The absorption process is an important factor in determining the bioavailability of orally administered drugs. however, the absorption mechanism of many drugs is not clear. Caco-2 cell model is the best in vitro absorption model nowadays. It is widely used in the research on drug absorption process and absorption mechanism, especially in the aspect of traditional Chinese medicine absorption, the use of Caco-2 cell model has become the hot spot recently. Mo- reover, Caco-2 cell model is also applied in the research on drug metabolism. Therefore, Caco-2 cell model will become an important method in the research of drug absorption, and will be helpful to accelerate the process of new drug screening and development.

Objective To investigate the inhibitory effect of the cytochrome P450 3A4(CYP3A4) gene expression and function in CHL-3A4 cells lines by vector-ecpressed small hairpin interfering RNA(shRNA).Methods The shRNA expression vectors targeting CYP 3A4 gene(CYP3A4Ⅰ,CYP3A4Ⅱ,CYP3A4Ⅲ) were designed and constructed.The inhibitory effect of shRNA was detected by Western blot and RT-PCR analysis.The inhibitory effect of cytotoxic of cyclophosphamide using shRNA was measured by MTT assay.Results CYP3A4Ⅲ shRNA expressing vector significantly reduced the CYP3A4 mRNA(70%) and protein expression levels(75%) of the CYP3A4 gene in CHL3A4 cells suppression of CYP3A4 gene expression by CYP3A4Ⅲ largely reversed cyclophosphamide cytotoxicity(75%) in CHL-3A4 cells.Conclusion Vector-based RNAi could suppress CYP3A4 gene expression and function,and the use of RNAi to inhibit CYP3A4 gene expression in mammalian cells was a promising new tool for the study of cytochrome P450 gene function.

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.

Objective To summarize the clinical characteristics and make genetic diagnosis in the patients with hereditary spinocerebellar ataxia type 7 (SCA7).Methods Pedigree analysis and clinical examination were performed in one family with SCA7 by clinical findings,of which retinal morphology and visual electrophysiology were available on part numbers.The polymorphic cytosine adenine guanine (CAG) repeats in the encode region of SCA7 gene were detected by combining polymerase chain reaction with deoxyribonucleic acide (DNA) sequencing on 19 familial numbers and 12 controls.Results 6 patients were identified,who manifesting cerebellar ataxia,decreased visual acuity and colour vision defect,as was pigmentary retinopathy on fundoscopy;The 6 patients had not only extinction of the electroretinogram (ERG) but also remarkably reduced amplitudes of oscillatory potentials and flash-visual evoked potentials. On normal alleles CAG repeat size ranges from 8 to 25 repeats,wherease on mutated alleles of the 6 numbers it ranges from 50 to 97 repeats.The 6 numbers were diagnosised as SCA7 patients.One asymptomatic individual of this family,who displayed a normal allele with 18 CAG repeats and another containing abnormal expantion of 56 repeats,was diagnosised as a asymptomatic carrier whose age maybe still below the age of onset.Conclusion The clinical manifestations of SCA7 are heterogeneous,and the detection of CAG repeats can provide an effective way for the gene diagnosis and the prediction of asymptomatic patients.

yggG, a Era-binding protein gene, was isolated and cloned from the E.coli genomic DNA library. Previous studies indicated that the product of yggG gene, YggG294(amino acids 1-294), strongly inhibited the growth of host bacteria and caused the death of bacteria cells. To elucidate whether Era is related to the death of bacterial cells expressed YggG294,A double promoter expression vector that can express YggG294 and Era proteins controllably in cells was constructed. Using this vector to express YggG294 and Era protein in the same E.coli cells, then analyzed the relation between YggG294 and Era. The results showed that the ratio of Era proteins to total proteins increased with the increase of induction time in E.coli cells without YggG294 expression and with little YggG294 expression;the ratio of Era proteins to total proteins seemed to be a constant level in E.coli cells overexpressing YggG294;but we could not detect any Era hydrolyzate in E.coli cells overexpressed YggG294 could not be detected. The results also showed that pre-expression of Era protein did not produce any effect on the growth inhibition of E.coli cells caused by YggG294. These results indicate that YggG294 can not hydrolyze Era protein in E.coli cells, and that YggG-Era interaction is not associated with the death of bacteria expressed YggG294. It is thus reasonable to draw a conclusion that Era is not associated with the growth inhibition of E.coli cells caused by YggG294. YggG294 inhibits the growth of bacteria by other way.

OBJECTIVETo investigate the main diseases jeopardizing the health of the iron miners and to explore the relationship between dust exposure and malignancies as well as other diseases.

METHODSA retrospective study with a cohort of 7,469 workers employed between January 1, 1972 and December 31, 1974 in Daye Iron Ore Mine Co. in Hubei Province was conducted. Standardized mortality ratio (SMR) was calculated for the main causes of death using Chinese national mortality rates for reference.

RESULTSAll subjects were followed up through December 31, 2003 with an accumulation of 199, 108.0 person years. A total of 1,752 workers died. The cumulative mortality was 23.5%. The cancers, cerebrovascular diseases, non-malignant respiratory diseases and cardiovascular diseases were main diseases that threatened workers' life span. The SMR for all subjects was a little higher than expected based on the Chinese national mortality rates. The diseases causing the significantly higher death rate were the nasopharynx cancer, liver cancer, lung cancer, pneumoconiosis and accident with SMR 1.84, 1.51, 1.83, 14.94 and 1.25 respectively. Increased mortality was observed among dust-exposed workers in the cohort. The cumulative mortality from all causes such as stomach cancer, lung cancer, nonmalignant respiratory diseases, cardiovascular diseases and accident in dust exposed workers were significantly increased compared with those in non-exposure workers with RR 1.35, 1.83, 1.61, 2.27, 1.34 and 1.69 respectively.

CONCLUSIONThe risk factors especially dust exposure affect the health and lifespan of the iron mine workers.

Adult ; Cardiovascular Diseases ; mortality ; Cause of Death ; China ; epidemiology ; Cohort Studies ; Dust ; Female ; Humans ; Iron ; Male ; Middle Aged ; Mining ; Neoplasms ; mortality ; Occupational Exposure ; adverse effects ; Respiratory Tract Diseases ; mortality ; Retrospective Studies ; Risk Factors ; Stroke ; mortality

OBJECTIVETo explore the diaphragmatic toxicity in doxorubicin (DOX)-treated rats and the related mechanisms, as well as the effects of Shengmai Injection (SMI, ) on the diaphragmatic dysfunction.

METHODSThirty Sprague-Dawley male rats were randomly divided into three groups: control, DOX-treated and DOX+SMI treated groups. DOX was given to rats in DOX and DOX+SMI groups in 6 equal doses [2.5 mg/kg, intraperitoneal injection (i.p.)], on alternate days, over a period of 2 weeks for a cumulative dose of 15 mg/kg. SMI was given to DOX+SMI rats in 12 doses (3 mL/kg, i.p.) for a period of 2 weeks before the administration of DOX and 2 weeks during the administration of DOX. The rats in the control group received equal volume of normal saline. Subsequently, the twitch and tetanic characteristics and force-frequency relationships, and the malondialdehyde (MDA) levels and the superoxide dismutase (SOD) activities, as well as the mRNA content and proteins of inducible nitric oxide synthase (iNOS) were determined.

RESULTSThe DOX-treated rats had decreased the peak twitch tension (Pt), maximal tetanic tension (P0) and force-frequency relationship as compared with the control rats (P<0.01), while the diaphragm contractility in rats treated with SMI were significantly higher than that in DOX-treated rats (P<0.01). The DOX-treated rats had increased MAD levels and decreased SOD activities (P<0.05), and SMI decreased the MDA levels and increased the SOD activities in DOX-treated rats (P<0.05). Ultrastructure of diaphragm in the DOX-treated rats revealed typical alterations including fracture of diaphragm fibers, and edema and degeneration of mitochondria; these changes were relieved by SMI treatment. The mRNA content and protein of iNOS in DOX-treated rats were remarkably higher than those in control rats (P<0.01), while SMI decreased the mRNA expression level of iNOS in DOX-treated rats (P<0.05).

CONCLUSIONSLipid peroxidation is responsible for DOX-induced diaphragm toxicity. SMI protects diaphragm muscles and their function from DOX impairment, and these beneficial effects may be somehow correlated with the decrease in expression of iNOS and lipid peroxidation.

Animals ; Biomechanical Phenomena ; drug effects ; Blotting, Western ; Diaphragm ; drug effects ; pathology ; physiology ; ultrastructure ; Doxorubicin ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation ; drug effects ; In Vitro Techniques ; Injections ; Male ; Malondialdehyde ; metabolism ; Muscle Contraction ; drug effects ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Oxidative Stress ; drug effects ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of PTEN tumor suppressor gene combined with doxycycline on telomerase activity in human mucoepidermoid carcinoma cell line.

METHODSThe wild-type PTEN tumor suppressor gene or empty vector was introduced into mucoepidermoid carcinoma cell line in vitro, then the cancer cells were treated with doxycycline. Cancer cell survival was determined by MTT assay. Telomerase activity was determined using telomerase repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).

RESULTSCompared to the control cells, cancer cells transfected with the wild-type PTEN gene showed growth inhibition and increased sensitivity to doxycycyline, and the ratio of augment of drug sensitivity was 1.65-4.75. The telomerase activity in cancer cells treared with PTEN gene transfection or doxycycline alone decreased, however, telomerase activity in combined group decreased more remarkably.

CONCLUSIONPTEN gene in combination with doxycycline has significant inhibitory effect on telomerase activity in cancer cells.

Carcinoma, Mucoepidermoid ; Cell Line ; Cell Line, Tumor ; Doxycycline ; Genes, Tumor Suppressor ; Genetic Vectors ; Humans ; PTEN Phosphohydrolase ; Telomerase ; Transfection

OBJECTIVEThe Chinese Children's Leukemia Group (CCLG)-acute lymphoblastic leukemia (ALL) 08 protocol for childhood ALL was established in 2008. This study aims to evaluate the drug-related toxicities of CCLG-ALL 08 protocol in the treatment of childhood ALL.

METHODSA total of 114 children with newly diagnosed ALL were treated with the CCLG-ALL 08 protocol. The protocol was divided into five phases: remission induction (VDLD), early reinforcement (CAM), consolidation therapy, delayed reinforcement (DIa & DIb) and maintenance treatment. Drug-related toxicities in each phase were evaluated according to the Common Terminology Criteria for Adverse Events version 4.0.

RESULTSToxicities were more frequent in phase VDLD than other treatment phases, including hepatotoxicity (87.7%), dental ulcer (20.2%), hyperglycemia (20.2%), prolonged activated partial thromboplastin time (21.1%) and decreased fibrinogen (34.2%), with the incidence rates of severe adverse events at 7%, 0, 1.3%, 0.8% and 2.7% respectively. The incidence of allergic reaction to L-ASP was significantly higher in phase DIa than in phase VDLD (28.0% vs 7.9%; P<0.01), and there were no longer any allergic reactions in 15 patients who received continuing treatment with pegaspargase instead. There was no severe arrhythmia, myocardial ischemia, decreased left ventricular function, osteonecrosis, myopathy, organ failure or treatment-related mortality.

CONCLUSIONSThe drug-related toxicities of CCLG-ALL 08 protocol are common in phase VDLD, but they are mild and reversible. There is no treatment-related mortality. The CCLG-ALL 08 protocol for childhood ALL is safe.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; Asparaginase ; adverse effects ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Remission Induction

OBJECTIVETo study the effect of mining operations on the mental health of residents living in a mining area in Hubei province.

METHODSVillagers (n = 93) living in the mining area were compared with a control group of residents (n = 101) in mental health status, and knowledge of environment and health. The mental health status of villagers was assessed using the Symptom Checklist 90 (SCL-90), State-Trait Anxiety Inventory (STAI); The knowledge of environment and health was evaluated using a self-designed questionnaire. The urine and hair samples were collected from some subjects. The lead, cadmium, arsenic, copper and zinc contents were detected as well as the total protein, NAG, d-ALA in the urine.

RESULTSThe occurrence rate of lead, cadmium, arsenic exposure symptoms was significantly higher in the exposed group than in the control group. The urine cadmium, the hair cadmium, the hair arsenic and hair lead were significantly higher in the exposed group than in the control group (P < 0.05). The positive symptom detection rate of SCL-90 in the exposed group was 8.60% compared with 0.99% of the control group. For the SCL-90, the total scale, somatization, obsessive-compulsive, interpersonal sensitivity, depression, anxiety, hostility, paranoid ideation, psychoticism, other symptom in the exposed groups were significantly higher than those in the control group (P < 0.05), indicating the status of the psychological hygiene of the exposed group was worse than the control group. The total S-AI (anxiety) score and the sex-specific value in the exposed group were higher than the control group (P < 0.05, P < 0.01), which showed that the anxiety of the exposed group was more evident. The total T-AI score and the sex-specific value in women of the exposed group were significantly higher than the control group (P < 0.01), showing that the anxiety were more significant in the exposed group, especially the women of the exposed group.

CONCLUSIONThe mental health status of the residents who living in a mining area is affected and they have a higher S-AI and T-AI scale than those living a non-mining area.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anxiety ; epidemiology ; China ; epidemiology ; Environment ; Environmental Exposure ; adverse effects ; Female ; Health Knowledge, Attitudes, Practice ; Health Status ; Humans ; Male ; Mental Health ; Metals, Heavy ; adverse effects ; Middle Aged ; Mining ; Sex Factors ; Surveys and Questionnaires ; Young Adult