Abstract:Aim To study the effect of rotenone on α-synuclein in rat midbrain of Parkinson's disease(PD).Methods Rats were subcutaneouly injected with chronic low dose rotenone(1.0 mg·kg~(-1)·d~(-1)).Movements within 5 minutes were evaluated in open field test.Tyrosine hydroxylase(TH)-positive neuronsin the midbrain were measured with immunofluorescence staining.α-SYN protein level in the midbrain was demonstrated with immunohistochemical staining and Western blot.Results Compared to the control group,latency time,crossing,rearing and rearing time were changed significantly(P<0.05,0.01)in the rotenone group,TH-positive neurons were reduced(P<0.05)and α-SYN protein level in the midbrain was increased(P<0.05).Conclusion Injection with rotenone can induce PD symptom,which may be correlated to α-SYN protein level in the midbrain.

Objective:To study the anti-inflammatory and analgesic effects of the volatile oil from Piper puberulum ( Benth. ) Max-im. in Guizhou. Methods:Fifty KM mice were randomly divided into five groups (10 ones in each), the model control group (sa-line), low concentration volatile oil group (0. 125 ml/100 ml), medium concentration volatile oil group (0. 25 ml/100ml), high con-centration volatile oil group (0. 5 ml/100 ml) and aspirin group (3 mg·ml-1). Mouse ear swelling induced by xylene and skin capil-lary permeability were used to evaluate anti-inflammatory activity of the volatile oil from Piper puberulum ( Benth. ) Maxim. in Guizhou. Acetic acid-induced abdominal eonstrietion test and hot-plate test were applied to evaluate anti-analgesic activity of the vola-tile oil from Piper puberulum ( Benth. ) Maxim. In Guizhou. Results:Compared with that in the model group, the mouse ear swelling in the medium concentration volatile oil group, high concentration volatile oil group and aspirin group was significantly reduced ( P<0. 05). The skin capillary permeability in the high concentration volatile oil group and aspirin group was significantly reduced (P<0. 05 or P<0. 01). Compared with that in the model group, the writhing times in the medium concentration volatile oil group, high concentration volatile oil group and aspirin group were significantly reduced (P<0. 05 or P<0. 01). The pain threshold in the high concentration volatile oil group was effectively reduced (P<0. 05 or P<0. 01). Conclusion: The volatile oil from Piper puberulum ( Benth. ) Maxim. in Guizhou has promising anti-inflammatory and analgesic activities.

This study is to investigate the amelioration effect of glucocorticoid receptor (GR) antagonist mifepristone on the changes of learning and memory abilities in rat model of depression. In the present study, a 35-day rat chronic unpredictable stress (CUS) model was used to observe both depression-like behaviors with sucrose preference test and open-field test and learning and memory-associated behaviors with Morris water maze test. A total of 45 male adult Sprague-Dawley rats were randomly assigned to three groups of equal size: control group (CON); CUS group (CUS); CUS + mifepristone group (CM). Animals in CM group were first exposed to CUS for 14 days, and then were administered with 50 mg x kg(-1) x d(-1) of mifepristone with continued CUS procedure. Corticosterone EIA Kit was used to detect the concentration of plasma corticosterone (CORT). Nissl staining was used to observe the structure of hippocampus. The results demonstrated that CUS exposure induced both depressive-like and learning and memory-associated behaviors and these deficits were reversed by mifepristone. Compared to CON group, the concentration of plasma CORT increased significantly in CUS group. CUS exposure damaged the structure of hippocampus, whereas mifepristone had an amelioration effect. Together, the structural deficits of hippocampus resulting from long-term stress exposure, which could contribute to the impairment of learning and memory in depression, are reversed by the GR receptor antagonist mifepristone.

Objective To explore optimal initial the best screening time for newborns with different delivery methods using AABR.Methods A total of 550 newborns who were born from August 1, 2016 to October 31, 2016 at our hospital participated in the study.AABR was used to accomplish the initial hearing screening.The newborns were divided into 2 groups according to the delivery methods.There were each 100 neonates born in vaginal during <24 h, 24~48 h and 48~72 h after birth, respectively.The numbers of neonates delivered by cesarean section during the 3 separate periods were 50, 100 and 100, respectively.The newborns who failed the preliminary hearing screening proceeded to the re-screening and diagnostic procedures.Results There were 300 newborns were born in vaginal, and the pass rate in 24~48 h after birth group was significantly higher than that in 24 h group (93.00% vs 83.00%,x2=4.735,P=0.03<0.05), but it was not significantly different from that of 48~72 h group (95.00% vs 93.00%,x2=0.355,P=0.56>0.05).There were 250 newborns in cesarean section, the pass rate of 24~48 h after birth group was significantly higher than that in 24 h group (83.00% vs 68.00%,x2=4.437, P=0.04<0.05), and significantly lower than that of 48~72 h group (94.00% vs 83.00%,x2=5.944, P=0.02<0.05).Conclusion Taking into account of hospitalization time, the screening time for the vaginal delivery newborn hearing screening can be advanced to 24~48 h after birth with the application of AABR, but not for the cesarean section group.

Objective To observe the effect of Huoxue Jiedu Recipe(Herbal recipe for activating blood and removing toxins)on proliferation and cell cycle status of keratinocytes cultured in vitro in psoriasis.Methods Using the serum pharmacologic technique,the clean SD rats were adopted to make the Huoxue Jiedu Recipe medicated serum(high,moderate,low dose),Dexamethasone medicated serum,and blank serum,which were added into the keratinocytes culture system respectively.After incubated for 48 hours,the cell proliferation of keratinocytes was measured by MTT assay,and the cell cycle distribution estimated with flow cytometer(FCM).Results The different density of Huoxue Jiedu Recipe could all inhibit significantly the generation of COLO-16 keratinocyte line and change the distribution proportion of each phase in the cell division cycle.Conclusion Huoxue Jiedu Recipe treats psoriasis through inhibiting the proliferation of keratinocytes.

Aim To construct the cell model targeted on the damage by α-synuclein for screening anti-Parkinson’s Disease (PD)compounds.Methods The cDNA fragment of α-synucle-in gene was obtained by PCR methods and inserted into the re-combinant prokaryotic plasmid by molecular cloning technique. The recombinant plasmid was transformed into Escherichia coli, and subsequently induced to express α-synuclein protein.The recombinant α-synuclein was purified and identified by affinity chromatography,immunoblotting and mass spectrometry.The cells damage by α-synuclein was evaluated through cell viability measured by 3-(4,5-dimethyl-2-thiazolyl )-2,5-diphenyl-2-H-tetrazolium bromide.Results The obtained cDNA fragment ofα-synuclein in accordance with its theoretic molecular weight was cloned into pET30a plasmid and verified by sequencing.The re-combinant plasmid was transformed into bacteria E.Coli.BL21 (DE3)and induced to express α-synuclein by isopropyl β-D-1 -thiogalactopyranoside (IPTG).The expression condition was op-timized according to the culture temperature,the concentration of IPTG and the proliferation state of bacteria.The purified α-synu-clein was proved to be a 1 5.3 ku molecule weight protein,and could be immunoblotted with anti-α-synuclein antibody.The pu-rified α-synuclein could decrease the viability of PC1 2 cells and primary neurons significantly,and its effect was in a concentra-tion-dependent manner.Conclusion We have succeeded in constructing the cell model targeted on the damage by α-synucle-in.

Aim To study of the expression and distribution of four α-synuclein truncations in three cells.Method Four α-synuclein gene truncations were obtained by PCR method,followed by subcloning into the pEGFP-N1 eukaryotic expression vector.Four obtained recombination plasmids were transfected into MN9D cells,PC12 cells and SH-SY5Y cells using Lipofectamine 2000 respectively.The expression and distribution of four α-synuclein truncations were observed by Confocal.Results Distribution of four α-synuclein truncations was discrepant obviously,the truncations,with more C terminal remained,were prone to emerging in nuclei.Conclusion Localization of α-synuclein protein in cells may be related to the C terminal,and the whole C terminal plays an important role in distribution of α-synuclein into nuclei.

Objective:To analyze the clinical features and genetic background of adrenoleukodystrophy(ALD).Methods:In this study, we reported a rare case of ALD who initially presented with progressive bilateral lower limb weakness. The clinical data of the patient and his family members were collected and the ABCD1 gene was sequenced for the patient and his three daughters by a high-throughput sequencing method.Results:The proband had a later onset of symptoms, a prolonged course of the disease, and initially exhibited bilateral lower limb weakness and nocturnal muscle spasms. The disease progressed to spastic quadriplegia, aphasia, dementia, swallowing difficulties, and urinary and fecal incontinence. Serum very-long-chain fatty acid concentrations were elevated. Subclinical cortisol secretion abnormalities were observed. Cranial imaging indicated symmetrical reduction in density around the lateral ventricles and white matter degeneration. The proband′s ABCD1 gene analysis revealed a novel heterozygous mutation c. 1367G>A, p. R456H. His three daughters carried the same nucleotide heterozygous mutation.Conclusion:This study investigates the clinical characteristics of ALD, providing additional clinical evidence for the diagnosis and treatment of this condition. Additionally, a novel mutation in the ABCD1 gene was identified, contributing to the genetic variation database.

Background: To enhance perioperative outcomes, a perioperative registry that integrates high-quality real-world data throughout the perioperative period is essential. Singapore General Hospital established the Perioperative and Anesthesia Subject Area Registry (PASAR) to unify data from the preoperative, intraoperative, and postoperative stages. This study presents the methodology employed to create this database. Methods: Since 2016, data from surgical patients have been collected from the hospital electronic medical record systems, de-identified, and stored securely in compliance with privacy and data protection laws. As a representative sample, data from initiation in 2016 to December 2022 were collected. Results: As of December 2022, PASAR data comprise 26 tables, encompassing 153,312 patient admissions and 168,977 operation sessions. For this period, the median age of the patients was 60.0 years, sex distribution was balanced, and the majority were Chinese. Hypertension and cardiovascular comorbidities were also prevalent. Information including operation type and time, intensive care unit (ICU) length of stay, and 30-day and 1-year mortality rates were collected. Emergency surgeries resulted in longer ICU stays, but shorter operation times than elective surgeries. Conclusions The PASAR provides a comprehensive and automated approach to gathering high-quality perioperative patient data.

Objective:To establish a method for rapid detection of OXA and par C resistance genes of Acinetobacter baumannii(Ab)by double nucleic acid colloidal gold strip and to develop kit.Methods:DNA of Ab was extracted by heating and boiling method.OXA and par C genes sequences of Ab were selected as target gene fragments based on NCBI.Primers were designed and labeled with 6-FAM,digoxin and biotin,respectively.Drug resistance gene detection reagents were developed,and nucleic acid gold test strips were used for rapid and visual detection.Molecular cloning and sequencing techniques were used to clone positive control samples and evaluate specificity,sensitivity and stability of kit.Results:DNA concentration and purity of Ab extracted by boiling method were good.Homology between cloned and sequenced plasmid DNA and gene sequence in GenBank database was 100%,respectively.Speci-ficity of kit was good,with only Ab showing positive results and other bacterial genera showing negative results;DNA concentration of Ab in double nucleic acid colloidal gold test strip decreased to 10-3 ng/μl,a red line still appeared,whose sensitivity was 10 times higher consistent with minimum detection limit of electrophoresis 10-2 ng/μl;test kits were tested at 3rd,6th and 9th months,and showed good stability.Conclusion:Double resistance detection kit established in this study can simultaneously detect OXA and par C resis-tance of Ab,who has advantages of high sensitivity,strong specificity,rapid and simple,and provides a new method for detection of carbapenem and quinolone antibiotic resistance of Ab.