healthy subjects matched with gender, age and education level were enrolled in our study. According to the presence or absence of cerebral microbleeds (CMBs), patients with essential hypertension were divided into essential hypertension with CMBs group ( n=8) and essential hypertension without CMBs group ( n=19). MRI, DKI and ESWAN were performed on all subjects. The mean kurtosis (MK), mean diffusivity (MD) and fractional anisotropy (FA) of bilateral hippocampal gyrus, centrum semiovale, caudate head, posterior limb of internal capsule, thalamus, red nucleus, substantia nigra, pons, and cerebellum were measured. Results:As compared with the healthy subjects, the patients with essential hypertension had significantly lower MK values in the left semioval center, bilateral caudate head, left posterior limb of internal capsule, and bilateral thalamus, significantly higher MD value in the right thalamus, and statistically lower FA value in the left posterior limb of internal capsule ( P<0.05). The essential hypertension with CMBs group had significantly lower MK values in left hippocampus gyrus, left centrum semiovale, bilateral caudate head, left posterior limb of internal capsule, bilateral thalamus, and left substantia nigra, significantly higher MD value in right thalamus, and significantly lower FA value in left posterior limb of internal capsule as compared with essential hypertension without CMBs group and healthy control group ( P<0.05). Conclusion:In patients with essential hypertension, the brain microstructural changes are found in the hippocampus, centrum semiovale, caudate head, posterior limb of internal capsule, thalamus and substantia nigra; these changes are more obvious in essential hypertension patients with CMBs; DKI and ESWAN can effectively assess the early brain microstructure changes in patients with essential hypertension.

Objective:To investigate the prevalence and clinical characteristics of mild cognitive impairment (MCI) and its related risk factors in patients with prodromal Parkinson's disease (pPD).Methods:Forty-seven pPD patients from Nanjing community and 37 healthy controls (HCs) were recruited in the present study. The pPD patients were divided into pPD-MCI group and normal cognition (pPD-NC) group according to Beijing version of Montreal Cognitive Assessment (BJ-MoCA). The general clinical data, Hamilton Depression Scale (HAMD) scores, Hamilton Anxiety Scale (HAMA) scores, number of patients with rapid eye movement-sleep behavior disorder (RBD), overall cognitive function and cognitive domain impairment of members from pPD group and HCs, pPD-MCI group and pPD-NC group were respectively compared. Risk factors for pPD-MCI were analyzed by multivariate Logistic regression analysis.Results:The prevalence of pPD-MCI in the 47 pPD patients was 57.45% (27/47). As compared with the HCs, pPD patients had significantly higher HAMD scores and HAMA scores, statistically larger number of patients with RBD, while significantly lower overall Montreal Cognitive Assessment (MoCA) scores and statistically lower scores in cognition domains of visuospatial and executive function, attention, abstraction, delayed memory and orientation ( P<0.05). Patients in pPD-MCI group had significantly higher HAMA scores, Unified Parkinson's Disease Rating Scale-Motor Examination scores, statistically lower education level, significantly lower MoCA scores, and statistically lower scores in cognition domains of visuospatial and executive function, abstraction, and delayed memory as compared with patients in pPD-NC group ( P<0.05). Multivariate Logistic regression analysis showed that low education level was an independent risk factor for pPD-MCI ( OR=0.800, 95%CI: 0.650-0.985, P=0.035). Conclusions:The prevalence of MCI in pPD patients is high, and multiple cognitive domains can be impaired in early stage. Patients with pPD-MCI are more vulnerable to depression and mild motor symptoms, so attention should be paid to emotional intervention and sport life guidance for patients at early stage. Education level is crucial for pPD-MCI, and cognitive training may contribute to slow down the process of MCI.

Objective:To investigate the characteristics of senile neurodegenerative diseases (NDDs) inpatients in south China, especially in Guangdong province, and explore the influencing factors for their medical expenses.Methods:The medical records of 7231 patients with NDDs≥65 years were collected in the electronic health database of our hospital from January 2010 to December 2019, including gender, age, admission ways, chief complaints, length of hospital stays and medical expenses. On the basis of median of the medical expenses (21 345 yuan) of these patients, they were divided into low cost (<21 345 yuan) group and high cost (≥21 345 yuan) group. Univariate Logistic analysis and multivariate Logistic regression analysis were conducted to screen the influencing factors for medical expenses and the independent influencing factors.Results:(1) The main age group of geriatric inpatients with NDDs were 70-79 years (40.96%); the admission source was mainly outpatient (56.70%), and length of hospital stays of a large percent of patients (44.50%) were 8-14 d. (2) From 2010 to 2019, the number of hospitalized geriatric patients with NDDs showed an increasing trend year by year, the overall trend of length of hospital stays was shortened, and the medical expenses showed gradual increase; the causes of hospitalization, percentages of patients caused by infection, abnormal blood pressure and water-electrolyte metabolism disturbances showed decreased trend, percentages of patients caused by heart diseases, cerebrovascular accidents and mental-psychological diseases showed increased trend, and the proportions of patients caused by fracture/trauma/wound injuries were generally stable. The proportion of patients returning home and mortality rate after hospital discharge were declined, and the proportion of patients returning to other medical or community institutions was increased. (3) Living in ICU, length of hospital stays, diabetes, nosocomial infection, chronic kidney disease, urinary tract infection, tumble, body mass index, and anticholinergic drugs were independent risk factors influencing the medical expenses ( P<0.05). Conclusions:An aging trend is noted in patients with NDDs; the number of hospitalized patients and medical expenses increase year by year, and the length of hospital stays gradually decreases. In view of the many factors that influence the medical expenses of this disease, it is suggested to develop the corresponding standardized treatment plan for the main influencing factors in clinical practice.