Objective To optimize the condition of converting murine naive CD4+ CD25-T cells ( effector T cells,Teffs) to CD4-CD8-double negative regulatory T cells ( DN Tregs) in vitro.Methods Naive Teffs from C57BL/6 mouse were isolated with magnetic activated cell sorting( MACS)and co-cultured with DBA/2 mature dendritic cells (mDCs) with different doses of recombinant murine interleukin-2 (IL-2).The percentage of converted DN Tregs was examined by flow cytometry after 6 days.Purified DN Tregs were co-cultured with CFSE labeled Teffs.The proliferation rate of Teffs were evaluated by flow cytometry.Results Without IL-2,the percentage of CD4-CD8-T cells was 6.21％ ± 2.03％.With IL-2,the percentage was 14.77％ ± 2.15％ ( 25 ng/ml),21.29％ ± 2.68％ (50 ng/ml),43.45％ ±4.45％ (75 ng/ml),and 28.59％ ±3.05％ ( 100 ng/ml) respectively.The IL-2 concentration of 75 ng/ml signilicantly enhanced the conversion of Teffs to DN Tregs ( separately t =10.700,8.288,6.158,3.932,all P ＜ 0.05).Highly purified DN Trega significantly suppress the proliferation of Teffs in vitro.Conclusions Teffs are converted to DN Trega in vitro with the LPS-activated allogeneic mDCs and that 75 ng/ml of IL-2 is the optimal concentration for the conversion of Teffs to DN Trogs in vitro.

Objective To evaluate the clinical value of CHO-hTSHR cells in detecting thyroid stimulating antibody (TSAb). Methods The cAMP production and TSAb activity were measured and cal-culated by stimulating CHO-hTSHR cell line with IgGs of normal control group and Graves" disease ( GD )group. TSAb positive standard was set to more than the mean + 2SD of TSAb activities in control subjects.The positive percentage of TSAb activity in GD group was calculated. Results The cAMP production and TSAb activities of GD group were higher than those of normal group[( 353. 65±126. 34 ) pmol/L vs (237.21±77. 15)pmol/L, ( 149. 08±53. 26)% vs ( 100±32. 52)%, P <0. 05] . The value that higher than 165% was set to be positive for TSAb. The positive percentage of TSAb in GD group was 50% ( 14/28). Conclusion CHO-hTSHR cell line constructed by our group is suitable for detecting TSAb activity in the sera of patients with GD.

Objective To investigate the effect of triiodothyronine on the expression of alpha subunit of C protein Co (Coa) gene in cultured rat cerebral cortex neurons. Methods Primary cultured neurons of rat were prepared from the cerebral cortex at embryonic day 19. The neurons were cultured in DMEM supplemented with 15% fetal calf serum. After 7 days, the neurons were cultured in different media; DMEM supplemented with 15% fetal calf serum (group A), DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group B), 0.5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group C) , 5nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group D) and 50nmol/L T3-containing DMEM supplemented with 15% fetal calf serum stripped of thyroid hormone (group E). The neurons were cultured for another 7 days and total RNA was extracted. Goa mRNA leves were measured by competitive RT-PCR. Results As compared with group A, Coa mRNA leves in group B, D and E were low (P 0.05). Conclusion T, has dual effects on the expression of Coa gene in cultured rat cerebral cortex neurons. T3 down-regulated Coa mRNA in cultured neuron. However, in the absence of T3, Goa mRNA was also very low. Thyroid hormone may exert their action on brain development by regulating the the expression of Goa gene.

BACKGROUND:Cleft palate repair with levator veli palatini retropositioning can achieve good effects on pharyngeal function and voice quality by dissecting the muscle and rebuilding the muscle function only at the incision, without opening the mucoperiosteal flap, OBJECTIVE: To investigate the clinical effects of cleft palate repair with levator veli palatini retropositioning versus traditional surgery treatment on speech disorder correction. METHODS:Totaly 176 children with cleft palate were randomly assigned into treatment group and control group, 88 cases in each group. Patients were subjected to traditional surgery treatment in the control group and levator veli palatini retropositioning in the treatment group. RESULTS AND CONCLUSION:The postoperative speech inteligibility in the treatment group and the control group were 81.8% and 54.5%, respectively, and there was a significant difference between the two groups (P < 0.05). After cleft palate repair, the folowing indicators were al improved in the two groups, including soft palate functional length at static state and when pronouncing, and soft palate elevation angle (P < 0.05), and moreover, these indicators were better in the treatment group than the control group (P < 0.05). Scores on high nasal tone and rhinorrhea tone were decreased significantly in the two groups after repair, and the scores were significantly lower in the treatment group than the control group (P < 0.05). These findings indicate that cleft palate repair with levator veli palatini retropositioning can effectively rebuild soft palate function and perfect velopharyngeal competence in patients, which is of great significance for speech disorder correction and voice quality recovery.

Objective To evaluate the effect of infiltration anesthesia at Calot's triangle on postoperative analgesia in the patients undergoing laparoscopic cholecystectomy.Methods One hundred and forty patients,aged 18-64 yr,with 18 kg/m2 ≤ body mass index ≤ 31 kg/m2,of ASA physical status Ⅰ or Ⅱ,scheduled for elective laparoscopic cholecystectomy,were randomly divided into 2 groups (n =70 each):control group (group A) and infiltration anesthesia at Calot's triangle group (group B).In group B,1％ ropivacaine 10 ml was injected into Calot's triangle before dissection of the gallbladder,while the equal volume of normal saline was injected into Calot's triangle in group A.The patients in both groups received patient-controlled intravenous analgesia (PCIA) for 48 h starting from 10 min before the end of surgery.The VAS score was maintained below 4 during PCIA.When VAS score ≥ 4,lasting for more than 30 min,tramadol 1.5 mg/kg was injected intravenously.The consumption of physic liquor for PCIA,and requirement for tramadol were recorded.The incidence of puncture-related damage to Calot's triangle and local anesthetic intoxication and adverse effects such as nausea and vomiting within 48 h after surgery were also recorded.The first postoperative flatus time was recorded.Results Compared with group A,the consumption of physic liquor for PCIA,requirement for tramadol,and consumption of tramadol were significantly reduced,and no significant change was found in the incidence of nausea and vomiting and the first postoperative flatus time in group B.No puncture-related damage to Calot's triangle occurred in A and B groups.There was no local anesthetic intoxication in group B.Conclusion Infiltration anesthesia at Calot's triangle can optimize postoperative analgesia in the patients undergoing laparoscopic cholecystectomy.

Objective To explore the application of single-person operated mammotome minimally inva-sive surgery in treatment of benign breast lesions. Methods Clinical information of 403 patients were collected and retrospectively analyzed. 262 patients receiving ultrasound guided single-person operated mammotome mini-mally invasive surgery for benign breast masses were selected as the treatment group,141 patients undergoing open surgery for treatment of benign breast masses were selected as the control group.Operative time, introopera-tive blood loss,length of stay,postoperative healing time, as well as the occurrence of complications were com-pared between the two groups. Result Compared with the control group, the operative time was shorter, the in traoperative blood loss was less, the scar size and the postoperative healing time were shorter for the treatment group. The difference had statistical significance (P<0.05).The rate of postoperative pain, local ecchymosis, local infection and breast deformity for the treatment group was significantly lower than those of the control group (P<0.05). The difference had statistical significance (P<0.05). The recurrence rate between the two groups had no sta-tistical significance (P>0.05). Conclusions Single-person operated Mammotome minimally invasive surgery has the advantages of good operative time,good cosmetic effects, small surgical trauma, less blood loss, fast postoper-ative healing and few complications, which is worthy of clinical application.

OBJECTIVE: To study the effect of MDR1 (P-gp) and ABCG2 on the drug resistance in Hep 2 cells. METHOD: Flow cytometry was used to detect the variations of the antitumor drugs accumulation and discharging, and activity variations when MDR1 and ABCG2 inhibitors were used in Hep-2. RESULT: The accumulation and discharging of mitoxantrone was significantly higher than the control group when ABCG2 inhibitor FTC was used in Hep-2 (P<0. 05). In contrast, P-gp did not appear similar case; To the mitoxantrone and cisplatin, there was no statistical correlation about activity of Hep-2 between P-gp or ABCG2 antagonist and the control; To the doxorubicin, combining FTC and P-gp, the activity of Hep-2 was higher than the control and difference was significant (P<. 05), In contrast, FTC and P-gp did not appear similar case when used alone; To the 5-FU, when PGP used, the activity of Hep-2 was higher than that in the control and difference was significant (P<0. 05), In con- trast, FTC and FTC+P-gp did not appear similar case; To the paclitaxel, when P-gp or FTC+P-gp used, the activity of Hep-2 was higher than that in the control and difference was significant(P<0. 05). CONCLUSION ABCG2 may lead to drug resistance mainly by changing the ability of cell in accumulating and discharging chemotherapy drugs. P-gp has other way. P-gp and ABCG2 play different roles in different drug resistance.
ATP Binding Cassette Transporter, Subfamily B ; metabolism ; ATP Binding Cassette Transporter, Subfamily G, Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Humans ; Mitoxantrone ; pharmacology ; Neoplasm Proteins ; metabolism ; Paclitaxel ; pharmacology

ObjeCtive To investigate the value of protein biochip in the diagnosis of Lyme borreliosis. Methods The serum IgM and IgG antibodies against flagellin were detected by flagellin coated-and N-hydroxysuccinimide (NHS) modified-biochips in 82 patients with neuroborreliosis (NB),35 patients with erythema migrans (EM) and 44 normal controls.The results were compared with those from enzyme-linked immunosorbent assays (ELISA) and Western blot.Results The levels of both IgM and IgG antibodies were significantly higher in the NB patients than those in the normal controls (P

Objective To evaluate the curative effectiveness and safety of human chorionic gonadotropin(HCG) treatment on children with microphallic hypospadias.Methods A total of 48 consecutive children with microphalic hypospadias were enrolled in the study,and the children were randomized into the experiment group(HCG treatment) and the control group with the research randomizer.The patients in experiment group were treated with HCG prior to surgery,and the control group did not received any hormone therapy preoperatively.All children in the experiment group and the control group underwent hypospadias repair by using transverse preputial island flap (Duckett technique) urethroplasty.Penile length,diameter of glans penis,bone age,serum testosterone level,and secondary effects were recorded before and after therapy in the experiment group.Postoperative complications were assessed with respect to fistulas,urethral strictures,diverticula,meatal stenosis,and glanular dehiscence in both groups.Results (1)Mean penile length and diameter of the experiment group increased significantly by (1.08±0.47) cm (t=-5.196,P<0.05) and (0.31±0.06) cm and there was a significant difference between before and after treatment (t=-5.080,P<0.05).(2)Urethrocutaneous fistulas were observed in 8 patients in the control group compared to 2 patients in the experiment group with a statistically significant difference (χ2=4.547,P<0.05).There was a significant difference between the overall reoperation rates of control group (9 patients) and the experiment group (3 patients,χ2=4.000,P<0.05).The penile tissue of the patients in the experiment group was soft and able to be easily separated and released during the operation and the flap had more blood supply.Conclusions Pretreatment with HCG therapy prior to hypospadias repair is beneficial to children with microphallic hypospadias.Significant penile growth was seen in the children treated with HCG and there was no obvious side effect.Moreover,pretreatment with HCG is beneficial to decrease the complications and reoperation rates of hypospadias repair which proves to be effective and safe.

Objective To construct a lentiviral vector containing mir-7-3 gene and green fluorescent protein (GFP) gene,and to detect the expression of mir-7-3 gene in U251 cells.Methods The fragments containing all the mir-7-3 gene were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP,which was transfected together with the packaging plasmids into 293T cells by CaC12.The supernatant was collected,concentrated,identified,and was transfected to U251 cells of gliomas.Fluorescent microscopy was used to observe the fluorescence in the 293T cell,and real time RT-PCR was used to examine the relative contents of mir-7-3 in U251 cells.Results Electrophores was shown that the sequence of the RT-PCR product was consistent with the data of mir-7-3 by DNA sequence analysis,indicating that the mir-7-3 gene was successfully cloned,and strong green fluorescence was observed by fluorescent microscopy.The supernatant of lentivirus-transfected 293T cells effectively infected U251 cells and the relative content of mir-7-3 was observed in the transfected U251 cells.Conclusion It is concluded that the lentiviral vector containing mir-7-3 gene was constructed successfully,which provides a basis for further study of mir-7-3 function.