Objective To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis(XIAP)gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer.Methods Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized,annealed,and cloned into the pSUPER vector.After identification by restriction digestion,the correct vectors were transiently transfected into SKOV3 cells,a human ovarian cancer cell line.The XIAP mRNA was detected by RT-PCR.The proteins were detected by western blot and indirect immunofluorescence staining.Flow cytometry(FCM)analysis and methyl thiazolyl tetrazolium(MTT)assay method were applied to measure cell cycle,cell growth and sensitiveness to cisplatin.Results SKOV3 cells had a high level expression of XIAP.The vector of RNA interference,which can interfere with XIAP gene was successfully constructed.After transient transfection,the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584?124,2138?65,1973?80 and 110 ?12,respectively(P=0.0334).At the same time,the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674?274,4532?107,2322?57 and 1864?78, respectively(P=0.0127).The FCM results showed that,the vector could increase the number of cells in G_1 phase compared with parent cells and compared with the cells transfected with pSUPER(P

Objective To evaluate the long term clinical effect of porcelain laminate veneer esthetic restoration for fluorosis teeth. Methods One hundred and thirty-two fluorosis teeth of 11 patients were restored with porcelain laminate veneer for esthetic purpose and the clinical effects were evaluated with modified Rygescriteria at 1.0 week, 0.5-year, 1.0-year, 1.5-year, and 2.0-year recalled time. Results After 2.0 years application, none of the veneers debonded, "A" degree rate of marginal adaptation was more than 85%, "A" degree rate of cavosurface marginal discoloration was more than 90%, "A" degree rate of secondary caries was more than 99%, "A" degree rate of post-operative sensitivity was more than 75%, and "A" degree rate of color match was more than 70%, "B" degree rate of color match was 100%. Conclusions The research indicates that the porcelain laminate veneer is a good esthetic restoration method for fluorosis teeth and the long term clinical effect is stable.

Objective:To explore the distribution characteristics of respiratory pathogens in patients with community-acquired pneumonia in Lianyungang.Methods:A total of 612 patients admitted to the second people′s Hospital of Lianyungang City because of community-acquired pneumonia (CAP) in 2019 were selected as subjects. Sputum or pharyngeal swabs were collected to extract nucleic acids, and 13-fold nucleic acids of respiratory pathogens were detected by PCR capillary electrophoresis fragment analysis. SPSS statistical software and GraphPad5.0 statistical mapping software were used for statistical analysis.Results:The physical examination rate of respiratory pathogens in the adult group was 82.0% in winter, 48.4% in spring, 28.0% in autumn, 20.0% in summer, χ 2=38.473, P=0.000. The positive rate of nucleic acid detection was significantly different in different seasons, among which the physical examination rate of respiratory pathogens in winter was the highest. The physical examination rate of respiratory pathogens in the juvenile group was 86.0% in spring, 76.2% in winter, 71.3% in summer and 66.7% in autumn, χ 2=7.946, P=0.047 . The positive rate of nucleic acid detection was calculated according to gender grouping. The comparison of nucleic acid positive rate between adult group and juvenile group in different seasons: 86.0% vs 48.4% in spring, χ 2=19.436, P=0.000; 71.3% vs 20.0% in summer, χ 2=22.180, P=0.000; 66.7% vs 28.0% in autumn, χ 2=13.485, P=0.000; 76.2% vs 82.0% in winter, χ 2=0.758, P=0.384. Except in winter, the detection rate of nucleic acid of pathogens in the juvenile group was significantly higher than that in the adult group. Conclusions:The nucleic acid detection rate and etiological distribution characteristics of respiratory pathogens are different in patients with community-acquired pneumonia in different seasons and different age groups. 13 kinds of multiple detection methods of respiratory pathogens can provide favorable laboratory data support for the diagnosis and treatment of clinical CAP patients.

The study is to design chitosan-coated pilocarpine nitrate submicro emulsion (CS-PN/SE) for the development of a novel mucoadhesive submicro emulsion, aiming to prolong the precorneal retention time and improve the ocular absorption. CS-PN/SE was fabricated in two steps: firstly, pilocarpine nitrate submicro emulsion (PN/SE) was prepared by high-speed shear with medium chain triglycerides (MCT) as oil phase and Tween 80 as the main emulsifier, and then incubated with chitosan (CS) acetic solution. The preparation process was optimized by central composite design-response surface methodology. Besides the particle size, zeta potential, entrapment efficiency and micromorphology were investigated, CS-PN/SE's precorneal residence properties and miotic effect were especially studied using New Zealand rabbits as the animal model. When CS-PN/SE was administered topically to rabbit eyes, the ocular clearance and the mean resident time (MRT) of pilocarpine nitrate were found to be dramatically improved (P < 0.05) compared with PN/SE and pilocarpine nitrate solution (PNs), since the K(CS-PN/SE) was declined to 0.006 4 +/- 0.000 3 min(-1) while MRT was prolonged up to 155.4 min. Pharmacodynamics results showed that the maximum miosis of CS-PN/SE was as high as 46.3%, while the miotic response lasted 480 min which is 255 min and 105 min longer than that of PNs and PN/SE, respectively. A larger area under the miotic percentage vs time curve (AUC) of CS-PN/SE was exhibited which is 1.6 folds and 1.2 folds as much as that of PNs and PN/SE, respectively (P < 0.05). Therefore, CS-PN/SE could enhance the duration of action and ocular bioavailability by improving the precorneal residence and ocular absorption significantly.
Absorption ; Animals ; Area Under Curve ; Biological Availability ; Chitosan ; chemistry ; Cornea ; metabolism ; Emulsions ; Microscopy, Electron, Transmission ; Miotics ; administration & dosage ; chemistry ; pharmacokinetics ; Ophthalmic Solutions ; Particle Size ; Pilocarpine ; administration & dosage ; chemistry ; pharmacokinetics ; Rabbits ; Random Allocation ; Solubility ; Tears ; metabolism

OBJECTIVETo analyze the retention rate and its influencing factors of HIV/AIDS patients by Chinese medicine (CM) maintenance treatment in the first 5 provinces in China.

METHODSKaplan-Meier method was used to analyze the retention rate of treatment in patients. Cox hazard regression model was used to assess factors that might influence the treatment time of Chinese medication.

RESULTSTotally 2,353 patients took part in this four-year study. Of them, 1,156 (49. 1%) were male, 2,344 (99. 6%) were Han nationality, 2,260 (96%) were married, 2,219 (94.3%) had junior middle-schooling or below, the average age was 41.52 +/- 8.98 years, 1,758 (74.7%) received paid blood donation, 478 (20.3%) received blood transfusion, 737 (31.3%) were absent of symptoms, 963 (40.9%) received combined treatment of CM and Western medicine. The median time within the four years was 44.84 months. The average retention rate for 1, 2, 3, 4 years of CM treatment were 86.6%, 78.4%, 72.2%, 65.6%, respectively. The results of Cox model indicated that the drop-out risk could be reduced in combined treatment of CM and Western medicine patients (HR=0. 805, P<0.01) and AIDS patients (HR=0. 769, P<0.01). The drop-out risk could be increased by the infection route of paid blood donation (HR =1. 373, P<0.01).

CONCLUSIONSThe four-year retention rate of the 2 353 patients by CM treatment in the first 5 provinces in China was 65. 6%. Route of infection, whether or not in combination of Western medicine, and staging showed influence on CM maintenance treatment time.

Acquired Immunodeficiency Syndrome ; therapy ; Adolescent ; Adult ; China ; Drugs, Chinese Herbal ; therapeutic use ; Factor Analysis, Statistical ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Patient Acceptance of Health Care ; statistics & numerical data ; Phytotherapy ; Proportional Hazards Models ; Young Adult

Objective: To explore the correlation between moxibustion sensation and distance of moxa stick and provide reference for clinical practice. Methods: A total of 16 healthy volunteers aged 18-35 years old in college were recruited and given mild moxibustion at Shousanli (LI 10), Zusanli (ST 36), Shenshu (BL 23) and Tianshu (ST 25) with moxa stick, and the occurrence and frequency of moxibustion sensation were recorded at distances of 5 cm, 4 cm, 3 cm and 2 cm. Mild moxibustion scale was used to count the score. Results: Warm was the main moxibustion sensation, burning pain and soreness decreased with the rise of distance; for the same acupoint, score of mild moxibustion scale increased with the decrease of distance; score ranged between 5.5 and 6.5 at distance 3 cm, which was the most comfortable distance for volunteers. Conclusion: The distance of 3 cm is the most comfortable distance in mild moxibustion.

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo investigate the differential expression of different types of Smads in keloids, normal scars and normal skins and its possible clinicopathological significance.

METHODSRT-PCR and Western blot methods were used to examine the expression of Smads mRNA and proteins level in 10 cases of keloid, in 10 cases of normal scar and in 10 cases of normal skin tissues and fibroblasts. Fibroblasts of keloid, normal scar and normal skin were cultured in vitro. The expression difference were compared and analyzed by t-test, there was statistical difference when P < 0.05.

RESULTSThe mRNA and protein expression of inhibitory Smad7 were significantly down regulated in keloid compared with normal scar (P < 0.05) and normal skin (P < 0.05). However, no significant difference of the mRNA and protein expression of Smad2, 3 and the protein expression of phosphorylation of Smad2, 3 in keloid, normal scar, normal skin tissues and fibroblasts.

CONCLUSIONSThe decreased expression of Smad7 in keloid might play a significant role in the increased TGF-beta1/Smads signal transduction, which can not be terminated by autologous negative feedback cycle.

Adolescent ; Adult ; Female ; Humans ; Keloid ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Signal Transduction ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult

A rapid UPLC method for simultaneous determination of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus was established. The optimal conditions of separation and detection were achieved on an HSS T3 Column (2.1 mm x 100 mm, 1.8 microm) eluted with a gradient of acetonitrile and 0.05% aqueous phosphoric acid, at a flow rate of 0.5 mL x min(-1), detected at 254 nm. The linear response ranges of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde were 0.359-3.59 mg x L(-1) (r = 0. 999 3), 2. 834-28.34 mg x L(-1) (r = 0.999 8), 0.574-5.74 mg x L(-1) (r = 0.999 8), 2.400-24.00 mg x L(-1) (r = 0.999 9), 32.57-325.7 mg x L(-1) (r = 0.999 8), respectively (n = 6). The mean recoveries (n = 9) of the five components were 96.7%-101.0%, RSD < 2.3%. The assay demonstrates that the method has adequate accuracy and selectivity to measure the concentration of protocatechuic acid, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus.
Acrolein ; analogs & derivatives ; analysis ; Chromatography, Liquid ; methods ; Cinnamates ; analysis ; Cinnamomum ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; chemistry ; Hydroxybenzoates ; analysis ; Propanols ; analysis

The concept of neurovascular units studies the interaction between neural cells and blood vessels, regards the blood-brain barrier as the core,and relates with the neurodegenerative diseases.This paper reviewed the relationship be-tween neurovascular units and neurodegenerative diseases, such as Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease,multiple sclerosis,etc.