PURPOSE: Diallyl disulfide (DADS), an organosulfur compound in garlic, has been reported to be effective in inhibiting the growth of several human tumor cell lines. The aim of this study was to determine whether DADS induced growth inhibition in MCF-7 breast cancer cell lines and to understand the molecular mechanism by which DADS acts. METHODS: MCF-7 cell lines were incubated with various concentrations of DADS for various time intervals and the cytotoxicity was determined by MTT assay. We examined the changes of intracellular proteins related to apoptosis, such as bcl-2, bax and PARP in cells treated with DADS. To study the expression level of bcl-2 and bax, which serve as modulators of apoptosis, we performed RT-PCR and western blot analysis. RESULTS: MCF-7 cells treated with DADS led to the suppression of viability and proliferation in both a time and concentration dependent manner. Microscopic observation revealed typical features of apoptosis in the DADS-treated cells, further verified in nuclear DAPI staining. Flow cyto-metry analysis with FITC-annexinV and propidium iodide (PI) demonstrated that the apoptotic cell population with AnnexinV+/PI- increased dramatically from ~0.8% to ~75% after 24h exposure to 500 microM DADS in MCF-7 cells. Cell cycle analysis demonstrated that the number of apoptotic cells increased with the increasing time of the DADS treatment. Additionally, thermore, we investigated the effects of DADS on apoptosis related gene expression in MCF-7 cells. PARP cleavage was markedly increased in the DADS treated cells with time. This result indicated that DADS induced the caspase-dependent apoptotic pathway. We also found down-regulation of bcl-2, however no significant change of Bax expression was observed after DADS treatment. Conclusion: Taken together, these results indicate that DADS induces apoptosis by activating a caspase pathway involving the activation of Bcl-2 but not of Bax. Our findings suggest chemotherapeutic potentials of DADS in human breast cancer.
Apoptosis ; Blotting, Western ; Breast Neoplasms* ; Breast* ; Cell Cycle ; Cell Line* ; Cell Line, Tumor ; Down-Regulation ; Garlic* ; Gene Expression ; Humans* ; MCF-7 Cells ; Propidium

Objective To analyze the clinical characteristics,muscle pathological features and pathogenic gene mutation in 4 cases with LMNA-related congenital muscular dystrophy (L-CMD).Methods Clinical data of the probands and the parents were collected.Skeletal muscle specimens were biopsied from the probands for pathological analysis.Genomic DNA and RNA were extracted from peripheral blood leukocytes,and PCR,reverse transcription(RT)-PCR and DNA direct sequencing were employed to analyze the LMNA gene to determine the gene mutation and confirm the pathogenicity.Results Four patients had symptoms from fetal period to several months after birth.They presented with motor retardation,muscle weakness with prominent the proximal upper limbs,distal lower limbs and neck extensor,hypotonia,contractures,with mild to moderate elevation of CK level.The muscle biopsies showed muscular dystrophic and with inflammatory changes,and the abnormal nuclear morphology was observed with transmission electron microscopy.Genetic analysis of them detected 4 dominant de novo mutations.Three of them had unreported pathogenic mutations.The same sites of the LMNA gene were wild type in their parents.Conclusions Four cases of L-CMD are genetically identified.Genetic counseling of the family can be possible.The patients should be considered LMNA gene mutation of they present themselves with muscle weakness with the proximal upper limbs,distal lower limbs and neck extensor,hypotonia,contractures,mild to moderate elevation of CK level,and if the biopsies show muscular dystrophic changes but also with inflammatory changes should be considered LMNA gene mutation.Genetic analysis is the most reliable method for diagnosing L-CMD.

OBJECTIVE: To study the clinical characteristics of epileptic patients with synchronous occipital and frontopolar spikes phenomenon. METHODS: The clinical data,EEG features of patients with synchronous occipital and frontopolar spikes phenomenon were retrospectively reviewed. RESULTS: Totally 34 male and 21 female patients had synchronous occipital and frontopolar spikes. Ages at the time of recording the synchronous occipital and frontopolar spikes phenomenon ranged from 2 years 6 months to 14 years with the median age of 6 years 6 months. In the total 55 patients,12 patients had contralateral synchronous discharges,12 patients had ipsilateral synchronous,17 patients had bilateral synchronous discharges,7 patients had bilateral synchronous discharges tending to contralateral synchronously,and 7 patients had bilateral synchronous discharges tending to ipsilateral synchronously. Occipital preceded frontopolar discharges in 42 patients. Frontopolar preceded occipital discharges in 3 patients. The phase difference between the occipital and frontopolar spikes could not be distinguished in 10 patients. Based on the etiology and diagnoses,there was Panayiotopoulos syndrome in 33 patients,epilepsy with structural abnormality in 13 patients,secondary epilepsy due to immune encephalitis in 1 patient,epilepsy accompanied by inborn error of metabolism in 2 patients and epilepsy with unknown etiology in 6 patients. CONCLUSION: The synchronous occipital and frontopolar spikes phenomenon usually appears during childhood or develops from other discharge patterns in infancy stage. The synchronous occipital and frontopolar spikes phenomenon can be ipsilateral synchronous or contralateral synchronous discharges. The occipital spikes preceded frontopolar spikes in most patients. The synchronous occipital and frontopolar spikes phenomenon is mostly found in Panayiotopoulos syndrome,but it is also found in epilepsy with different etiologies.

OBJECTIVETo study the effect of skin-derived progenitor cell (SKP) combined with hyaluronic acid( HA) on the wound healing in diabetic rats.

METHODSSKP of Spraque-Dawley (SD) neonate rats were isolated and cultured and mixed with HA. The differentiation characteristics of SKP in the culture were observed. Sixty SD rats were injected intraperitoneally with 65 mg/kg streptozotocin( STZ) to induce diabetes. Two symmetrical full-thickness cutaneous wounds( 1.0 cm in diameter) were made on the back of each SD rat and randomly divided into A (n = 20, with treatment of 100 mircol SKP-HA) , B (n = 20, with treatment of 100 mirol HA) , and C ( n = 20, with treatment of DMEM/F12 culture medium) groups. Tissue samples from wound in each group were harvested on 1, 2, 3, 4 weeks after the treatment. Wound healing rate, changes in histomorphology, the content of hydroxyproline ( HYP) , and immigration of labelled SKP were determined and analyzed.

RESULTSSKP grew well when cultured with HA. The characteristics of SKP to differentiate into lipocyte, neuron, and neurogliocyte remained in the culture. Compared with that in C group, epithelization in the wounds of A and B groups appeared earlier. The wound healing rate in A group [ (72.1 +/- 2. 8)% ] and B group [ (53.7 +/- 2. 9)% ] were obviously higher at 2 post-treatment weeks(PTW) than that in group C [(42. 5 +/- 1.5)% ( P <0.05) , and that in A group was obviously higher compared with B and C groups at 3 PTW ( P < 0. 05 or 0. 01). The wound healing rates in A and B groups were (100. 00 +/- 0.00) % at 4 PIW, which were obviously higher than that of group C( P <0.01) . There was no obvious difference in the HYP content among the 3 groups at 1 PIW ( P > 0. 05) , but it was obviously higher in A and B groups than that in C group at 2,3,4 PTW( P <0.01) , and that in A group was significantly higher than that in B group at 3 and 4 PTW( P <0. 01). SKP survived well on the wound, and migrated towards the dermis as time elapses.

CONCLUSIONSKP-HA composition can promote wound healing in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Hyaluronic Acid ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Skin ; cytology ; Stem Cells ; chemistry ; cytology ; Wound Healing ; drug effects

OBJECTIVETo establish an advanced purification techniques of the essential oils obtained from Pogostemon cablin.

METHODMolecular distillation (MD) was applied.

RESULTFour distillates were obtained, chemical constituents of which were analyzed with GC-MS. Compared with those in original oils, the contents of active compounds (patchouli alcohol and pogostone) rose by 27%-47% in the distillates II and III.

CONCLUSIONMolecular distillation (MD) effectively raises the contents of patchouli alcohol and pogostone. The work is of great economic and scientific significance for the industrialization of P. cablin and the discovery of new drugs.

Gas Chromatography-Mass Spectrometry ; Lamiaceae ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Sesquiterpenes ; isolation & purification ; Technology, Pharmaceutical ; methods

OBJECTIVE: To investigate whether the tonic-clonic seizure (TCS) induced by intermittent photic stimulation (IPS)was generalized tonic-clonic seizure (GTCS)or partial secondarily tonic-clonic seizure (PGTCS),and to analyze the relationship between them. METHODS: Video-electroencephalogram (VEEG)database of Peking University First Hospital from March 2010 to October 2018 were reviewed. Fifteen cases with idiopathic epilepsy who had TCS induced by IPS were included in this study, and their clinical and electroencephalogram (EEG)characteristics were retrospectively analyzed. RESULTS: In this study, 4 of the 15 cases were boys and 11 were girls. The age of seizure onset ranged from 1 to 13 years. According to the medical records: 12 cases were considered as GTCS,while the remaining 3 cases were considered as PGTCS. The age at VEEG monitoring ranged from 2.5 to 16.0 years. All backgrounds of the VEEG were normal. Interictal discharges:generalized discharges in 11 cases, of which 4 cases coexisted with posterior discharges, 2 cases coexisted with Rolandic discharges, the other 5 cases merely had generalized discharges; merely focal discharges in two cases, one in the Rolandic area and the other in the posterior area; no interictal discharge in the remaining 2 cases. IPS induced photoparoxysmal response (PPR)results: 2 cases without PPR,the remaining 13 cases with PPR of generalized discharges, and 6 of the 13 cases coexisted with posterior discharges. IPS induced photoconvulsive response (PCR)results: GTCS in one case (contradictory to medical history),PGTCS in 11 cases (consistent with medical history),and GTCS and PGTCS hardly to distinguish in the remaining 3 cases. Of the three conditions above, there were generalized myoclonic seizures induced by IPS before TCS in 7 cases. CONCLUSION The medical history was unreliable in determining whether TCS was generalized or focal. Myoclonic seizures can coexist with PGTCS, and sometimes GTCS was indistinguishable from PGTCS, indicating that the dichotomy of seizure types need to be improved. Photosensitive TCS should be regarded as a continuum between focal and generalized seizures.
Electroencephalography ; Epilepsy, Generalized ; Epilepsy, Tonic-Clonic ; Female ; Humans ; Male ; Retrospective Studies ; Seizures

PURPOSE: Diallyl disulfide (DADS), an organosulfur compound in garlic, has been reported to be effective in inhibiting the growth of several human tumor cell lines. The aim of this study was to determine whether DADS induced growth inhibition in MCF-7 breast cancer cell lines and to understand the molecular mechanism by which DADS acts. METHODS: MCF-7 cell lines were incubated with various concentrations of DADS for various time intervals and the cytotoxicity was determined by MTT assay. We examined the changes of intracellular proteins related to apoptosis, such as bcl-2, bax and PARP in cells treated with DADS. To study the expression level of bcl-2 and bax, which serve as modulators of apoptosis, we performed RT-PCR and western blot analysis. RESULTS: MCF-7 cells treated with DADS led to the suppression of viability and proliferation in both a time and concentration dependent manner. Microscopic observation revealed typical features of apoptosis in the DADS-treated cells, further verified in nuclear DAPI staining. Flow cytometry analysis with FITC-annexinV and propidium iodide (PI) demonstrated that the apoptotic cell population with AnnexinV /PI increased dramatically from ~0.8% to ~75% after 24h exposure to 500nM DADS in MCF-7 cells. Cellcycle analysis demonstrated that the number of apoptotic cells increased with the increasing time of the DADS treatment. Additionally, thermore, we investigated the effects of DADS on apoptosis related gene expression in MCF-7 cells. PARP cleavage was markedly increased in the DADS treated cells with time. This result indicated that DADS induced the caspase-dependent apoptotic pathway. We also found down-regulation of bcl-2, however no significant change of Bax expression was observed after DADS treatment. CONCLUSION: Taken together, these results indicate that DADS induces apoptosis by activating a caspase pathway involving the activation of Bcl-2 but not of Bax. Our findings suggest chemotherapeutic potentials of DADS in human breast cancer.
Apoptosis* ; Blotting, Western ; Breast Neoplasms* ; Breast* ; Cell Line* ; Cell Line, Tumor ; Down-Regulation ; Flow Cytometry ; Garlic* ; Gene Expression ; Humans* ; MCF-7 Cells ; Propidium

Objective To analyze the clinical characteristics,muscle pathological features and pathogenic gene mutation of a family with autosomal dominant Emery-Dreifuss muscular dystrophy (AD-EDMD).Methods Clinical data of the proband and her family members,a Chinese family of AD-EDMD,were collected.Skeletal muscle specimens were collected from the proband for pathological analysis.Genomic DNA from the proband and her parents was extracted using standard procedures from the peripheral blood leukocytes.PCR and DNA direct sequencing were employed to analyze all of the 12 exons of the LMNA gene to determine the gene mutation,and the case was summarized along with related literature review.Results The proband,female,4 years and 5 months old now,presented with muscle weakness during her early childhood,the proximally was more prominent,mild pectus excavatum.Her CK level was elevated,her electromyogram showed myogenic injuries,the muscle biopsy showed myopathy changes.Her father had the same symptom,with disease progressed,showed elbow contractures in early stage,stiff neck,tight achilles tendon,slowly progressive muscle weakness of the limbs,sinus bradycardia.A heterozygous missense mutation c.1580G ＞ C (p.Arg527Pro) was identified in exon 9 of the LMNA gene in the proband and her father,but not in her mother.This heterozygous missense mutation had been reported as a pathogenic gene mutation.Conclusions The patient who has elbow contractures in early stage,limited neck flexion,spine stiffness,muscle weakness with the proximal upper limbs and distal lower limbs,and arrhythmia,should have an analysis of the LMNA gene.It's important for the early diagnosis of EDMD,assessment of the prognosis,timely and effectively monitoring the changes of arrhythmia,then taking interventions to improve the quality of life and prolong life.So genetic analysis is most reliable method to diagnose EDMD.

OBJECTIVE: To investigate the clinical significance of high-frequency oscillations (HFOs) on scalp electroencephalography (EEG) in patients with epileptic encephalopathy with continuous spike-and-wave during sleep (CSWS). METHODS: Twenty-one CSWS patients treated for epilepsy from January 2006 to December 2016 in Pediatric Department of Peking University First Hospital were enrolled into the study. Selected clinical variables including gender, age parameters, seizure frequencies and antiepileptic drugs were compared between (a). HFO-positive group and HFO-negative group before methylprednisolone treatment and (b). excellent seizure outcome group and not-excellent seizure outcome group after methylprednisolone treatment. Interictal HFOs and spikes in pre- and post-methylprednisolone scalp EEG were measured and analyzed. RESULTS: Before methylprednisolone treatment, there were 12 of 21 (57%) CSWS patients had HFOs, with a mean value 43.17 per 60 s per patient. The 12 patients with HFOs tended to have more frequent epileptic negative myoclonus/atonic/myoclonus/atypical absences than those without HFOs in a month before methylprednisolone treatment. A total of 518 HFOs and 22 592 spikes were found in the pre-methylprednisolone EEG data of 21 patients, and 441 HFOs (86%) were associated with spikes. The highest amplitudes of HFOs were significantly positively correlated with that of spikes (r=0.279, P<0.001). Rates reduced by methylprednisolone treatment were statistically significant for both HFOs (P=0.002) and spikes (P=0.006). The percentage of reduction was 91% (473/518) and 39% (8 905/22 592) for spikes and HFOs, respectively. The percentage of spike and HFOs changes was respectively 100% decrease and 47% decrease in the excellent seizure outcome group, and they were 79% decrease and 18% increase in the not-excellent seizure outcome group. CONCLUSION Prevalence of HFOs might reflect some aspect of epileptic activity. HFOs were more sensitive to methylprednisolone treatment than spikes and had a good correlation with the prognosis of seizures, and HFOs could be applied to assess epilepsy severity and antiepileptic therapy.
Anticonvulsants/therapeutic use* ; Child ; Electroencephalography/methods* ; Epilepsy/physiopathology* ; Epilepsy, Absence ; Humans ; Methylprednisolone ; Scalp ; Seizures ; Sleep