Objective To study surgical therapy of the serious medial collateral ligament ruptures of the knee. Methods 12 patients with acute medial collateral ligament ruptures were treated operatively.Their degrees of injury were all Ⅲdegree that superficial and deep collateral ligaments were completely ruptured.Some of them were accompanied with the injury of medial joint capsule ligament or anterior cruciate ligament or posterior cruciate ligament.The torn medial collateral ligaments were anatomically corrected by their ends with direct suture or staple internal fixation.They were followed 6 to 18 months. Results 12 patients were graded with the Lysholm score .The postoperative score (mean, 76; range, 59 to 90) was higher compared with the preoperative score (mean, 35; range, 20 to 54), P

Objective To develop a new single-tube polymerase chain reaction amplification (ST Amp) protocol for the efficient sequencing-based typing (SBT) of human leukocyte antigen DRB1(HLA-DRB1).Methods A set of 7 group-specific exonic 5′ amplification primers and a single generic 3′ primer were included together in a single PCR mix to facilitate a single PCR amplification per sample for HLA-DRB1 typing.Results All samples were successfully typed, the typing result was accurate and repeatable.Conclusion ST Amp technique has resulted in the ability to perform high-resolution, high-specificity and high-throughput HLA-DRB1 typing by DNA sequencing.

Objective To investigate the effect of multidrug resistance protein 4 (MRP4) overexpression on lipopolysaccharide (LPS)-induced vascular endothelial hyperpermeability of rat pulmonary micro-vascular endothelial cells (PMVECs) and its molecule mechanism. Methods Three to six passages of PMVECs were cultured in vitro, and they were divided into three groups: the cells in LPS group were only challenged by LPS 10 μg/mL after being cultured in serum-free medium for 24 hours; the cells in Ad-shRNA and Ad-MRP4 groups were infected with the empty virus control or recombinant adenovirus expressing MRP4 for 2 hours, and then were cultured in serum-free medium for 24 hours followed by stimulation of LPS 10 μg/mL. Endothelial permeability was assayed by the Transwell chamber models at 2, 6, 12, and 24 hours after LPS stimulation. Intracellular cyclic adenosine monophosphate (cAMP) levels were detected by enzyme-linked immunosorbent assay (ELISA). The morphological characteristics and distribution of F-actin was determined by laser confocal fluorescence microscope. The protein expressions of MRP4,β-catenin, vascular endothelium-cadherin (VE-cad) and ZO-1 were measured by Western Blot. Results ① After LPS stimulation, endothelium permeability and intracellular cAMP levels in PMVECs were significantly increased, peaked at 12 hours, and then decreased after 24 hours. Compared with LPS group and Ad-shRNA group, PMVECs of Ad-MRP4 group were exhibited a significant increase in endothelial permeability [12-hour permeability (A value):1.88±0.06 vs. 1.12±0.17, 1.10±0.18] and a significant decrease in intracellular cAMP level [12-hour cAMP (μg/L):2.39±0.02 vs. 2.97±0.01, 3.00±0.02, all P < 0.05]. There was no significant difference in endothelium permeability and intracellular cAMP levels at all time points between the LPS group and the Ad-shRNA group (all P > 0.05).② Under laser confocal fluorescence microscope, after LPS stimulation, the stress fiber formation was induced in three groups. But there were pronounced irregular aggregation of fiber in PMVECs of Ad-MRP4 group. ③ Furthermore, compared with LPS group and Ad-shRNA group, protein expression of MRP4 in Ad-MRP4 group was dramatically increased (gray value: 0.76±0.03 vs. 0.44±0.02, 0.43±0.02, both P < 0.05), and the protein expressions of β-catenin, VE-cad, and ZO-1 were significantly decreased [β-catenin (gray value): 0.14±0.03 vs. 0.23±0.04, 0.23±0.03);VE-cad (gray value): 0.21±0.01 vs. 0.34±0.02, 0.35±0.04; ZO-1 (gray value): 0.14±0.02 vs. 0.37±0.06, 0.33±0.07, all P < 0.05]. There was no significant difference in all protein expressions between the LPS group and Ad-shRNA group (all P > 0.05). Conclusion MRP4 overexpression can decrease intracellular cAMP levels, reduce intercellular junction protein expression, and then exaggerate LPS-induced vascular endothelial hyperpermeability.

ObjectiveTo establish a mouse model for genital tract infection with different serovars of Chlamydia trachomatis(Ct),and to provide a basis for understanding the relationship between Ct serovars and virulence.MethodsSix-week-old female BALB/c mice were divided into 4 groups:study group (pretreated with progesterone and inoculated with 4.0 × 107 inclusion-forming units(IFU) of Ct serovar E,F,J and K,respectively),progesterone-control group(pretreated with progesterone and inoculated with culture medium of McCoy cells),non progesterone-control group(inoculated with 4.0 × 107 IFU of Ct serovar F),blank control group receiving no treatment.At 4 and 7 days after the inoculation,the appearance of vulva was observed,vaginal swab specimens were obtained and subjected to cell culture,direct immunofluorescence assay and real-time fluorescence-based PCR for the detection of Ct.ResultsA slight inflammation of the genital tract was observed in the mice 4 days after the inoculation with Ct serovar E,F,J or K in the study group.Cell culture,direct immunofluorescence assay and real-time fluorescence-based PCR all confirmed that Ct was present in vaginal specimens from the study group,but absent from the control groups.The longest duration of infection was 21 days in 1 out of 11 mice inoculated with Ct serovar K,followed by 17 days in 1 out of 11 mice inoculated with Ct serovar J,14 days in 5 out of 11 mice with Ct serovar F and in 2 out of 11 mice with Ct serovar K.Conclusion The model for genital tract infection with different serovars of Ct can be established in mice at a right age bypretreatment with progesterone and inoculation with a certain amount of Ct.

Objective To investigate the protective effect of multidrug resistant associated protein 4 (MRP4) inhibitor on rats with sepsis-induced acute lung injury (ALI). Methods Sixty Sprague-Dawley (SD) rats were randomly divided into sham group, sepsis group and MRP4 inhibitor MK571 treatment group, with 20 rats in each group. Sepsis model was reproduced by cecal ligation and puncture operation (CLP), and the rats in sham group were only received celiotomy without ligation and puncture. Rats in MK571 treatment group were intraperitoneally injected with MRP4 inhibitor MK571 (20 mg/kg) 30 minutes before model reproduction, while rates in sham group and sepsis group were given the same amount of normal saline. Twenty-four hours later, the femoral artery blood of mice was collected, and arterial blood gas analysis was measured. Serum tumor necrosis-α (TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA). The lung tissues were collected, and the wet/dry weight ratio (W/D) was calculated. The expression of MRP4 protein in lung tissue was determined by Western Blot. Results Compared with sham group, arterial blood pH value and arterial partial pressure of oxygen (PaO2) were significantly lowered [pH value: 7.18±0.03 vs. 7.40±0.03; PaO2 (mmHg, 1 mmHg = 0.133 kPa): 63.15±6.24 vs. 98.05±2.58], while arterial partial pressure of carbon dioxide (PaCO2) was dramatically higher in the sepsis group (mmHg: 56.60±8.30 vs. 37.85±3.18), serum TNF-α level in the sepsis group was significantly increased (ng/L: 146.24±19.99 vs. 25.77±9.83), the W/D ratio of lung tissue was significantly increased (7.75±0.47 vs. 4.09±0.58), and the expression of MRP4 protein was up-regulated in the sepsis group (gray value: 0.153±0.006 vs. 0.087±0.005, all P < 0.05). Compared with the sepsis group, arterial blood pH value (7.30±0.02 vs. 7.18±0.03) and PaO2 (mmHg: 80.30±5.34 vs. 63.15±6.24) were significantly elevated in the MK571 treatment group, while PaCO2 was dramatically decreased (mmHg: 29.25±3.24 vs. 56.60±8.30), the serum level of TNF-α was significantly decreased (ng/L: 97.96±16.72 vs. 146.24±19.99), the W/D ratio of lung tissue was significantly reduced (5.89±0.51 vs. 7.75±0.47), and MRP4 protein expression was significantly down-regulated (gray value: 0.124±0.006 vs. 0.153±0.006, all P < 0.05). Conclusion MRP4 inhibitor may improve lung function in rats with sepsis-induced ALI by down-regulating MRP4 protein expression and reducing levels of inflammatory cytokines, which exerts protective effect on ALI.

In order to better fulfill the tasks of research,to turn out more quality papers,to produce outstanding results,and to further strengthen management and supervision of scientific research,the"quantitative economic management of scientific research quotas" was established in the hospital.Applying of the measure in scientific research management in the past eight years it was shown that the desired results were achieved,the academic advancement and the personnel growth were greatly promoted.

Objective To compare the efficacy and predictability of sphere-cylinder-combined LASIK and two-zone cross-cylinder LASIK for the correction of moderate and high astigmatism.Design Prospective,comparative case series.Participants 40 eyes of 35 patients with mixed astigmatism undergoing LASIK.Methods All patients were treated with Visx Star IV LASIK system.20 eyes of 19 cases were used for sphere-cylinder combined LASIK mode and 20 eyes of 16 cases for two-zone-cross-cylinder LASIK mode.All sub- jects were followed more than 6 months.Main Outcome Measures Uncorrected visual acuity (UCVA),best spectacle-corrected visual acuity (BSCVA),spherical diopter,residual astigmatism and corneal thickness.Results For the patients who received two-zone-cross-cylinder LASIK mode,the UCVA at 6 months after surgery was 0.5 or above,and 13 eyes (65.0%) were 1.0 or above. For the patients who received sphere-cylinder-combined LASIK mode,the UCVA at 6 months after surgery was 0.5 or above,and 11 eyes (55.0%) was 1.0 or above (P=0.683).The residual astigmatism for the patients received sphere-cylinder-combined LASIK mode was (1.15?1.00)D,while for the patients received cross-cylinder LASIK mode was (1.13?0.62)D(P=0.045).The remotion depth of cornea for sphere-cylinder-combined LASIK mode and cross-cylinder LASIK mode was (36.73?13.12)?m and (15.60?6.85)?m,respectively (P= 0.031).Condusion The UCVA,residual astigmatism and corneal thickness after surgery in two-zone-cross-cylinder LASIK mode were better than that in sphere-cylinder-combined LASIK mode for the correction of moderate and high astigmatism.

Objective With the purpose of further improving blood safety and providing the reference basis for a strategy suitable for large-scale routine screening of irregular antibodies in blood service,the irregular antibodies in blood samples from donors in Shanghai area were investigated.Method Irregular blood antibodies were screened in 824 072 blood donors through combined method of saline medium and microplate papain.Result 1 246 samples of blood donors were detected with irregular antibodies among the total 824 072 samples which indicate the positive rate of 1.51‰.Conclusion Screening of irregular antibodies in blood samples from donors can effectively reduce or avoid hemolytic transfusion reactions.The combined method of saline medium and microplate papain is an effective method for large-scale routine screening of irregular antibodies in blood service.

Aim To investigate the effects of classic calcium antagonists verapamil(Ver),nifedipine(Nif),diltiazem(Dil)and the novel calcium antagonist N-n-butyl haloperidol iodide(F2)which was synthesized by our lab by regulating Ca2+-independent phospholipase A2(iPLA2)on hypoxia/reoxygenation(H/R)injury of cardiac microvascular endothelial cells(CMECs)and the mechanisms.Methods The CMECs were isolated from SD neonatal rats.The H/R model was established,then cells were treated with different concentrations of calcium antagonists and F2.The content of LDH in the cell supernatant was measured by colorimetric method.The levels of IL-6 and AA in cell supernatant were measured by ELISA;and late-stage apoptosis was measured by TUNEL.The mRNA and protein expression levels of iPLA2 in CMECs were examined by real time-PCR and Western blot analysis.Results Calcium antagonists except Dil decreased the generation of LDH,IL-6 and AA in a dose-dependent manner(P<0.05),and reduced the apoptosis(P<0.05).F2 and Ver decreased the mRNA and protein expression of iPLA2 in a dose-dependent manner,while there were no such effects for Nif and Dil.Conclusions Calcium antagonists except Dil have protective effects against H/R injury.F2 and Ver protect CMECs against H/R injury partly through iPLA2.

This study investigated the changes of CD4(+) CD25(+) regulatory T cells (Tregs) in peripheral blood of patients with hepatocellular carcinoma before and after transcatheter arterial chemoembolization (TACE). The proportion of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in peripheral blood of 33 patients with hepatocellular carcinoma was determined by flow cytometry before, 1 week and 1 month after TACE. And 25 healthy volunteers served as control. One month after TACE, the patients were divided into two groups: 22 in group A, who were in stable condition or getting better; and 10 in group B, who were deteriorating. One patient died and was excluded. The results showed that the percentage of CD4(+)CD25(+) Tregs among CD4(+) T lymphocytes did not significantly change in the 33 patients 1 week after TACE as compared with that before TACE, however, the difference was significant (P<0.01) between the patients with hepatocellular carcinoma and the healthy subjects. The percentage of CD4(+) CD25(+) Tregs among CD4(+) T lymphocytes in group A 1 month after TACE was decreased significantly in comparison with that before and 1 week after TACE (P<0.01), whereas, that in group B was increased significantly 1 month after TACE (P<0.01). It was concluded that patients with hepatocellular carcinoma had a higher proportion of CD4(+)CD25(+) Tregs in peripheral blood. TACE did not significantly affect the level of CD4(+) CD25(+) Tregs within short time (such as 1 week). The proportion of CD4(+)CD25(+) Tregs in peripheral blood 1 month after TACE was related to the prognosis of hepatocellular carcinoma.
Antineoplastic Agents/administration & dosage ; Carcinoma, Hepatocellular/*immunology ; Carcinoma, Hepatocellular/therapy ; Chemoembolization, Therapeutic/*methods ; Liver Neoplasms/*immunology ; Liver Neoplasms/*therapy ; T-Lymphocytes, Regulatory/*immunology