Objective Protein N-SRCR derived from salivary agglutinin (SAG) inhibits HIV-1 infection.An N-SRCR monoclonal antibody was prepared for the study of the interaction between N-SRCR and HIV-1 envelop glycoprotein (gp120).Methods The purified recombinant N-SRCR expressed by 293 cells was used to immunize four weeks old BALB/c mice.After the final boost,the mouse spleen cells were isolated and fused with mouse myeloma cell line SP2/0-Ag-14,and the resulting hybridomas were screened for the production of N-SRCR-specific antibodies by ELISA assay.The monoclonal antibody against N-SRCR was purified by HiTrap Protein G kit,the purity determined by SDS-PAGE and the antibody titers by ELISA.The antibody specificity.was charqacterized by western blotting.Results A strain of hybridoma cell clones stably secreting N-SRCR antibody,named 1D6,was obtained.The high purity of the IgG was demonstrated by SDS-PAGE,and the ELISA titers of 1D6 was more than 100?25.Conclusion A monoclonal antibody against N-SRCR was successfully prepared,which laid the ground for further studies on the biological function of N-SRCR and the interactions between SRCR domains and HIV-1 Env gp120.

OBJECTIVE:To provide evidence for strengthening education and training before clinical practice and employment guidance among pharmacy junior students of our university. METHODS:Anonymous questionnaire survey was conducted for our university. Questionnaire survey included student employment intention and internship units,relationship of internships with employ-ment,the tendency of employment after graduation,personal attitude about pharmaceutical professional development prospect,ex-pected monthly income in the first year of their career,etc. RESULTS:Totally 71 questionnaires were sent out,71 were effectively received. Among all respondents,74.3%(52 students)believed employment intention had great relationship with internship units;24.3%(17 students)thought employment intention had not great relationship with internship units;moreover,one student thought employment intention had no any relationship with internship units. Most students(94.3%,66 students)with good attitude on in-ternships believed it could accumulate work experience;32.9%(23 students) thought they could stay in the internship unit after practice;moreover,17.1%(12 students) believed practice had little effect on employment. Among them,94.0%(63 students) tended to be employed after graduation,and only 6.0% chose a graduate school to continue their studies. Among the students who chose employment after graduation,76.1%(51 students)tended to engage in pharmaceutical related work,while 17.9%(12 stu-dents) tended to be engaged in work nothing to do with pharmacy or self-employed. The majority (69.6%,48 students) believed that pharmaceutical major prospect was general and pharmaceutical major development varied from person to person;17.4%(12 students)thought pharmaceutical major had good prospect and was promising;13.0%(9 students)believed that pharmaceutical ma-jor had no good prospect and didn't know its prospect. Most of the students(78.2%,54 students)expected a monthly income of 2500-5000 yuan in the first year of their career;18.9%expected a monthly income more than 5000 yuan(13 students);the minor-ity expected a monthly income of 1500-2500 yuan or more than 8000 yuan. CONCLUSIONS:The view of students on internship and employment have a certain gap with the social situation. Related departments of colleges and universities need to adjust the thought of students and strengthen guidance.

To characterize the background current in fetal human nasopharyngeal epithelial cells and clarify its relationship with volume activated Cl(-) currents (I(Cl,vol)), whole-cell patch clamp and cell imaging techniques were employed. Under isotonic conditions, a background current [(5.9+/-2.1) pA/pF at +80 mV, n=21] was detected. The current presented a weak outward rectification and a negligible time-dependent inactivation. The current-voltage relationship showed that the reversal potential of the background current [(-0.73+/-1.7) mV, n=21] was close to the calculated equilibrium potential for Cl(-)(-0.9 mV). Application of extracellular hypertonic stimulation (440 mOsmol/L) suppressed the current by (59.6+/-7.1)% and the inhibition was reversible after returned to isotonic conditions. Bathing the cells in hypotonic solution (160 mOsmol/L) induced a volume-sensitive Cl(-) current. The Cl(-) channel blockers, tamoxifen (20 micromol/L) and 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) (100 micromol/L), inhibited the background current by (74.0+/-5.2)% (P<0.01, n=5) and (60.9+/-8.9)% (P<0.01, n=6) at +80 mV and increased basal cell volume by (107.7+/-2.9)% (P<0.01, n=25) and (104.4+/-2.4)% (P<0.01, n=19), respectively. The data indicate that Cl(-) current is an important component of the background current in fetal human nasopharyngeal epithelial cells. The background Cl(-) current is involved in volume activated Cl(-) current and basal cell volume regulation.
Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; physiology ; Electrophysiology ; Epithelial Cells ; cytology ; metabolism ; physiology ; Fetus ; Humans ; Nasopharynx ; cytology ; Nitrobenzoates ; pharmacology ; Patch-Clamp Techniques ; Tamoxifen ; pharmacology

OBJECTIVETo investigate the effect of the rib cage on the vertebral axial rotation of adolescent idiopathic scoliosis under axial load condition.

METHODSThree dimensional finite element model of adolescent idiopathic scoliosis included and excluded thoracic cage was built based on the data of computer tomography. The model was imported into the preprocessor of the ANSYS 8.0 software for assigning boundary and loading conditions. Then the axial loading condition was simulated after entering the solution modular. The magnitude and direction of each vertebral axial rotation of the scoliotic spine were read and analyzed in the postprocessor of the ANSYS software.

RESULTSThe rib cage had a significant influence on the axial rotation of the vertebra above the structural curve and had no influence on the axial rotation of the lumbar and sacral vertebra. The effect of the thoracic cage on the axial rotation of the apical vertebra was limited. Under different loading conditions, the apical vertebra of both models rotated in the same direction. The magnitude of the vertebral rotation of both models has no statistical significance.

CONCLUSIONSAdolescent idiopathic scoliosis can lead to the anatomical changes of the vertebra and the thoracic cage. The corresponding changes of biomechanical features of the scoliotic spine and rib cage would occur. The deformed thoracic cage could not maintain the rotation stability as the normal one.

Adolescent ; Biomechanical Phenomena ; Finite Element Analysis ; Humans ; Male ; Ribs ; diagnostic imaging ; pathology ; Rotation ; Scoliosis ; diagnostic imaging ; pathology ; Spine ; diagnostic imaging ; pathology ; Thoracic Wall ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed

AIMTo explore the molecular biological mechanism of hemoglobin with high oxygen affinity in Tibetans by determining the sequence of globin cDNA in Tibetans living at high altitude.

METHODSTotal RNA was isolated from human bone marrow samples of three Tibetans who live in Qinghai-Tibet plateau. cDNA fragments coding for alpha and beta genes of human hemoglobin were obtained through RT-PCR and were ligated to plasmid pGEM-T easy vectors, and then the ligation liquid were transformed to Escherichia coli and cloned and sequenced. Nucleotide sequences were compared with GenBank data by BLAST method.

RESULTSsequence of a globin cDNA in Tibetans were the same with the registering globin genes in the GenBank, and Hb Abruzzo (beta143 (H21), His- > Arg) gene mutation, a high oxygen affinity beta globin mutation, was found in one Tibetan' beta goblin coding gene (CAC- > CGC).

CONCLUSIONThis hemoglobin gene mutation may be associated with high altitude adaptation of Tibetans living at high altitude.

Adult ; Altitude ; Asian Continental Ancestry Group ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; RNA ; Sequence Analysis, DNA ; Tibet ; alpha-Globins ; genetics ; beta-Globins ; genetics

To study the influences of warming kidney prescription on antiviral therapeutic efficacy and creatine kinase (CK) level in telbivudine-treated HBeAg-positive chronic hepatitis B patients with kidney yang deficiency syndrome. Ninety-six cases were enrolled and randomly divided into two groups (n=48 each): warming kidney prescription treatment or control. Both groups were treated for 52 weeks with telbivudine monotherapy, but the treatment group received additional treatment with the warming kidney prescription. Traditional Chinese medicine (TCM) syndrome score, biochemical response, virological response, serological response, CK level, and adverse reactions were recorded for each group in order to perform comparative analysis of the warming kidney prescription's effects. A total of 84 patients, including 43 cases in the treatment group, completed the study. The warming kidney prescription led to significantly improved total clinical syndrome efficacy, TCM syndrome score, biochemical response, virological response, and HBeAg serological responses, as evidenced by changes for each parameter observed in the treatment group versus the control group (respectively, 88.37% vs. 63.41%, 4.97+/-1.88 vs. 10.13+/-3.72, 95.35% vs. 75.61%, 81.40% vs. 56.10%, 48.84% vs. 26.83% (all, P less than 0.05)). No patient in either group experienced primary treatment failure. Seven cases, all from the control group, experienced virological breakthrough. Elevated CK was observed in both the treatment and control groups, but significantly more patients in the control group experienced this adverse reaction (respectively, 73.17% vs. 44.19%; P less than 0.01). The warming kidney prescription can increase telbivudine antiviral therapeutic efficacy and decrease the telbivudine-induced increase in creatine kinase in HbeAg-positive chronic hepatitis B patients with kidney yang deficiency syndrome.
Adult ; Antiviral Agents ; therapeutic use ; Creatine Kinase ; blood ; Female ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Male ; Middle Aged ; Nucleosides ; therapeutic use ; Phytotherapy ; Pyrimidinones ; therapeutic use ; Thymidine ; analogs & derivatives

The transwell chamber migration assay and the patch-clamp technique were used to investigate the volume-activated Cl(-) current (I(Cl.vol)) in migrated nasopharyngeal carcinoma cells (CNE-2Z). 47% hypotonic solution activated a ICl.vol in the migrated CNE-2Z cells. Compared with the control cells (non-migrated), the properties of this current and the sensitivity to Cl(-) channel blockers were changed. The current density in migrated CNE-2Z cells was higher than that in non-migrated cells. The current was almost completely inhibited by extracellular application of adenosine-5'-triphosphate (ATP, 10 mmol/L), 5-nitro-2-3-phenylpropylamino benzoic acid (NPPB, 100 mmol/L) and tamoxifen (30 mmol/L) in all voltage steps applied. The inhibition of NPPB and tamoxifen on the current was stronger in migrated cells than that in non-migrated cells. The permeability sequence of the four anions was Br(-)>Cl(-)> I (-)>Gluconate. The sequence was different from that of the non-migrated cells (I(-)> Br(-)> Cl(-)> Gluconate). The results suggest that volume-activated chloride channels may be involved in the CNE-2Z cell migration.
Carcinoma ; drug therapy ; metabolism ; pathology ; Cell Cycle ; drug effects ; physiology ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cell Size ; drug effects ; Chloride Channels ; antagonists & inhibitors ; metabolism ; physiology ; Chlorides ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; metabolism ; pathology ; Nitrobenzoates ; pharmacology ; Patch-Clamp Techniques ; Tamoxifen ; pharmacology ; Tumor Cells, Cultured

Whole-cell patch clamp and cell volume measurement techniques were used to investigate the ATP-activated chloride current and the ATP effect on cell volume in nasopharyngeal carcinoma cells. Extracellular application of ATP in micromolar concentrations activated a current with the properties of modest outward rectification and negligible time-dependent inactivation in a dose-dependent manner. The current reversed at a potential [(-0.05+/-0.03) mV] close to the Cl- equilibrium potential (-0.9 mV). Substitution of Cl- with gluconate in the extracellular solution decreased the ATP-activated current and shifted the reversal potential positively. NPPB, one of the chloride channel blockers, inhibited the current by (81.03+/-9.36)%. The current was also depressed by the P2Y purinoceptor antagonist, reactive blue 2, by (67.39+/-5.06)%. ATP (50 micromol/L) decreased the cell volume under the isotonic condition. Depletion of extracellular and intracellular Cl- abolished the ATP effect on cell volume. The results suggest that extracellular ATP of micromolar scales can induce a chloride current associated with cell volume regulation by activation of chloride channel through binding to purinoceptor P2Y.
Adenosine Triphosphate ; physiology ; Cell Size ; drug effects ; Chloride Channels ; antagonists & inhibitors ; metabolism ; physiology ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Nitrobenzoates ; pharmacology ; Patch-Clamp Techniques ; Tumor Cells, Cultured

BACKGROUNDChronic dermal ulcers are also referred to as refractory ulcers. This study was conducted to elucidate the therapeutic effect of laser on chronic dermal ulcers and the induced expression of heat shock factor 1 (HSF1) and heat shock protein 70 (HSP70) in wound tissues.

METHODSSixty patients with 84 chronic dermal ulcers were randomly divided into traditional therapy and laser therapy groups. Laser treatment was performed in addition to traditional therapy in the laser therapy group. The treatment efficacy was evaluated after three weeks. Five tissue sections of healing wounds were randomly collected along with five normal skin sections as controls. HSP70-positive cells from HSP70 immunohistochemical staining were counted and the gray scale of positive cells was measured for statistical analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to determine the mRNA and protein expressions of HSF1 and HSP70.

RESULTSThe cure rate of the wounds and the total efficacy in the laser therapy group were significantly higher than those in the traditional therapy group (P < 0.05, P < 0.01, respectively). Immunohistochemical staining revealed that the HSP70-positive cell count was significantly higher in laser therapy group than those in the traditional therapy group and controls (P < 0.01), and the gray scale of the cell signal was obviously lower than traditional therapy group and controls (P < 0.05). By contrast, the traditional therapy group and the control group were not significantly different. The RNA levels of HSF1 and HSP70 were higher in the laser therapy group by RT-PCR, but very low in normal skin and the traditional therapy group. The analysis on the gray scale of the Western blot bands indicated that the expression of HSF1 and HSP70 in the laser therapy group was significantly higher than in the traditional therapy group and the control group (P < 0.01), and the expression in the traditional therapy group was also higher than in the control group (P < 0.05).

CONCLUSIONLaser-aided therapy of chronic dermal ulcers plays a facilitating role in healing due to the mechanism of laser-activated endogenous heat shock protection in cells in wound surfaces.

Adult ; Aged ; Blotting, Western ; Chronic Disease ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Gene Expression ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Immunohistochemistry ; Laser Therapy ; methods ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Skin Ulcer ; genetics ; metabolism ; surgery ; Transcription Factors ; genetics ; metabolism

BACKGROUND:The researchers noted that upon embedding clinical-grade catgut and polyglycolide-co-lactide threads in the normal human"Zusanli"(ST 36)acupoint,the local area displayed temporal and inflammatory stimulatory effects,resulting in thread differentiation.However,the underlying mechanism behind thread involvement remains to be studied. OBJECTIVE:To investigate the expression levels of calcitonin gene-related peptide,5-hydroxytryptamine,leukotriene B4,and bradykinin at point"Zusanli"(ST 36)in rats after embedding catgut and polyglycolide-co-lactide respectively at different time points. METHODS:110 male SD rats were divided into a blank group(10 rats),a catgut embedding group(50 rats),and a polyglycolide-co-lactide embedding group(50 rats)according to the random number table method.In the blank group,no thread was embedded.In catgut embedding group and the polyglycolide-co-lactide embedding group,the thread was embedded in the left side of the ST36 acupoint once.Tissue was collected from the left side of the ST36 acupoint area 8 hours,3,7,14,and 21 days after embedding.The expression levels of calcitonin gene-related peptide and 5-hydroxytryptamine were detected by immunohistochemistry,and the contents of leukotriene B4 and bradykinin were detected by ELISA. RESULTS AND CONCLUSION:(1)Compared with the blank group,the expression of calcitonin gene-related peptide,5-hydroxytryptamine,bradykinin,and leukotriene B4 was significantly increased in the 8 hours,3,7,14,and 21 days of the catgut embedding group(P<0.05);calcitonin gene-related peptide expression was significantly increased in 8 hours,3,7,and 14 days in the polyglycolide-co-lactide embedding group(P<0.05);the expression of bradykinin was significantly increased in 8 hours,3,and 7 days(P<0.05);the expression of leukotriene B4 was significantly increased at 8 hours,3,7,14,and 21 days(P<0.05).(2)Compared with the polyglycolide-co-lactide embedding group,the expression of calcitonin gene-related peptide was increased at 7,14,21 days after thread embedding(P<0.05),and the expression of 5-hydroxytryptamine was increased at 8 hours,3,7,14 and 21 days after thread embedding(P<0.05);contents of leukotriene B4 and bradykinin in tissues were increased at 8 hours,3,14 and 21 days after embedding(P<0.05)in the catgut embedding group.(3)The results show that calcitonin gene-related peptide,5-hydroxytryptamine,leukotriene B4,and bradykinin in the acupoint region alter after catgut embedding in the ST36 of rats,as well as the alteration of calcitonin gene-related peptide,leukotriene B4,and bradykinin is found in the acupoint region after polyglycolide-co-lactide embedding in rats,which may be one of the mechanisms involved in the local time sensitive stimulus effects caused by embedding threads at acupoints.Moreover,there is a discernible difference between the two thread types.