The effects of cytochalasin D (CD), isolated from Engleromyces goetzii found in our country, on cells of established human esophageal cancer line (ECa-109 cell line) were investigated. It was demonstrated that CD was capable of inhibiting the growth of the cells at the concentration of 0.2?g/ml as indicted by growth curve. The action of CD, however, is reversible, and the cells will gradually grow again normally following the withdrawal of CD. The result also indicated that some proportion of the cells became binucleated or multinucleated after CD treatment. And as the duration of exposure time to CD increased. the multinucleated cells increased in number also. At the concentration of 0.5?g/ml of CD, a phenomenon of extrution of nucleus can be seen among the cells. The nucleus is departed from the cell body but still linked by a fine cytoplasm bridge (Fig. 2). The CD-treated cells bacame enucleated when they subjected to centrifugation following CD treatment. The minicells obtained from enucleation appeared to be normal morphologically with nucleoli but retained only a little cytoplasm around the nucleus. Electromicroscopic observation showed that mitochondria and tonofibrils increased after CD treatment, but we failed to find any changes of microfilaments within the cells.

An“agar-island organ culture”technique is described.It consists of both agar substratum and liquid medium in the same culture dish,so that chemicals in different concentrations can be added into the liquid portion of the medium,a convenient method to test their effects on explants.The procedures are as follows: To one gram of agar in a flask,50 ml of aqua distillata is added,and then heated until the agar is completely dissolved.Then add in 50 ml of double-strength 199 solu- tio(?) into the flask.After shaking gently,mixing the media completely,pour it into a dish until it reaches 2/3 of its height.A few minutes later the agar will solidify.Cut off most of the agar substratum and leave enough agar to form“islands”(Eig.A,B).Then add in liguid medium which is composed of 8 vol 199 solution,2 vol calf serum,2 vol extract of 9-day chick embryo diluted into 50% in Hank's solution and a few drops of diluted penicillin and streptomycin.The explants are placed on the agar-islands,the culture dishes covered with glass tops,sealed with paraffin,are put into an incubator.The cul- ture dishes are not sealed they are put into an incubator supplemented with 5%CO_2 Using this technique,we have cultured 80 explanls of 13-day chick embryo meta- tarsal skin for 7～21 days.And the effect of vit.A with different concentrations are tested The results show that the effects of vit,A on the differentiation of the chick embryo metatarsal skin explants are different with the different concentrations.We believe that this technique can be much more convenient as a method to study the eff- ects of chemicals in different concentrations on explants.The other uses of this tech(?)i- que,such as carcinogenesis in culture,the study of embryonic induction,especially the diffuse mechanism of embryo induction are also briefly discussed.

Objective To investigate the molecular epidemiology of human immunodefieiency viruses (HIV)-1 infected individuals in Honghe district,Yunnan Province and provide the evidence of molecular biology features of HIV-1 infection.Methods HIV-1 pol gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).Then sequencing and phylogenetic tree analysis were employed tO determine HIV-I subgenotype.The sequence alignment was performed in the database of international drug resistance tO identify resistance-associated mutations.Results The samples from 60 HIV-1 infected individuals were investigated:39 were male,21 were female,with average age 35.5 years old.Thirty-four cases were infected with HIV-I through intravenous drug abuse,12 by sexual contacts,2 were contaminated blood/blood products transfusion and 12 with unknown transmission routes.Phylogenetic analysis revealed that 53 cases (88.3%) were subtype 08-BC,6 (10.0%) were subtype 07-BC and 1 (1.7%) was subtype 01_AE.The total rate of drug resistance associated mutations was 33.3%.The major mutations in protease (PR) and reverse transcriptase (RT) regions accounted for 5.0% and 3 1.7%,respectively.The major mutations in PR region were I541M,V82VFIL,M46MI,which were found in 1 case,respectively.The mutations in RT region were as follows:4 cases were T69D,6 were A62V,1 was D67DE,1 was E44D,3 were V179D,1 was V179E.1 was K238KN,1 was L234T+P236S and 1 was V106E.Conclusions The major transmission route of HIV-I infection in Honghe district,Yunnan Province is through drug injection.The major HIV-1 subtype of HIV-infeeted individuals is 08_BC.PR inhibitor and RT inhibitor drug resistance associated mutations in HlV-1 gene have already existed.

In this paper, duloxetine was chosen as the lead compound. The pharmacophores with 5-HT(1A) antagonism activity were used to replace the naphthyl of duloxetine. A series of duloxetine derivatives had been designed and synthesized and whose structures were confirmed with elemental analysis, MS and H NMR. All synthesized compounds were tested by tail suspension test and forced swimming test in vivo. The test results revealed that most of the compounds have shown better activity than duloxetine at the same dosage. Some of them are worth to be studied further.

Atherosclerosis is an immune-mediated inflammatory disease of the arterial wall, with both the innate and adaptive immune systems responding to many endogenous and exogenous antigens. Both proatherogenic as well as atheroprotective roles have been identified for the immune system in atherosclerosis. Hence, it is conceivable that an immuno-modulatory strategy via active immunization against many of these antigens could potentially alter the natural history of atherosclerosis. Cholesterol ester transfer protein (CETP) plays an important role in lipid metabolism, suggesting it might prevent atherosclerosis. This review discusses the recent advances of vaccine targeting the development of atherosclerosis, mainly about the CETP targeting vaccines.

Objective:To explore the electro-clinical characteristics of Jeavons syndrome complicated with non-convulsive status epilepticus (NCSE).Methods:Three patients of Jeavons syndrome complicated with NCSE during electroencephalogram (EEG) monitoring in the EEG Monitoring Center, Department of Neurology, Xijing Hospital in 2018 were studied and followed up.Results:Among the three patients, there are two female patients, one male patient, aged 12 to 24 years. Generalized spike-wave discharges and impaired consciousness were recorded during video-EEG monitoring, which lasted for 8 minutes to 6.5 hours and evolved in generalized tonic clonic seizure (GTCS), and did not recur after receiving correct anti-epileptic drugs.Conclusion:NCSE can also occur in Jeavons syndrome, and NCSE in patients with Jeavons syndrome ends with GTCS, suggesting that early detection and timely termination of NCSE in patients with Jeavons syndrome may avoid the occurrence of GTCS.

AIM: To evaluate the safety and tolerance of ibandronate in Chinese healthy volunteers. METHODS: The trial protocol was designed according to the Good Clinical Practice(GCP). After physical examination and laboratory tests were performed, 36 healthy volunteers were divided randomly into 6 dose groups, including 1 mg , 2 mg , 3 mg , 4 mg , 5 mg and 6 mg , with 6 subjects in each group(3 male and 3 female). Clinical symptoms, vital signs, routine blood tests, routine urine tests, hepatic function, renal function, blood electrolytes, electrocardiogram, and electroencephalogram were observed or examined before and after a single intravenous infusion of ibandronate. RESULTS: After single intravenous infusion doses of 1- 6 mg , the vital signs, clinical symptoms and laboratory tests were all in the normal range, but there were some slight ADRs concerned with the drug, such as hypophosphataemia, increased body temperature, perspiring,pain of bone or muscle and hypocalcaemia. But the ADRs were found vanishing in one or two weeks. CONCLUSION: Single intravenous infusion (up to 6 mg ) of domestic ibandronate in 36 chinese healthy volunteers is safe and tolerable.

OBJECTIVE: To establish the normal value of the vestibular evoked myogenic potential (VEMP), and to determine the characteristics of VEMP in patients with acoustic neuroma (AN) and to explore the significance of VEMP in diagnosis of AN. METHOD: Click-evoked VEMP was recorded with surface electrodes attached on the sternocleidomastoid muscle. Latencies and amplitudes of specific waveform of VEMP were measured. The hearing normal subjects including 26 males and 20 females were chosen to establish the normal value of VEMP. VEMP was investigated in 14 patients with AN who underwent surgery during the period of 2006-2007 as well as auditory brainstem response (ABR) and vestibular caloric test. RESULT: Of 46 subjects with normal hearing, VEMP was present in both ears in 43 subjects, absent in either ear in three subjects. The reducible rate is 93.5% (86/92). The nor-mal value obtained from 86 reducible ears were as follows (means +/- standard deviation): latency of p13 (11.86 +/- 2.11) ms, latency of n23 (18.57 +/- 2.19) ms, interval time between p13 and n23 (6.71 +/- 1.69) ms, amplitude of p13n23 (24.18 +/- 8.22) microV. Interaural variances in 43 subjects whose VEMP were available were as follows (means +/- standard deviation): /deltap13 (0.64 +/- 0.61) ms, /deltan23/(1.05 +/- 0.97) ms, interval time between /delta13n23/ (0.84 +/- 0.81) ms, amplitude ratio (max/min) 1.32 +/- 0. 37, interaural asymmetric ratio of VEMP 0.12 +/- 0.11. Of the 14 patients with AN, VEMP was absent on the affected side in eight patients, absent on either side in three patients, and present on the unaffected side in 11 patients. VEMP presented on the affected side in three patients was significantly prolonged in /deltapl3/ and /deltap13n23/. CONCLUSION Patients with AN characterized with VEMP could be useful in the diagnosis of AN combined together with other tests.
Acoustic Stimulation ; Adult ; Case-Control Studies ; Electromyography ; Evoked Potentials, Auditory ; Female ; Humans ; Male ; Middle Aged ; Neuroma, Acoustic ; physiopathology ; Vestibular Function Tests ; Young Adult

OBJECTIVETo establish a rapid method of detecting CYP21 gene mutations.

METHODSFifty Chinese patients with 21-hydroxylase deficiency and some of their families were investigated. Blood samples were obtained for extraction of peripheral blood lymphocytes. A search for restriction sites discriminating between the morbid and the normal in CYP21 gene was made by the computer program DNAssist. PCR-based amplication-created restriction site(PCR-ACRS) was performed at I172N and R356W which are not natural recognition sequence. In addition, I172N and R356W were analysed in five families which conform to the applicability of PCR-ACRS.

RESULTSIn 50 identified 21-hydroxylase deficient Chinese patients, 21 were found to have I172 N (3 were homozygote, 18 were heterozygote); 8 were found to have R356W, all of them were heterozygote. By analysing the families, the findings were consistent with the characteristics of autosomal recessive genetic deficiency.

CONCLUSIONAnalysis of CYP21 gene point mutations using PCR-ACRS is relatively simple, accurate and feasible.

Adrenal Hyperplasia, Congenital ; enzymology ; genetics ; China ; DNA ; genetics ; metabolism ; DNA Mutational Analysis ; methods ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Female ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; Steroid 21-Hydroxylase ; genetics ; metabolism

OBJECTIVETo study the in vitro expression of 6 novel missense mutations (R270G, P275A, F121L, A156P, E183G, I324N) and a previously described R408Q mutation of phenylalanine hydroxylase (PAH) gene and explore the genotype-phenotype correlation through comparison of protein levels and residual enzyme activities.

METHODSSeven expression vectors containing PAH cDNA were constructed with a site-directed mutagenesis kit. The plasmids were extracted and sequenced to confirm the target mutations. pcDNA3.0 containing PAH cDNA was transfected into COS-7 cells and total proteins were extracted 48 h after transfection. The quantities of proteins and residual enzyme activities of the 7 mutants were assessed with the wild-type PAH gene as reference.

RESULTSRelative quantities of PAH proteins for R270G, P275A, F121L, A156P, E183G, I324N and R408Q were 10.5%, 56.6%, 54.3%, 8.7%, 8.5%, 67.3% and 85.4%, respectively. The residual enzyme activities were 7.7%, 27.6%, 19.0%, 10.4%, 9.1%, 50.6% and 40.2%, respectively.

CONCLUSIONPAH residual enzyme activities of 7 PAH mutants were all significantly reduced.

Amino Acid Sequence ; Animals ; COS Cells ; Cercopithecus aethiops ; Genetic Association Studies ; methods ; Humans ; Molecular Sequence Data ; Mutation, Missense ; Phenylalanine Hydroxylase ; genetics ; Sequence Alignment