[Analysis of changes in intrinsic neural timescales in male smoking addicts based on whole brain resting-state functional magnetic resonance imaging].

Objective: To investigate the abnormal changes of intrinsic neural time scale (INT) in male smoking addicts based on whole brain resting state functional magnetic resonance imaging (rs-fMRI). Methods: A case-control study. The clinical data and whole brain rs-fMRI data of 139 male subjects, aged (34.1±8.8) years, recruited through the online platform from January 2019 to December 2021 were retrospectively analyzed. According to the existence of smoking addiction, they were divided into smoking addiction group (n=83) and healthy control group (n=56).INT was calculated to reflect the brain neural activity dynamics. Single sample t test was used to obtain the whole brain spatial distribution maps of INT in smoking addiction group and the control group. Then two-sample t test was conducted to explore the difference of INT between the smoking addition group and the healthy control group, with age and years of education as covariates. Finally, Spearman correlation analysis was used to explore the relationship between INT and nicotine dependence scale score and smoking index. Results: Subjects with smoking addiction and healthy control group showed a similar pattern of hierarchical neural timescales, namely shorter INT in sensorimotor areas and longer INT in parietal lobe, posterior cingulate cortex. In addition, in the smoking addiction group, the left medial occipital gyrus (peak t=-3.18), left suproccipital gyrus (peak t=-3.66), bilateral pericalar cleft cortex (left: peak t=-3.02, right: peak t=-3.22), bilateral lingual gyrus (left: peak t=-3.10, right: t peak=-3.04), left cuneus (peak t=-2.97), default network associated brain region [left anterior cuneus(peak t=-3.23), left angular gyrus (peak t=-3.07), and left posterior cingulate cortex (peak t=-3.54) were significantly lower than those of healthy controls (gaussian random field correction, voxel level all P<0.005, mass level all P<0.05). However, there was no significant correlation between INT and nicotine dependence scale score and smoking index (both P>0.05 after Bonferroni correction). Conclusion: Compared with healthy controls, smoking addicts showed abnormal changes in the dynamics of neural activity in the visual cortex and the default network.

[Genetics and clinical phenotypes of epilepsy associated with chromosome 2q24.3 microdeletion].

Objective: To summarize the genetics and clinical phenotypes of epilepsy children with 2q24.3 microdeletion. Methods: All the patients with 2q24.3 microdeletion were retrospectively collected at the Pediatric Department of Peking University First Hospital from March 2017 to July 2022. The features of clinical manifestations, electroencephalogram (EEG), and neuroimaging were analyzed. Results: There were 13 patients with 2q24.3 microdeletion were included. All 13 patients had de novo copy number variation (CNV) with a deletion size ranged 0.18-7.31 Mb. The main pathogenic genes in the region were SCN3A, SCN2A, TTC21B, SCN1A and SCN9A genes. Among the 13 patients, 7 were boys, and 6 were girls. The onset age of epilepsy was 3.3(2.5, 6.0) months. Multiple seizure types were observed, including focal seizures in 13 patients, generalized tonic-clonic seizures (GTCS) in 6 patients, myoclonic seizures in 3 patients, epileptic spasm in 2 patients, and tonic seizures in 2 patients. Seizures were fever sensitivity in 9 patients. Status epilepticus was observed in 6 patients. One case had normal mental motor development and 12 cases had different degrees of developmental delay. Six patients had craniofacial abnormality, 1 had six-finger deformity of the right thumb, and 1 had multiple system abnormalities. EEG showed focal discharge in 3 cases, multifocal discharges in 5 cases, multifocal and generalized discharges in 1 case. Brain magnetic resonance imaging (MRI) showed enlargement of subarachnoid spaces in the frontal and temporal region in 4 patients, enlargement of lateral ventricle in 4 patients and delayed myelination of white matter in 1 patient. Dravet syndrome was diagnosed in 5 cases. The age at the last follow-up were 2.5(1.4,5.5) years, 1 patient was seizure free longer than 1 year, and 12 patients still had seizures. Conclusions: The epilepsy associated with 2q24.3 microdeletion is mainly induced by the deletion of SCN3A, SCN2A and SCN1A genes. The seizure onset age of 2q24.3 microdeletion related epilepsy was in infancy. Multiple seizure types are observed and the common seizure types include focal seizures and GTCS. Most patients have fever sensitivity and status epilepticus. Most patients have developmental delay. The phenotype of patients with deletion of SCN3A and SCN2A gene is more severe than that of patients with deletion of SCN1A gene only.

[Genotypes and phenotypes of IQSEC2 gene variants related epilepsy].

Objective: To analyze the genotypes and clinical phenotypes of patients with epilepsy associated with IQSEC2 gene variants. Methods: The genotypes, seizure types, electroencephalogram, neuroimage of 6 patients with IQSEC2 gene variants in the Department of Pediatrics, Peking University First Hospital from July 2019 to October 2021 were analyzed. Results: There were 5 males and 1 female. Six variants were de novo, including 2 frameshift variants (c.3801_3808dup/p.Q1270Rfs*130, c.1459_1460delAT/p.M487Vfs*2), 2 nonsense variants (c.3163C>T/p.R1055*, c.1417G>T/p.E473*), 1 in-frame deletion (c.2295_2297del/p.N765del) and 1 missense variant (c.2293A>G/p.N765D). Age at seizure onset ranged from 3 months to 2 years and 5 months. Multiple seizure types were observed, including epileptic spasms in 5 patients, focal seizures in 5 patients, tonic seizures in 3 patients, myoclonic seizures in 3 patients, atypical absence seizures in 2 patients and atonic seizures in 2 patients. All 6 patients showed global developmental delay before seizure onset. There were other clinical manifestations, including autistic features in 3 patients, microcephaly in 3 patients, dystonia in 2 patients and binocular esotropia in 1 patient. The electroencephalogram showed slow background activity and hypsarrhythmia in all 6 patients. Brain magnetic resonance imaging showed abnormal in 5 patients and normal in 1 patient. Five patients were diagnosed with infantile spasms. Among them, 4 patients had late-onset infantile spasms. One patient was unclassified developmental epileptic encephalopathy. The age of last follow-up ranged from 3 years and 2 months to 7 years and 2 months. All 6 patients still had seizures after multiple anti-seizure medications. Conclusions: The seizure onset of patients with IQSEC2 gene variants usually begins after 1 year of age. The common seizure types include epileptic spasms and focal seizures. Patients usually have a global developmental delay before seizure onset. IQSEC2 variants could be related to developmental and epileptic encephalopathy, and most patients are diagnosed with late onset infantile spasms. Epilepsy associated with IQSEC2 gene variants is usually refractory.

[Clinical phenotypes and genetic features of epilepsy children with MBD5 gene variants].

Objective: To summarize the phenotypes of epilepsy in patients with MBD5 gene variants. Methods: A total of 9 epileptic patients, who were treated in the Department of Pediatrics, Peking University First Hospital from July 2016 to September 2021 and detected with MBD5 gene pathogenic variants, were enrolled. The features of clinical manifestations, electroencephalogram (EEG), and neuroimaging were analyzed retrospectively. Results: Among 9 patients, 6 were male and 3 were female. Age at seizure onset ranged from 5 to 89 months. Multiple seizure types were observed, including generalized tonic clonic seizures (GTCS) in 7 patients, myoclonic seizures in 5 patients, focal seizures in 5 patients, atypical absence seizures in 3 patients, atonic seizures in 2 patients, myoclonus absence seizures in 1 patient, epileptic spasms in 1 patient, and tonic seizures in 1 patient. There were 8 patients with multiple seizure types, 2 patients with sensitivity to fever and 5 patients with clustering of seizures. Two patients had a history of status epilepticus. All patients had developmental delay before seizure onset. Nine patients had obvious language delay, and 6 patients had autism-like manifestations. Five patients had slow background activity in EEG. Interictal EEG showed abnormal discharges in 9 patients. Brain magnetic resonance imaging (MRI) was normal in all patients. A total of 9 epileptic patients carried MBD5 gene variants, all of them were de novo variants. There were MBD5 gene overall heterozygous deletion in 1 patient, large fragment deletions including MBD5 gene in 3 patients and single nucleotide variations (c.300C>A/p.C100X, c.1775delA/p.N592Tfs*29, c.1759C>T/p.Q587X, c.150_151del/p.Lys51Asnfs*6, c.113+1G>C) in 5 patients. The age at last follow-up ranged from 2 years and 9 months to 11 years and 11 months. At the last follow-up, 2 patients were seizure-free for more than 11 months to 4 years 6 months, 7 patients still had seizures. Conclusions: The initial seizure onset in patients with MBD5 gene variants usually occurs in infancy. Most patients have multiple seizure types. The seizures may be fever sensitive and clustered. Developmental delays, language impairments, and autistic behaviors are common. MBD5 gene variants include single nucleotide variations and fragment deletions. Epilepsy associated with MBD5 gene variants is usually refractory.

[Genotypes and clinical features of neonatal-onset genetic epilepsy in 141 patients].

Objective: To summarize the genotypes and clinical features of neonatal-onset genetic epilepsy. Methods: Patients (114 cases) with identified gene variants were collected from May 2013 to May 2019 in Peking University First Hospital, retrospectively. The genotype, clinical, electroencephalographic and neuroimaging characteristics were analyzed. Results: A total of 141 neonatal-onset epilepsy patients with identified gene variants were enrolled, including 76 males and 65 females and involving 33 epilepsy genes. Top five genes were KCNQ2 (56 cases), SCN2A (25 cases), STXBP1 (9 cases), CDKL5 (8 cases) and KCNT1 (6 cases), accounting for 73.8% (104/141). The age of seizure onset was 3(1-28) days of age, 71.6% (101/141) were within 1 week of age. The age of genetic diagnosis was 4 months (1 month to 13 years) of age. A total of 130 patients presented focal seizures; 47 patients presented epileptic spasms. Other seizure types included generalized tonic-clonic seizures, clonic seizures, myoclonic seizures, tonic seizures and absence seizures. Fifty-eight patients experienced multiple seizure types. The results of video-electroencephlogram (VEEG) were abnormal in 127 patients and in 62 patients clinical seizures were captured. Global developmental delay was presented in 122 patients. Epilepsy syndromes were diagnosed in 59 patients. Thirteen patients were diagnosed as Ohtahara syndrome (OS), 9 as epilepsy of infancy with migrating focal seizures (EIMFS), 17 as West syndrome (WS), 4 as OS developed to WS, 9 as benign neonatal epilepsy (BNE), 2 as benign familiar neonatal-infantile epilepsy (BFNIE), 2 as benign infantile epilepsy (BIE) and 3 as benign familial infantile epilepsy (BFIE). Sixty-seven patients were diagnosed as unclassified early infantile epileptic encephalopathy (EIEE), 13 patients could not be diagnosed as any epilepsy syndrome, and 2 patients were diagnosed as pyridoxine-dependent epilepsy. Forty-six patients had abnormal neuroimaging including cortical atrophy, corpus callosum dysplasia and cerebellar atrophy, involving 19 genes. Conclusions: Neonatal-onset epilepsy is related to many different genes. Seizure onset age of most patients is within one week after birth. Focal seizures and epileptic spasms are more common. Some patients show abnormal neuroimaging.

[Genotype and phenotype of children with DEPDC5 gene variants related epilepsy].

Objective: To summarize the clinical characteristics and the features of electroencephalograph (EEG) of children with DEPDC5 gene variants related epilepsy. Methods: The clinical data, gene variation, EEG and head magnetic resonance image (MRI) of 20 epileptic children with DEPDC5 gene variants admitted to Department of Pediatrics, Peking University First Hospital from May 2017 to November 2020 were retrospectively analyzed. Results: Twenty patients with heterozygous DEPDC5 gene variants were enrolled, 8 of 20 patients were nonsense variants, 6 were missense variants, 3 were frame-shift variants, 2 were splicing variants, and 1 was large fragment deletion. Sixteen cases had hereditary variation and 4 had de novo variation. Fifteen of variations were novel. Nine were male, while 11 were female. Their latest follow-up age ranged from 10 months to 13 years and one month.The epilepsy onset age ranged from 3 hours to 11 years and 3 months, the median age was 10.5 months. Twelve (60%) patients had developmental delay. Nineteen patients had focal seizures, 7 had epileptic spasms, 1 had multiple seizure types including tonic, atypical absence, dystonic and myoclonic seizures. Epileptic form discharges were observed in 18 patients during the interictal phase, and 11 were focal discharges, 7 were multifocal discharges. Ten (50%) patients had abnormal brain MRI, including focal cortical dysplasia in 5 patients, undefined malformation of cortical development in 4 patients, hemimegalencephaly in 1 patient. Four patients were diagnosed as West syndrome and one patient was diagnosed as Lennox-Gastaut syndrome. Fourteen (70%) patients were diagnosed as drug-resistant epilepsy. Four patients became seizure-free by treatment with anti-epileptic drugs. Three children were treated with surgery, and 2 of them became seizure-free, 1 had more than 75% reduction in seizures. Conclusions: DEPDC5 gene variant epilepsy is inherited with incomplete penetrance and focal seizure is the major seizure type. However, epileptic spasms, generalized seizures can also be observed. Half of the patients brain malformations. Most of the patients are drug-resistant epilepsy. Patients with clear epileptogenic zones can be treated with surgery. Treatment-resistant patients are more likely to be complicated with developmental delay.

[Clinical features of epilepsy in children with IRF2BPL gene variation].

Objective: To summarize the genotype and phenotype of epilepsy in patients with interferon regulatory factor 2 binding protein-like (IRF2BPL) gene variants. Methods: Data of 6 epilepsy patients with IRF2BPL gene variants seen from May 2017 to September 2020 in the Department of Pediatrics of Peking University First Hospital were retrospectively collected. The clinical characteristics and genetic test results were analyzed. Results: A total of 6 patients with IRF2BPL gene variants (1 boy and 5 girls) were identified. The age of seizure onset was from 3.5 to 7.0 months. Epileptic spasms were observed in 6 patients, tonic seizures and tonic-spasms were observed in 1 patient and focal seizure was observed in 1 patient. All 6 patients presented with developmental delay, 5 patients presented with hypotonia, and 2 patients presented with dysphagia. Microcephaly,nystagmus,chorea and athetosis were observed in 1 patient. The electroencephalography (EEG) showed slow background activity in 2 patients. Hypsarrhythmia was observed in all 6 patients. Focal epileptic discharges were observed in 2 patients. Epileptic spasms were monitored in all 6 patients. Focal seizure and tonic-spasm were monitored in 2 patients respectively. The brain magnetic resonance imaging (MRI) showed cerebral atrophy and dysplasia of the corpus callosum in 1 patient, delayed myelination in 2 patients and normal in 3 patients. Two patients had missense variants c.1280C>T/p.L474F and c.1420C>T/p.S427L, 3 patients had frameshift variants c.232delG/p.V78Sfs*73, c.244del/p.A82Pfs*70 and c.283-308del/p.Ala95Thrfs*29, 1 patient had non-frameshift deletion variant c.1453-c.1455delTTC/p.F485del, and all of the 6 cases had de novo variants. All patients were diagnosed with infantile spasms. The last follow-up age ranged from 1 year to 3.8 years. Four patients achieved seizure-free and 2 patients still had frequent seizures after the treatment with antiepileptic drugs (adrenocorticotropic hormone, topiramate, and vigabatrin). Conclusions: IRF2BPL gene variants are mainly de novo. The age of seizure onset is mainly in infancy, and epilepsy and developmental delay are the main clinical manifestations. Infantile spasm is the main phenotype, some patients have hypotonia and dysphagia. Cerebral atrophy can be observed in a few patients.

[Clinical characteristics of SLC35A2 gene variants related congenital disorders of glycosylation typeâ ¡].

Objective: To summarize the clinical characteristics of children with SLC35A2 gene variants related congenital disorders of glycosylation (SLC35A2-CDG), so as to improve the clinicians' understanding of this disease. Methods: Clinical data and gene detection results of 6 epilepsy children with SLC35A2 gene variants were treated in the Department of Pediatrics Peking University First Hospital from April 2019 to February 2020 were analyzed retrospectively. Results: Six children with SLC35A2 gene variants were identified, including 1 male and 5 females. The onset age of seizure was 5.5 (ranged from 2 to 20) months. All 6 cases had epileptic spasms, 2 cases had focal seizures, 2 cases had myoclonic seizures, 1 case had tonic seizures and 1 case had generalized tonic-clonic seizures. All patients with SLC35A2 gene variants were diagnosed as infantile spasm with developmental delay. Four cases had microcephaly, 4 cases had micro skeletal abnormalities, 3 cases had hypotonia and facial dysmorphism, 2 cases had inverted nipples. Visual abnormality, auditory anomaly, congenital cardiac disease and feeding difficulty were observed in one patient. The electroencephalography showed hypsarrhythmia in 6 patients. The brain magnetic resonance imaging (MRI) showed thinning of corpus callosum in 3 patients, delayed myelination in 2 patients and normal brain MRI in 3 patients. There were 2 cases of in-frame deletions, 1 case of missense variant, 1 case of splice site variant, 1 case of 2.14 kb deletion in Xp11.23 (only containing SLC35A2 gene) and 1 case of SLC35A2 gene mosaicism. All 6 cases had de novo variants. The last follow-up age ranged from 18 to 52 months. One patient was seizure free and 5 patients still had frequent seizures after treatment with antiepileptic drugs. Conclusions: SLC35A2 gene variants are mainly de novo variants. The characteristics of patients with SLC35A2-CDG are seizures and developmental delay, infantile spasms are most common diagnosis, micro skeletal anomaly, microcephaly, hypotonia, facial dysmorphism were accompanied features.

MiRNA-221-5p promotes breast cancer progression by regulating E-cadherin expression.

OBJECTIVE: To elucidate the role of miRNA-221-5p in the development of breast cancer (BCa) and its underlying mechanism. PATIENTS AND METHODS: The expression level of miRNA-221-5p in 52 pairs of BCa tissues and adjacent normal tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between miRNA-221-5p expression and pathological indicators of BCa was analyzed. MiRNA-221-5p expression in BCa cells was also determined by qRT-PCR. After transfection of miRNA-221-5p inhibitor in MCF-7 and SKBR3 cells, we detected the regulatory effects of miRNA-221-5p on cellular behaviors through cell counting kit-8 (CCK-8), wound healing, and transwell assay. Finally, the relationship between miRNA-221-5p and E-cadherin in BCa was elucidated. RESULTS: QRT-PCR results showed that the expression level of miRNA-221-5p in BCa tissues was markedly higher than that in normal tissues. Compared with BCa patients with low expression of miRNA-221-5p, those with high expression had a higher incidence of lymph node metastasis and distant metastasis. However, miRNA-221-5p expression did not correlate with age and sex of BCa patients. MiRNA-221-5p was also highly expressed in BCa cells. Transfection of miRNA-221-5p inhibitor suppressed proliferative, invasive, and migratory potentials of BCa cells. Subsequently, we verified that E-cadherin was lowly expressed in BCa cells, and negatively correlated with miRNA-221-5p. In addition, rescue experiments confirmed that transfection of si-E-cadherin reversed the inhibitory role of miRNA-221-5p knockdown in migratory and invasive potentials of BCa cells. CONCLUSIONS: The expression of miRNA-221-5p remained high in BCa, which was correlated with lymph node metastasis, distant metastasis, and poor prognosis of BCa. MiRNA-221-5p may promote the invasive and migratory potentials of BCa by regulating E-cadherin expression.

[Anatomical and finite element analysis of anterior cruciate ligament reconstruction within biomechanical insertion].

OBJECTIVE: To provide new concepts of anterior cruciate ligament (ACL) reconstruction by anatomical gross observation of ACL tibial insertion and finite element analysis of distribution of ACL mechanical insertion. METHODS: In the anatomical study, ten fresh adult cadaveric knees were dissected, including 6 males and 4 females, all knees were generally observed through standard medial parapatellar approaches, paying attention to the close anatomical relationship of tibial insertion and anterior horn of lateral meniscus, and ACL was exposed and gradually removed from the inside. The shape of tibial insertion of ACL was observed and recorded, and anterior-posterior diameters and left-right diameters of tibial insertion were measured with vernier caliper. For the study of finite element analysis, three-dimensional thin-layer magnetic resonance imaging of normal knee joint was used to establish knee joint model. Three-dimensional reconstruction software MIMICS and finite element analysis software ANSYS were used to establish knee joint model, subsequently, clinical physical examination Lachman test and pivot-shift test were simulated to observe the force distribution of ACL tibial insertion and femoral insertion. RESULTS: The ACL tibial mechanical insertion was rather flat and long similar as an arc shape without a clear separation between anterior medial bundle (AMB) and posterolateral bundle (PLB) in gross observation. The dense fibers lies belonged to the medial intercondylar ridge and ended up anterior with the osseous landmark of anterior ridge. Its average anterior-posterior diameter was (13.8±2.0) mm, the average left-right diameter of midsubstance was (5.3±0.6) mm, and the average left-right diameter of anterior margin was (11.5±1.2) mm. The finite element analysis showed that distribution on the femoral side was oval shape mainly below the residents' ridge, while the tibial side was rather flat mainly along the medial intercondylar ridge, which was consistent with the anatomical observation. The biomechanical characteristics of ACL attachments were verified theoretically. CONCLUSION: Anatomical study and finite element analysis have confirmed the flat arc shape of ACL tibial insertion. The ideal reconstruction technique of ACL should be based on its biomechanical insertion. Based on anatomical study and biomechanical analysis, we have proposed the idea of ACL biomechanical insertion reconstruction (BIR) and established a surgical model with oval femoral tunnel and rounded-rectangle tibial tunnel.

Hyperhomocysteinemia Is Associated with Vitamin B-12 Deficiency: A Cross-sectional Study in a Rural, Elderly Population of Shanxi China.

OBJECTIVES: To report the association of lifestyle factors and plasma vitamin B-12 with hyperhomocysteinemia in a large sample of men and women living in a region of China where there is an increased risk of NTDs. DESIGN: Community-based, cross-sectional study of Lvliang City, Shanxi Province, China. SETTING: Hyperhomocysteinemia is an independent risk factor for cardiovascular disease (CVD) and a sensitive marker of vitamin B-12 and folate deficiency. PARTICIPANTS: A total of 2355 (1044 men and 1311 women) participants born before 1 January 1958 (≥55 years of age) and living in Lvliang City for at least 2 months a year were included. MEASUREMENTS: The participants were assessed regarding demographic characteristics, height, weight, as well as having a physical examination and blood sampling for serum cholesterol, total homocysteine (tHcy), folate, and vitamin B12 levels. RESULTS: The median (25th-75th percentile) tHcy concentration was 21.5 (15.8-33.6) µmol/L in men and 18.0 (13.4-24.8) µmol/L in women. The overall prevalence of hyperhomocysteinemia (tHcy ≥15 µmol/L) was 72.6% (84.3% in men and 63.2% in women), inversely correlated with folate (r=-0.230, P=0.006) and vitamin B-12 (r=-0.540, P<0.001), and positively correlated with uric acid (r=0.054, P<0.001). Vitamin B-12 and folate deficiency, older age, and male gender were associated with elevated tHcy; with vitamin B-12 deficiency being the strongest. CONCLUSIONS: Plasma tHcy concentration and hyperhomocysteinemia were significantly higher in this population than in previously studied populations. Vitamin B-12 and folate supplementation, concomitant lifestyle changes such as smoking cessation, and lipid-lowering treatments may help to decrease plasma tHcy concentrations and reduce the CVD risk in this population.

Accumulation of flavanols and expression of leucoanthocyanidin reductase induced by postharvest UV-C irradiation in grape berry.

To examine the effect of postharvest ultraviolet C (UV-C) irradiation on flavanol polyphenol accumulation in the grape berry, we investigated total flavanol polyphenol content, the enzyme activity of leucoanthocyanidin reductase (LAR), and transcription of Vv lar1 and Vv lar2 using spectrophotometry, real-time polymerase chain reaction, and western blot analysis in 5-year-old Vitis vinifera L. cv. Cabernet Sauvignon plants. Our results indicated that the accumulation of flavanol polyphenol reached its highest value when exposed to UV-C irradiation for 30 min. Additionally, UV-C irradiation induced the transcription of Vv lar1 and Vv lar2 and the synthesis of LAR1 and LAR2 proteins, resulting in increased accumulation of flavanol polyphenol in the grape berry. Moreover, these effects were associated with the length of time of UV-C irradiation.

Nutritional status of the elderly in rural North China: a cross-sectional study.

OBJECTIVES: This study aimed to describe the nutritional status of elderly people living in a rural area of North China. DESIGN: Community-based, cross-sectional prevalence survey. SETTING: 3 rural towns of Lvliang City, Shanxi Province, China. PARTICIPANTS: A sample of 1845 community residents (29.1% of those eligible) 55 years or older (birth before 1958-01-01). MEASUREMENTS: The participants were assessed regarding demographic characteristics, height, weight, as well as having a physical examination and blood sampling for serum cholesterol, total homocysteine (tHcy), folate, and vitamin B12 levels. RESULTS: 991 (53.7%) were female and 139 (7.5%) did not complete the anthropometric measurement. Prevalence of underweight and obesity was 3.5% and 24.9% in men and 6.7% and 31.0% in women (P = 0.003, P = 0.005, respectively). Prevalence of hypercholesterolemia and hypocholesterolemia was 13.5% and 52.6% in men and 25.0% and 34.3% in women (P < 0.001, P < 0.001, respectively). Prevalence of high LDL-c concentrations was 8.8% in men and 16.8% in women (P < 0.001). The mean serum tHcy in men (28.8 ± 20.1 µmmol/l) was significantly higher than in women (21.0 ± 15.1 µmmol/L, P < 0.001). Prevalence of hyperhomocysteinemia (defined as > 15µmmol/L) was 79.7% in men and 65.5% in women (P < 0.001). Prevalence of low folate (defined as < 11 nmol/L) and vitamin B12 levels (defiend as < 185 pmol/L) was 70.8 % and 76.8% in men and 56.5% and 72.6% in women (P < 0.001, P = 0.036, respectively). Correlation coefficients between tHcy, folate, and vitamin B12 indicated an inverse linear correlation (r = -0.21, P < 0.001, r = -0.35, P < 0.001, respectively). CONCLUSIONS: As China's economic climate has developed, the nutritional status of elderly people in the rural parts of the country has improved in some aspects. However, the trend toward obesity will lead to a shift in the burden of obesity-related chronic diseases. In addition, rurally-located elderly people are at high risk of death that may be associated with abnormal serum cholesterol. The data also suggest that severe deficiencies in folate and vitamin B12 levels exist, as well as there being a high prevalence of hyperhomocysteinemia. Folate and vitamin B12 supplementation are necessary to prevent related diseases.

Vintage year determination of bottled Chinese rice wine by VIS-NIR spectroscopy.

The feasibility of noninvasive visible and near infrared (VIS-NIR) spectroscopy for determining vintage year of Chinese rice wine was investigated. VIS-NIR spectra of 100 samples were collected in transmission mode in 600-mL square brown glass bottles by a fiber spectrometer system. Discriminant models were developed based on discriminant analysis (DA) together with raw, 1st, and 2nd derivative spectra. The concentration of alcohol content, total acid, and degrees Brix was determined to validate the VIS-NIR results. The calibration result for raw spectra was better than that for 1st and 2nd derivative spectra. The percentage of samples correctly classified for raw spectra was 97.1%. For the sample groups in the vintage years of 2002, 2003, and 2004, the samples were all correctly classified. And for the 2000 and 2001 sample groups, the percentage of samples correctly classified was 92.9%. In validation analysis, the samples were all correctly classified. The results demonstrated that the VIS-NIR spectroscopic technique could be used as a noninvasive and rapid method for predicting vintage year of bottled Chinese rice wine.

Inhibition of HPV 16 E6 oncogene expression by RNA interference in vitro and in vivo.

Substantial studies have demonstrated that the initiation and progression of cervical cancer were closely associated with human papillomavirus (HPV) E6 and E7 oncogenes. The therapeutic strategy with ribozyme or antisense oligonucleotides to inhibit the expression of HPV E6 or E7 oncogenes showed effect to some degree, but problems such as low efficiency, short-period maintenance, and high cost still remain. The aim of this study was to investigate in vitro and in vivo the effect of HPV 16 E6 small interfering RNA (HPV 16 E6 siRNA) on cervical cancer cell line CaSki cells. The specific siRNA of HPV 16 E6 was synthesized and transfected into CaSki cells by liposome. The number of apoptotic cells, HPV 16 E6 messenger RNA (mRNA) level, and E6 protein expression were measured before and after the transfection by flow cytometry, reverse transcriptase-polymerase chain reaction, and Western blot, respectively. Cervical cancer in nude mice was established, and siRNA was injected directly into the nude mice peritoneal cavity or subcutaneous tumor. The efficiency of siRNA was evaluated by tumor volume change, HPV 16 E6 protein expression, and apoptosis of tumor cells. Apoptosis rate of CaSki cells at days 1, 2, 5, and 9 after siRNA transfection were 7.7%, 11.8%, 37.4%, and 12.6%, respectively. The mRNA level of HPV 16 E6 at the same time points were reduced by 77%, 83%, 59%, and 41%, respectively. But the mRNA level of beta-actin, as an internal control, showed no significant change. The inhibition rates of E6 protein synthesis at days 1, 2, 5, and 9 after the transfection were 79.7%, 80.4%, 71.3%, and 57.4%, respectively, whereas the protein levels of Lamin A/C, as internal control, had no change. In vivo, E6 siRNA administration groups showed a dramatic effect in inhibiting tumor growth, suppressing expression of E6 protein, and inducing tumor necrosis and apoptosis as compared with the control group. Direct injection of siRNA into subcutaneous tumor resulted in tumor suppression effect similar to that via the peritoneal cavity, and with additional injection better results could be achieved in cervical cancer CaSki cells. RNA interference exists, and the interference to HPV 16 E6 is specific and highly efficient both in vitro and in vivo.

[Increase of 3H-GABA release from rat cultured cortex neurons after chronic exposure to MK801].

Excitatory amino acids are involved in acute and chronic neurodegenerative diseases. Little is known about the potential consequences of chronic blockade of NMDA receptors (one subtype of excitatory amino acid receptors). Receptor function measured as 3H-GABA release in culture media after pretreatment with MK801 was studied in rat cortical neurons in primary cultures. Cultured neurons were exposed to 1 mumol.L-1 MK801 for 4 days since the 14th day. Glutamate (1 mmol.L-1) evoked 3H-GABA release was shown to be significantly increased (control 0.2174/1000 +/- 1.40/1000; MK801 treatment 0.763% +/- 0.192%). KCl 40 mmol.L-1 stimulation showed no such effect. This result suggests that the NMDA receptor function of releasing neurotransmitters changed after chronic treatment with noncompetitive antagonists.

[Comparison of the affinities of amoxapine and loxapine for various receptors in rat brain and the receptor down-regulation after chronic administration].

Using radioligand receptor binding methods, the affinities (Ki) of amoxapine and loxapine for various receptors (adrenergic alpha 1, alpha 2, beta; dopaminergic D1, D2; serotoninergic 5-HT1, 5-HT2; Muscarinic, GABA, BZ) were investigated. The two compounds showed high affinities for 5-HT2, D2 and alpha 1 receptors (Ki less than 10(-7) mol/L), moderate affinity for alpha 2 receptor (Ki less than 10(-6) mol/L), and low affinities for M and 5-HT1 receptor (Ki less than 10(-5) mol/L). In addition, amoxapine appeared to have low affinities for D1 and GABA receptors. For D1 receptor, loxapine was found to have moderate affinity which was nearly 20 fold greater than amoxapine, but amoxapine exhibited more potent inhibitory effects on serotonin receptors and weaker inhibitory affects on dopamine receptors. Neither amoxapine nor loxapine showed significant affinity for BZ and beta-adrenergic receptors. These differences in the affinities may be responsible for their different psychopharmacological effects in the clinical treatment of patients. The regulation of 5-HT2 and beta receptors were examined in chronic experiments on rats given amoxapine 8 mg/kg or loxapine 1 mg/kg orally once daily for one to three weeks. The 5-HT2 receptor density was time-dependently reduced but no effect on beta receptors was observed. The down-regulation of 5-HT2 receptors might be associated with antidepressant action of the two drugs.

Metabolic fate of Qinghaosu in rats; a new TLC densitometric method for its determination in biological material.

Since the sixties, the emergence of malarial parasites resistant to the most potent anti-malarials has posed a serious problem to the therapy of malaria. Qinghaosu, a new sesquiterpene isolated from a Chinese medicinal herb Qing-hao (Artemisia annua Linn) is being used for the treatment of malaria in China with good results even in cases resistant to common anti-malarial agents. In this paper, a sensitive method of high specificity using TLC for the determination of Qinghaosu in biological specimens and in the study of the metabolism of the drug in rats is described. Qinghaosu was shown to be completely and rapidly absorbed after oral administration. However, a very low plasma level was obtained even after a dose of 300 mg/kg. Liver was found to be the chief site of its inactivation. When Qinghaisu was given intramuscularly, significant and more persistent plasma levels were detected. Qinghaosu was shown to pass the blood-brain and blood-placenta barriers after i.v. injection. Very little unchanged Qinghaosu was found in the urine and feces in 48 hours regardless of administration route (i.v., i.m. or p.o.).