Child, Preschool ; Dwarfism ; congenital ; diagnosis ; Humans ; Male ; Osteochondrodysplasias ; congenital

The protoplasts of Micromonospora purpurea,the high yield strains of gentammicin producer were mutagenized by diethyl sulfate(DES)and ultraviolet radiation(UV)respectively,then fused,screened by gentamicin resistance and regenerated.The average fermentation unit 2200?U/ml could be achieved by shake flask for 10 batches.The average fermentation unit 1900?U/ml could be obtained by 5L fermentor for 7 batches.The quality of the end product conformed to CP2000,BP2000 and USP26 pharmacopoeia.

BACKGROUND:Pathogenesis of Parkinson’s disease is not completely understood, and there is yet no effective therapy that can prevent the neurodegenerative process of the disease fundamentaly. OBJECTIVE:To explore the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on lactacystin-induced Parkinson’s disease dopaminergic PC12 cel apoptosis and its molecular mechanism. METHODS: Under induction by nerve growth factors, PC12 cels differentiated into dopaminergic neurons, and then were treated with different concentrations of lactacystin for different time. When the cel survival rate was about 50%,the concentration and action time oflactacystin were selected to establish cel models of Parkinson’s disease. In the study, there were control group, lactacystin group, PACAP1-27 group (intervention group 1) and PACAP1-27+PACAP6-27 co-intervention group (intervention group 2). Changes of cel morphology were observed under inverted microscope; cel viability was detected with MTT method; the expression of endoplasmic reticulum stress specific protein caspase-12 was detected by western blot. Then the action of PACAP1-27 and PACAP6-27 to the cytoxicity of lactacystin was observed. RESULTS AND CONCLUSION: With different concentrations and action time of lactacystin, the viability of PC12 cels presented a concentration- and time-dependent decline. When the lactacystin at 20μmol/L acted for 24 hours, the cel viability was declined by about 50%. Under same conditions of lactacystin concentration and action time (20 μmol/L, 24 hours), the cels in the lactacystin group appeared to have damaged changes, declined cel viability, and increased caspase-12 activity in comparison with the control group (P< 0.01). Compared with the lactacystin group, the cel damage was relieved and cel viability was increased significantly in the intervention group 1 as wel as the expression of caspase-12 was decreased (P < 0.01). Experimental findings in the intervention group 2 were similar to those in the lactacystin group. These results suggest that lactacystin, an ubiquitin proteasome inhibitor, can lead to cel damage; PACAP1-27 plays a protective role by regulating the above-mentioned signal pathway. As one PACAP1-27 receptor antagonist, PACAP6-27 can attenuate this effect of PACAP1-27.

Objective:To investigate the feasibility of real-time screening and diagnosing fundus diseases with Wild Field Imaging System (WFIS SW-8000) in the eyes with dense cataract patients.Methods:A series of case-observational study was carried out.Ninety-six dense cataractous eyes of 90 patients with suspected fundus diseases were included in Affiliated Hospital of Nantong University from April to July 2019.Lens phacoemulsification combined with intraocular lens implantation was performed after the opacity lens was removed, and fundus examination was performed with WFIS SW-8000.Super field lens, optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) was used to verify the fundus findings after surgery, which were compared with WFIS SW-8000.This study protocol was approved by Ethics Committee of Affiliated Hospital of Nantong University and followed the Declaration of Helsinki.Written informed consent was obtained from each patient prior to any medical examination.Results:Among 96 eyes, fundus diseases were detected in 40 eyes with the detection rate of 41.67% by WFIS SW-8000 examination, including dry age-related macular degeneration (AMD) in 2 eyes, wet AMD in 3 eyes, polypoidal choroidal vasculopathy (PCV) in 1 eye, high myopia retinopathy (HMR) in 7 eyes, central retinal vein occlusion (CRVO) with macular edema (ME) in 1 eye, mild non-proliferative diabetic retinopathy (NPDR) in 12 eyes, moderate NPDR in 7 eyes, severe NPDR in 4 eyes and proliferative diabetic retinopathy (PDR) in 3 eyes.Fundus diseases were detected in 45 eyes with detection rate of 46.88% after surgery by a super field lens, OCT and FFA.The detectable rate of digital retinal camera examination and super field lens, OCT and FFA showed no significant difference between intraoperation and postoperation (χ 2=0.528, P=0.468). Twenty-two eyes with fundus neovascular disease or macular edema requiring drug intervention were identified intraoperatively, and intravitreal anti-inflammatory and/or anti-vascular endothelial growth factor (VEGF) drugs were injected in 11 eyes during the operation. Conclusions:WFIS SW-8000 is a useful tool for the accurate and convenient method in real-time fundus examination during phacoemulsification, which is feasible and helpful for timely intervention in and treatment of fundus diseases.

OBJECTIVETo discuss the epidemiological and clinical characteristics of the hospitalized children with hand foot and mouth disease (HFMD) in Yantai area.

METHODSEpidemiological and clinical data of HFMD children from 2009 to 2010 were summarized and analyzed retrospectively.

RESULTSMost of the infected (94.6%) were under 5 years old and the ratio between male and female was 1.5: 1. Oral mucosal pox or ulcer as well as hand and foot rashes were observed in all 931 patients. Fever and neurological disorders occurred in 840 (90.2%) and 121 (13.0%) patients respectively. The incidence was positively correlated with air temperature (r = 0.887, P < 0.001), with a peak in April to September (88.9%). The ratio of children from countryside, total duration of fever, serum concentration of c-reacting protein (CRP) and fasting blood glucose (FBG) were significantly higher in severe cases than in those mild ones. Multivariate analysis showed longer mean duration of fever( Odds ratio [OR], 1.491; 95% confidence interval [ CI] 1.170-1.901; P = 0.001) and hyperglycemia (OR, 1.124; 95% CI 1.016-1.245; P = 0.024) were independent risk factors of severity.

CONCLUSIONChildren younger than 5 years old are susceptible to HFMD and most cases occur in April to September. The monthly incidence is positively correlated with temperature of that month. Longer duration of fever and hyperglycemia are independent risk factors for severity. Most cases could have a favorable prognosis after timely diagnosis and proper intervention.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Epidemiologic Studies ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Incidence ; Infant ; Male ; Retrospective Studies ; Seasons ; Temperature

OBJECTIVETo observe the mitochondrial fragmentation and the expression of mito-fusion 1 gene in the cortical neurons of rats with chronic fluorosis, and to reveal their roles in mitochondria damage to neurons due to chronic fluorosis.

METHODSSD rats were divided randomly into three groups of 20 each (a half females and a half males housed individually in stainless-steel cages), and fed with the different doses of fluoride containing in drinking water (untreated control containing 0 mg/L fluoride, and low-fluoride and high supplemented with 10 and 50 mg/L fluoride, respectively). After 3 or 6 months exposure, the mitochondrial morphology of the neurons in rat brains were observed by transmission electron microscopy (TEM), then the expression of mitochondrial fusion gene, Mfn1, were detected by immunohistochemistry and western-blotting, respectively.

RESULTSDental fluorosis was obvious in the rats exposed to excessive fluoride in their drinking water, that is, (16 rats out of 20) numbers of I° detal fluorosis in the low-fluoride group, and (11 rats out of 20) numbers of I° and (9 rats out of 20) numbers of II° detal fluorosis in the high-fluoride group were observed after 3 months exposure. Moreover, (14 rats out of 20) numbers of I° and (6 rats out of 20) numbers of II° detal fluorosis in the low-fluoride group and (6 rats out of 20) numbers of Io, (13 rats out of 20) numbers of II°, and (1 rats out of 20) numbers of III° detal fluorosis in the high-fluoride group were observed after 6 months exposure. And both of untreated controls without detal fluorosis were also observed. The urinary level of fluoride in the low-fluoride group (3.30 ± 1.18) mg/L and in the high-fluoride group (5.10 ± 0.35) were observed after 3 months exposure (F = 3.18, P < 0.05). Moreover, the urinary level of fluoride in the low-fluoride group (4.16 ± 1.39) mg/L and in the high-fluoride group (5.70 ± 1.70) mg/L were also observed after 6 months exposure (F = 3.17, P < 0.05). The normal mitochondrial morphology of neurons in rats without fluorosis was observed after 3 and 6 months, while the abnormal mitochondrial morphology of neurons with fluorosis was shown, presenting mitochondrial fragmentation with swollen cristae and even the fragmented, shortened or stacked punctuate membranes (section observation of three bullous mitochondrial-mitochondrial fission process) by TEM. As compared with controls (53.0 ± 4.54 and 1.21 ± 0.18) at the experiment period of 3 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 51.09 ± 6.25) and western-blotting (1.22 ± 0.26) were no significant difference for low fluoride group (t = 1.7, 1.1, P > 0.05); Mif1 protein analysis with immunocytochemical (the numbers of positive cells: 59.71 ± 5.64) and western-blotting (1.66 ± 0.20) were significantly increasing for high fluoride group (t = 2.1, 2.1, P < 0.05). As compared with controls (36.43 ± 4.04 and 1.00 ± 0.13) at the experiment period of 6 months, Mif1 protein analysis with immunocytochemical (the numbers of positive cells 20.05 ± 4.55 and 17.10 ± 3.86) and western-blotting (0.64 ± 0.08 and 0.39 ± 0.06) were significantly decreasing for the two fluoride group (t = 2.1, 2.2; 2.2, 2.2 respectively, all P value were < 0.05).

CONCLUSIONSTaking excessive amount of fluoride might result in the mitochondrial fragmentation for the changed expression of Mfn1, and the neurons damage from the chronic fluorosis might be associated with the dysfunction of mitochondrial fusion.

Animals ; Drinking Water ; chemistry ; Female ; Fluoride Poisoning ; metabolism ; pathology ; Fluorosis, Dental ; metabolism ; Male ; Membrane Proteins ; metabolism ; Mitochondria ; pathology ; Mitochondrial Proteins ; metabolism ; Neurons ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

AIM To investigate the equilibrium solubilities,oil-water partition coefficients and in vitro skin permeation features of brucine and strychnine in total alkaloids from Strychni Semen.METHODS Saturated dissolution method was applied to determining the equilibrium solubilities of two constituents in ethanol (10％,20％,30％,60％,90％,anhydrous ethanol),trichloromethane,n-octanol and surfactants (0.5％ tween,0.5％ sodium deoxycholate,0.5％ oleic acid).Shake-flask method was adopted in detecting their oil-water partition coefficients in PBS (pH 2.5,4.0,5.0,5.8,6.8,7.0,7.4,9.0).Modified Franz diffusion cell method was used for evaluating their in vitro skin permeation features in PBS,20％ ethanol and anhydrous ethanol.RESULTS Both brucine and strychnine showed the highest equilibrium solubilities in trichloromethane and the lowest equilibrium solubilities in surfactants.The equilibrium solubility of strychnine was higher than that of brucine in ethanol (＞ 20％) or PBS (pH ＜ 8.0),which reached the highest in 60％ ethanol and pH 2.5 PBS,respectively.The similar oil-water partition coefficients of two constituents,proportional to pH value,reached the highest at pH9.0.And they exhibited the highest accumulated transdermal absorptivities in anhydrous ethanol and pH 9.0 PBS,respectively.CONCLUSION Solvent type has obvious effects on the equilibrium solubilities,oil-water partition coefficients and in vitro skin permeation features of both brucine and strychnine.This study can provide a reference for the bioavailability improvement of transdermal drug delivery and development of related preparations.

OBJECTIVETo study the changes of rat red blood cells on T, B lymphocytes proliferation in chronic unpredictable mild stress (CUMS).

METHODSThirty Wistar rats were averagely divided into control group and the CUMS group. The relevant behavioral performance and red blood cells on lymphocytes proliferation Changes by MTT colorimetric were observed.

RESULTSAfter fourteen days, the growth of weight and organic coefficient of brain decreased in model group (P < 0.05), with a significantly reduced consumption and preference of sucrose solutions, and increased pure water consumption as compared with control group (P < 0.05). Plasma corticosterone levels peaked at seven days but on a declining trend after fourteen days. The rate of red blood cells on T, B lymphocyte proliferation in depression group was lower than non-stress in 7, 14, 21 days (P < 0.05).

CONCLUSIONThe repeated chronic mild stress stimulation can induce long-time changes in acts and activities and decrease the rate of red blood cells on T, B lymphocytes proliferation, which can provide a new experimental method to observe the red blood cell immune function under the stress.

Animals ; Cell Proliferation ; Depression ; blood ; etiology ; Disease Models, Animal ; Erythrocytes ; cytology ; Lymphocyte Activation ; Lymphocytes ; cytology ; Male ; Rats ; Rats, Wistar ; Stress, Psychological ; physiopathology

Objective To observe the circadian rhythm of blood pressure and investigate the impact of Norvasc and Tanatril administrated at different time points based on the theory of time therapeutics in hypertensive patients. Methods The 24-hour ambulatory blood pressure was monitored in 130 inpatients and outpatients with grade 3 or 2 hypertension from May 2008 to November 2009. A total of 34 dipper hypertensive patients were grouped into dipper blood pressure group and 5 mg/d of Norvasc and Tanatril were taken by them at 6:00 AM. A total of 96 nondipper hypertensive patients were further divided into three subgroups after adjustment for age and gender: 5 mg/d of Norvasc and Tanatril were taken by group I (n = 30) at 6:00 AM; 5 mg/d of Tanatril at 6:00 AM and 5 mg/d of Norvasc at 18:00 PM by group Ⅱ (n = 32); 5 mg/d of Norvasc at 6:00 AM and 5 mg/d of Tanatrilat 18:00 PM by group Ⅲ(n=34).The 24-hour ambulatory blood pressure monitoring was performed again after four weeks treatment and 24-hour mean systolic blood pressure(24 hSBP),24-hour mean diastolic blood pressure(24 hDBP),daytime and nighttime mean systolic blood pressure(dSBP,nSBP)and daytime and nighttime mean diastolic blood pressure(dDBP,nDBP),were analyzed.Results The 24 hSBP,24 hDBP and dSBP,and dDBP were reduced from(154.3±5.6),(95.4±3.1),(158.7±6.5),(99.6±3.7)mmHg to(137.2±3.9),(82.5±2.7),(139.7±3.8),(85.2±3.5)mmHg,respectively,in dipper blood pressure group(t=2.124,2.356,2.278,2.449,all P<0.05).The 24 hSBP and 24 hDBP of the three groups in nondipper blood pressure were decreased from(154.4±6.1),(156.7±6.7),(156.6±5.2),(95.8±2.8),(94.9±3.8),(95.7±3.2)mmHg to (139.6±4.1),(134.5±4.6),(133.4±3.5),(83.5±4.2),(80.8±5.6),(81.6±4.7)mmHg,respectively(t=2.038,2.040,2.135,2.142,2.213,2.216,all P<0.05).dSBP and dDBP were decreased from(158.6±3.50),(158.4±5.6),(159.5±4.),(98.4±3.7),(99.6±3.7),(83.9±5.2)mmHg to(138.9±5.4),(136.7±4.1),(137.4±6.4),(85.8±5.3),(83.6±5.1),(83.9±5.2)mmHg,respectively(t=2.021,2.252,2.261,2.217,2.167,2.076,all P<0.05).nSBP and nDBP were decreased from(146.7±6.9),(149.8±3.9),(150.2±4.1),(93.7±4.2),(95.7±4.3),(93.4±3.3)mmHg to(133.7±4.6),(129.8±5.7),(127.6±2.8),(87.8±2.9),(78.5±6.4),(77.8±4.8)mmHg,respectively(t=1.798,2.032,2.014,1.864,2.157,2.166,all P<0.05).There were significant differences in nSBP and nDBP among all groups after treatment(F=2.32,2.17,all P<0.05),and the effect of the drugs was better in groug Ⅱ and Ⅲ than in group Ⅰ(q=3.17,4.03,3.32,4.19,all P<0.05),but there were no significant differences between group Ⅱ and Ⅲ(P>0.05).Conclusions Blood pressure can be controlled effectively by taking two antihypertensive medictions in the morning in hypertensive dipper patient but the blood pressure of nondipper hypertensive patients were able to be controlled more efficiently by taking the two antihypertensive medictions separately in the morning and at the evening compared with that taking the two drugs together in the morning.

Background Leber hereditary optic neuropathy (LHON)is a common inherited eye disease,which generally affects young adults with bilateral loss of central vision.Mutation frequency of Leber hereditary has not been fully clarified. Objective This study was to investigate the mutation frequency of mitochondrial NDI gene associated with LHON in Chinese population. Methods The proposal of the study was approved by Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject initial of this trial.Eight hundred and ninety-four LHON patients and 134 normal subjects were collected.Genomic DNA was extracted from peripheral blood leukocytes of the all participants.Polymerase chain reaction (PCR) was used to amplify and sequence analysis of the mitochondrial ND1 gene was performed and aligned with revised Cambridge Reference Sequence(rCRS) of mitochondrial DNA.Then mutated gene frequency was screened and analyzed. Results Mutational analysis of mitochondrial ND1 gene in 894 LHON patients revealed the presence of G3316A,T3394C,G3460A,C3497T,G3635A,G3733A,and T4216C.11.19％ LHON patients (100/894 ) were found to be associated with the gene mutations mentioned above,and 3.24％ patients (29/894) showed the co-occurrence of three primary mutations.Mutation frequencies in LHON patients were 2.57％,2.23％,1.45％,3.80％,0.67％,0.11％,0.34％,respectively,and G3316A,T3394C,C3497T and T4216C also were detected in 134 normal controls with the mutation frequencies of 4.48％,2.99％,4.48％ and 1.49％,respectively.Mutation frequency analysis showed an insignificant difference in the mutations of G3316A,T3394C,C3497T and T4216C between LHON patients and normal controls (x2 =0.926,P=0.336;x2 =0.052,P=0.820; x2 =0.142,P=0.707;P=0.129).G3376A,G3496T,G3700A,A4136G,T4160C and C4171A were absent in Chinese LHON patients. Conclusions Mitoehondrial ND1 gene in LHON is a mutational hotspot in Chinese population,11.19％ (100/894)associated with LHON was caused by ND1 gene mutation.G3635A,G3733A may be rare pathological mutation in Chinese population.However,G3316A,T3394C,C3497T and T4216C are insufficient to produce the clinical phenotype,but they may play a synergic role for penetrance and phenotypic manifestation in LHON.