OBJECTIVE To construct the lentiviral vector system expressing MDR1 small interference RNA, and identify reversal efficiency of multidrug resistant phenotype in the human laryngeal cancer multidrug resistance cell lines (LSC-1/TAX). METHODS Three target sequences of oligonucleotides were selected according to MDR-1 gene sequence, the complementary DNA contained both sense and antisense strands were designed and synthesized. After the oligonucleotides were inserted into the plasmid expression system pLVTHM, the plasmid was cotransfected along with pCMV-dR8.74 and pMD2G into 293T cell lines to package lentiviral particles. Interference efficiency of the lentiviral vector system expressing MDR1 small interference RNA was determined by RT-PCR, real time PCR and Western Blot in the human laryngeal cancer multidrug resistance cell lines (LSC-1/TAX), drug resistance was measured by MTT assay after interference. RESULTS It was confirmed by digestion and sequencing that lentiviral vector had the correct structure and could express the GFP and siRNA. The functional titer of concentrated virus was more than 1?108TU/ml. The vectors expressing 3 target sequences can infect LSC-1/TAX, and the third vector has the best interference efficiency. CONCLUSION The lentiviral vector system expres-sing MDR1 siRNA has been constructed, which is necessary to reverse multidrug resistance phenotype in the human laryngeal cancer multidrug resistance cell lines

OBJECTIVETo study the effect of exogenous Ca2+ on photosynthetic parameters of Pinellia ternate and accumulations of active components under high temperature stress.

METHODThe pigment contents of P. ternata leaves, photosynthesis parameters and chlorophyll fluorescence parameters of P. ternata leaves, the contents of guanosine, adenosine and polysaccharide in P. ternata tubers were measured based on different concentrations of exogenous Ca2+ in heat stress when the plant height of P. ternata was around 10 cm.

RESULTThe contents of total chlorophyll and ratio of chlorophyll a/b were relatively higher by spaying Ca2+. Compared with the control, spaying 6 mmol x L(-1) Ca2+ significantly enhanced the net photosynthetic rate (Pn), transpiration (Tr) and stomatal limitation (L8), but reduced intercellular CO2 concentration (C) in P. ternata leaves. With the increase of Ca2+ concentration, maximal PS II efficiency (Fv/Fm), actual photosynthetic efficiency (Yield) and photochemical quenching coefficient (qP) initially increased and then decreased, however, minimal fluorescence (Fo) and non-photochemical quenching coefficient (NPQ) went down first and then went up. The contents of guanosine and polysaccharide and dry weight of P. ternata tubers showed a tendency of increase after decrease, and the content of adenosine increased with the increase of Ca2+ concentration. The content of guanosine and polysaccharide in P. ternata tubers and its dry weight reached maximum when spaying 6 mmol x L(-1) Ca2+.

CONCLUSIONWith the treatment of calcium ion, the inhibition of photosynthesis and the damage of PS II system were relieved in heat stress, which increased the production of P. ternata tubers.

Breeding ; Calcium ; pharmacology ; Chlorophyll ; metabolism ; Dose-Response Relationship, Drug ; Heat-Shock Response ; drug effects ; Organ Size ; drug effects ; Photosynthesis ; drug effects ; Pinellia ; drug effects ; growth & development ; metabolism ; physiology ; Plant Leaves ; drug effects ; growth & development ; metabolism

Animals ; Cromolyn Sodium ; administration & dosage ; Disease Models, Animal ; Intestines ; blood supply ; Mast Cells ; drug effects ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; mortality ; pathology ; p-Methoxy-N-methylphenethylamine ; administration & dosage

To elucidate the expression of WT1 in all types of leukemias and its implications for monitoring minimal residual disease in patients with acute leukemia, the peripheral blood from 55 leukemia patients and 10 normal voluteer was detected by using FQ-RT-PCR. Follow-up monitoring of WT1 expression of peripheral blood was performed for 20 patients with acute leukemia. The results showed that the expression of WT1 gene in all types of leukemias was significantly higher than that in normal control (P < 0.001). For ANLL and ALL patients, the survival time in the group of WT1 6.8 x 10(-3), (P = 0.027). Follow-up detection of the expression of WT1 in peripheral blood samples from 20 acute leukemia patients, 7 cases relapsed after complete remission has been done. In 5 of 7 relapsed patients, the expression of WT1 had obviously increased about 2 - 3 months before clinical relapse became apparent. It is concluded that the established FQ-RT-PCR method is accurate and specific. The expression of WT1 gene is relatively high in all types of leukemias compared with normal peripheral blood cells, the higher WT1 expression may associate with poor prognosis in acute leukemia, and the dynamics of WT1 level correlate with the disease status. The quantitative assessment of WT1 expression in peripheral blood samples by FQ-RT-PCR may be a useful tool for monitoring minimal residual disease.
Acute Disease ; Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; genetics ; Child ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia ; blood ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm, Residual ; blood ; diagnosis ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; WT1 Proteins ; genetics

OBJECTIVETo investigate the effects of Huchang Qingfei concentrated pellets on the expression of E-cadherin (E-cd) in the lung tissue from mice infected with Mycoplasma pneumoniae (MP).

METHODA mice model of Mycoplasmal pneumonia (MPP) was developed by repeatedly intranasal infectious route. Transmission electronic microscope (TEM) and immunohistochemistry stain were performed to observe the pathological changes and expression of E-cd in lung tissues.

RESULTUnder TEM it was found that the cellular membrane was ruptured, mitochondria was denatured, crista was broken in the pulmonary cells of the model group; the all above parameters in Huchang medicated group were improved obviously. The immunohistochemistry test showed that strong positive brown stain of E-cd expression was found in the pulmonary epithelial cell membrane and bronchial periphery in the model group, however, in the medicated group, the E-cd expression level in the cellular membrane was decreased and the expression ratio was dropped significantly as compared with the model controls.

CONCLUSIONHuchang Qingfei concentrated pellets can inhibit the overexpression of E-cd in the lung tissue of mice with MP-infection, which may be helpful for prevention and treatment of pulmonary injury caused by MPP.

Animals ; Cadherins ; metabolism ; Cell Membrane ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Epithelial Cells ; metabolism ; Female ; Lung ; metabolism ; pathology ; Male ; Mice ; Mycoplasma pneumoniae ; isolation & purification ; Plants, Medicinal ; chemistry ; Pneumonia, Mycoplasma ; metabolism ; microbiology ; pathology ; Random Allocation

PURPOSE: This study assessed marginal bone remodeling and soft tissue esthetics after the loading of single bone-level implants in the anterior maxilla. METHODS: An open, single-arm observational clinical trial with 3 years of follow-up was performed, including 22 implants. The patients presented with a single tooth gap in the anterior maxilla (tooth positions 14–24), with natural or restored adjacent teeth. An implant was placed at least 8 weeks post-extraction and healed submerged for 6 weeks. After the second-stage operation, a fixed provisional prosthesis was provided. The final restoration was placed 6 months after the provisional restoration. The time of the provisional crown connection was considered to be the baseline in this study. Esthetic parameters and the marginal bone level were assessed at 6, 12, 24, and 36 months. RESULTS: All implants were well integrated in the bone. A statistically significant increase was found in the mean implant stability quotient between the time of the provisional prosthesis and the time of the final prosthesis. Most implants (95.5%) revealed marginal bone resorption (<0.5 mm), and just 1 implant (4.5%) showed a change of 2.12 mm from baseline to 36 months (mean 0.07±0.48 mm), while the crestal bone level decreased significantly, from 2.34±0.93 mm at baseline to 1.70±1.10 mm at 36 months. The facial gingival margin and papilla were stable and the esthetic scores indicated high patient and dentist satisfaction. CONCLUSIONS: Platform-switching bone-level implants placed in maxillary single-tooth gaps resulted in successful osseointegration with minimal marginal bone resorption. The peri-implant soft tissue was also esthetically satisfying and stable.
Alveolar Bone Loss ; Bone Remodeling ; Bone Resorption ; Crowns ; Dental Implants ; Dentists ; Esthetics ; Follow-Up Studies ; Humans ; Maxilla* ; Observational Study* ; Osseointegration ; Prostheses and Implants ; Tooth

OBJECTIVES: To test the feasibility of submandibular salivary gland (SMG) replantation techniques and the survival of the replanted glands. Such a study can provide a rationale for later allotransplantation procedures, along with implementation of conventional and advanced immunosuppression therapy. MATERIALS AND METHODS: Six SMG replantations were performed in New Zealand white rabbits. One week postoperatively, (99m)Tc scintigraphy was performed and the uptake ratio and salivary excretion fraction were calculated. Two to four weeks later, submandibular glands were excised, fixed, and stained with H&E for histomorphometric evaluation. RESULTS: Intraoperatively, all glands showed patent blood perfusion except gland 5. Positive tracer uptake and saliva excretion were documented by scintigraphy. On excision, all of the glands except glands 4 and 5 looked viable, with a red color and patent pedicles. Gland 4 was infected and filled with creamy pus, while gland 5 looked pale and necrotic. Histologically, glands 1, 2, 3, and 6 had preserved normal glandular tissue with slight variations from the contralateral normal glands, as their parenchyma was composed of mildly atrophic acini. CONCLUSION: Four out of six replanted SMGs successfully survived. The glands maintained good viability and function. Such success depends on safe harvesting, short anastomosis time, and strict control of infection.
Immunosuppression ; Perfusion ; Rabbits ; Radionuclide Imaging ; Replantation* ; Saliva ; Salivary Elimination ; Salivary Glands* ; Submandibular Gland ; Suppuration

OBJECTIVES: To test the feasibility of submandibular salivary gland (SMG) replantation techniques and the survival of the replanted glands. Such a study can provide a rationale for later allotransplantation procedures, along with implementation of conventional and advanced immunosuppression therapy. MATERIALS AND METHODS: Six SMG replantations were performed in New Zealand white rabbits. One week postoperatively, (99m)Tc scintigraphy was performed and the uptake ratio and salivary excretion fraction were calculated. Two to four weeks later, submandibular glands were excised, fixed, and stained with H&E for histomorphometric evaluation. RESULTS: Intraoperatively, all glands showed patent blood perfusion except gland 5. Positive tracer uptake and saliva excretion were documented by scintigraphy. On excision, all of the glands except glands 4 and 5 looked viable, with a red color and patent pedicles. Gland 4 was infected and filled with creamy pus, while gland 5 looked pale and necrotic. Histologically, glands 1, 2, 3, and 6 had preserved normal glandular tissue with slight variations from the contralateral normal glands, as their parenchyma was composed of mildly atrophic acini. CONCLUSION: Four out of six replanted SMGs successfully survived. The glands maintained good viability and function. Such success depends on safe harvesting, short anastomosis time, and strict control of infection.
Immunosuppression ; Perfusion ; Rabbits ; Radionuclide Imaging ; Replantation* ; Saliva ; Salivary Elimination ; Salivary Glands* ; Submandibular Gland ; Suppuration

OBJECTIVETo study whether plasma viral load testing is helpful to exclude ones free from Human immunodeficiency virus (HIV) infections from suspects in HIV antibody detections.

METHODS19 Specimens, which showed disconcordant results of the two HIV EIA testing (S/CO < 6) and indeterminated results of Western blot (WB) test, were selected. Viral load of the specimens were detected. A six-month follow up survey in detecting HIV antibody was conducted in these subjects.

RESULTSNone of these 19 cases was observed to be positive HIV viral loads and there was no any progress in WB bands development during the follow-up period. The possibility of HIV infection could be excluded.

CONCLUSIONWhen the specimens react with very low intensity in both EIA and WB, negative viral load result is conducive to exclude negative subjects from suspects in HIV antibody detections.

AIDS Serodiagnosis ; HIV Antibodies ; blood ; HIV Infections ; blood ; diagnosis ; Humans ; Viral Load

OBJECTIVETo identify a cost-efficient alternative antibody testing strategy for screening and confirmation of HIV infection by rapid simple tests (RSTs) and enzyme-linked immunosorbent assays (ELISAs).

METHODSFour RSTs (RST1, RST2, RST3, and RST4) and five ELISAs (ELISA1, ELISA2, ELISA3, ELISA4, and ELISA5) were evaluated in two phases by using banked and serum specimens prospectively collected at regional hospitals and voluntary counseling and testing (VCT) centers in Beijing. A total of 200 banked serum specimens were included in the first phase, including 62 HIV-positive, 127 HIV-negative and 11 indeterminate specimens. All specimens were tested by four RSTs and five ELISAs respectively. The second phase involved prospective testing of 389 routine specimens, including 92 HIV-positive, 287 HIV-negative, and 10 indeterminate specimens. All the specimens were tested by two RSTs (RST2 and RST4) and three ELISAs (ELISA1, ELISA3, and ELISA4), which were selected for their respective excellent sensitivity and/or specificity. Western blot (WB) was used as a gold standard for confirming the reactivity of all the specimens.

RESULTSSensitivity, specificity, and efficacy were calculated for each assay in two phases. In the first phase, four assays (ELISA4, RST2, RST3, and RST4) had a specificity of 100%. For the determination of efficacy, ELISA4, RST2, and RST4 were selected in the second phase. ELISA1 and ELISA3 which have a sensitivity of 95.9% and 93.2% respectively also entered this phase. In the second phase, all the five assays (ELISA1, ELISA3, ELISA4, RST2, and RST4) had a sensitivity and specifity of over 90%. ELISA1 had a sensitivity of 99% and ELISA4 a specificity of 99%.

CONCLUSIONThe sensitivity ELISA1 and the specificit of ELISA4 are comparable to ELISA/WB standard strategy. Application of this alternative testing strategy provides a cost-effective method for determining HIV prevalence in Beijing.

Blotting, Western ; methods ; China ; Enzyme-Linked Immunosorbent Assay ; methods ; HIV Infections ; diagnosis ; Humans ; Sensitivity and Specificity