BACKGROUND:Traditional total knee arthroplasty adopts medial parapatel ar approach, which induces severe trauma, requires long-term rehabilitation, and interferes the end point of quadriceps femoral tendon on superior patel ar pole. Total knee arthroplasty by subvastus approach has no impact on the knee-extension equipment, but it cannot provide sufficient exposure of surgical field and can induce damage to femoral muscle, so its application and safety need further exploration.
OBJECTIVE:To explore clinical effect of total knee arthroplasty by subvastus approach and medial parapatel ar approach.
METHODS:A total of 78 patients with 82 knees who were candidates for total knee arthroplasty were randomly divided into two groups. Treatment group (39 cases;41 knees) was given subvastus approach and control group (39 cases;41 knees) was given median parapatel ar approach. The knee function, range-of-motion of knee joint and complications after total knee arthroplasty were observed and compared.
RESULTS AND CONCLUSION:Compared with control group, the operation time in treatment group was significantly increased, while the postoperative wound drainage, straight leg raising time and walking time were obviously reduced (P<0.05). The scores of HSS before surgery and 12 weeks after surgery were not significantly different between the two groups (P>0.05), while at 1 and 6 weeks after surgery the scores in treatment group were significantly higher than those in control group (P<0.05). The range-of-motion of knee joint and MMT rank for quadriceps muscle were significantly improved in two groups after treatment (P<0.05), and those indicators in treatment group were more significant than the control group (P<0.05). The incidences of complications in treatment group and control group were 2.6%and 15.4%, respectively, with statistical y significant difference (P<0.05). The total knee arthroplasty by subvastus approach has less impact on the extensor mechanism, improves the recovery of knee function and range-of-motion of knee joint, and reduces the incidence of complications.

In the process of evolution, pathogenic Streptococcus pyogenes secretes an immunoglobulin G-degrading enzyme IdeS which can specifically cleave the hinge region of immunoglobulin G in order to escape the immune response against the host. On the one hand, IdeS can be used for IgG fingerprinting as a tool enzyme combined with mass spectrometry technology. On the other hand, IdeS can be used to treat the antibody-responsive diseases produced by autoimmunity as a therapeutic protein. In this study, the backbone of plasmid pCold was used to construct two expression vectors of recombinant protein IdeS, which were heterologously expressed in Escherichia coli Shuffle T7. After purification by affinity chromatography, the recombinant IdeS activity was detected and their activity differences between the two were compared. Among them, the yield of the recombinant IdeS containing the His6-tag at the N-terminus was 4 mg·L^-1, and the cleavage reaction with antibody IgG1 at 1∶200 (m/m) at 37 ℃ for 30 min could complete. However, the yield of the recombinant IdeS containing both the N-terminal His6 tag and the C-terminal silica affinity tag (silica bing peptide, SiBP) is 1.5 mg·L^-1, and the degradation reaction with antibody IgG1 at 1∶20 (m/m) at 37 ℃ for 30 min could reach the end. The C-terminal fusion peptide has a great influence on the yield and activity of IdeS, which is not conducive to subsequent application in drug development. Above all, the recombinant IdeS containing the His6-tag at the N-terminus expressed by this system has high activity and can fully meet the needs of antibody drug development and mapping analysis of IgG.

Objective To study the effect of assistive functional rehabilitation exercise on cardiac functioning of patients with chronic heart failure (CHF). Methods Sixty CHF patients were divided randomly into a treatment group (the rehabilitation group) and a control group, with 30 in each group. All the patients were administered routine therapy. In the treatment group, the patients were administered rehabilitation exercises with the assistance of a electric equipment made by the authors, daily for 5 days a week for a total of 3 months. The New York Heart Association (NYHA) cardiac function grading, the left ventricular ejection fraction(LVEF), the left ventricular end diastolic diameter (LVEDD) and the brain natriuretic peptide (BNP) level in plasma as well as the 6 min walking range were observed in both groups before and after treatment. Results After 3-months of treatment, the NYHA grading, LVEF, LVEDD, BNP level in plasma and 6 min walking range were all significantly improved in both groups when compared with those before the treatment, with the treatment group improved to a significantly larger extent than the control group ( p<0.05 ). Conclusion Assistive rehabilitation exercise in addition to the routine therapy can significantly help improve the cardiac function in CHF patients.

Objective To investigate the expression level and the significances of prognosis by miR-451a in nasopharyngeal carcinoma (NPC) in Guangxi. Methods The expressions of miR-451a in 89 cases of nasopharyngeal carcinoma were detected by real time RT-PCR. The relation among the expression level , the clinicopathologic features of NPC and its prognosis were analyzed. Results The expression of miR-451a were found in all of nasopharyngeal carcinoma. The expression level of miR-451a in nasopharyngeal carcinoma was negative correlated to overall survival and disease free survival (P = 0.01,P = 0.04). Conclusions miR-451a may play a key role in detection of nasopharyngeal carcinoma with poor prognosis.

Objective To explore the effect of serum deprivation on proliferation and apoptosis of cardiomyocytes and its mechanisms. Methods Human cardiomyocytes AC16 were randomly divided into serum-deprivation group (the cells were cultured in DMEM medium without fetal bovine serum for 48 h) and control group (the cells were cultured in DMEM medium containing 10% fetal bovine serum for 48 h). CCK-8 assay was performed to test the proliferation ability, and EdU imaging was conducted to detect the DNA synthesis level. Flow cytometry was performed to analyze the apoptosis level and mitochondrial membrane potential. Western blotting was conducted to detect the protein expression levels of caspase-9, Bcl-2, P53, proliferating cell nuclear antigen (PCNA), cyclin D1, β-catenin, Wnt5a, Axis inhibition protein 1 (Axin1), dishevelled 2 (Dvl2), dishevelled 3 (Dvl3) and low-density lipoprotein receptor-related protein 6 (LRP6). Results CCK-8 assay showed that the optical density value of the control group was 1.93±0.01 and that of the serum-deprivation group was 1.08±0.08, which was 55.8% of the control group, and there was significant difference between the two groups (P<0.01). DNA synthesis was inhibited by serum deprivation, and the DNA synthesis level of the serum-deprivation group was 65.6% of the control group, with significant difference found between the two groups (P<0.05). Serum deprivation could induce cardiomyocyte apoptosis; the apoptotic rate of control group was (6.34±0.47) %, and that of serum-deprivation group was (56.83±1.90) %, which was 8.9 times of the control group, and there was significant difference between the two groups (P<0.05). Serum deprivation could decrease the mitochondrial membrane potential of cardiomyocytes, while there was no significant difference between the serum-deprivation and control groups (P>0.05). Western blotting analysis showed that serum deprivation could upregulate the protein expression of caspase-9 and downregulate the protein expression of Bcl-2, P53, PCNA and cyclin D1. Serum deprivation could inhibit the protein expression of β-catenin, Wnt5a, Axin1, Dvl2, Dvl3 and LRP6 in cardiomyocytes. Conclusion Serum deprivation can inhibit proliferation and induce apoptosis of cardiomyocytes, which may be mediated by inhibition of Wnt/β-catenin signaling pathway.

Objective:To investigate the combining therapy which not only have cured effect but also can uphold and improve the NPC patient′s immunity function after radiotherrapy and chemotherapy.Method:90 cases randomly divided into 3 groups ①Local group (local injected with IL-2 +radiotherapy+chemotherapy);②General group(ivdrip with IL-2+radiotherapy+chemotherapy);③convention group(radiotherapy+chemotherapy).Checked and observed the immunity function around the immunotherapy and after the radiotherapy and chemotherapy.Result:Cellular immunity of 3 groups are lower and humoral immunity are hypetuntion than normal person.After treated with IL-2 the cellular immunity improves but there′s no great change of the humoral immunity. The immune status of the immune groups have not obvious change than before radiotherapy,at the same time,the cellular immunity of the convention group cut down and the humoral immunity doesn′t change obviously.Conclusion:①It has some effect to uphold and improve the NPC patient′s immunity function to treat with small dosage of IL-2 before radiotherapy and chemotherapy,general treatment is better than local injection;②The three therapies have not great influence on the patient′s humoral immunity.

Objective To investigate the proliferation and identification of dendritic cells(DC)de- rived from peripheral blood of patients with bladder cancer in vitro.Methods The mononuclear cells were prepared from peripheral blood of patients with bladder cancer by Ficoll-Hypaque centrifugation method,and were induced by the recombinant cytokines hGM-CSF(50 ng/ml),hlL-4(10 ng/ml)and hTNF-?(50 ng/ ml)for 2 weeks.The growth and morphology of DC were observed through the phase contrast or electron mi- croscope,and their pheuotypes were determined by flow cytometry.The capacity of DC to activate T cell-de- pendent anti-tumor immune responses was tested by MTT method.Results The DC cultured in vitro turned into suspensive growth from adhesive situation on the 6th day,then the number of DC increased con- tinuously and the cells showed the irregular morphologic appearance of DC with veiled edges on the 8th day. Flow cytometry showed that the mature DC expressed high levels of specific markers such as CD_(1a),CD_(83), CD_(86)and HLA-DR.T cells activated by DC showed strong cytotoxicity to bladder cancer cell line BIU87 with a killing rate of(48.8?3.7)%,while the killing rate of T cells which were not activated by DC was(25.7?1.5)%;the difference of the rate between them was significant(P＜0.01). Conclusions The DC can be cultured from peripheral blood of patients with bladder cancer by induction of rhGM-CSF,rhIL-4 and hT- NF-?in vitro.This may lay an experimental foundation for further research on DC vaccine.

Objective To evaluate the effectiveness of occlusive wrapping with plastic film in the delivery room to prevent heat loss and reduce the occurrence of hypothermia on admission to the Neonatal Intensive Care Unit (NICU) in very low birth weight infants.Methods A total of 209 very low birth weight infants (birth weight＜1 500 g and gestational age＜33 weeks),born in the Second Hospital of Wenzhou Medical University (Yuying Children's Hospital) and admitted to the NICU immediately after birth from June 1,2009 to December 1,2010,were randomized to the wrap group or non-wrap group.Infants randomized to the wrap group were dried and then wrapped with polyethylene film up to their necks immediately after delivery.They were then covered with warmed blankets and transported to the NICU by the neonatal team.The non-wrap group received routine care in the delivery room following the guidelines of the Neonatal Resuscitation Program.The t test and Chi-square test were used for statistical analysis.Results The mean rectal temperature on admission to the NICU was significantly higher in infants of wrap group compared with those of non-wrap group [(36.1 ± 0.8) ℃ vs (35.5 ± 1.0) ℃,t=4.707,P＜0.01],and the incidence of hypothermia was 41.0％ (43/105) in the wrap group which was lower than that in the non-wrap group (66.3％,69/104) (x2=13.547,P＜0.01).In infants ＜29 weeks of gestation,the mean rectal temperature on admission to the NICU was higher in the wrap group than in the non-wrap group [(36.1 ± 0.8) ℃ vs (35.4 ± 1.1) ℃,t=3.051,P＜0.01].In infants ≥ 29 weeks of gestation,the mean rectal temperature on admission to the NICU was higher in the wrap group than in the non wrap group [(36.0±0.9) ℃ vs (35.5±0.9) ℃,t=3.698,P＜0.01].The arterial blood gas base excess in the wrap group and non-wrap group was-(2.9±2.7) mmol/L and-(5.0±4.2) mmol/L,respectively (t=4.473,P＜0.01).The rate of endotracheal intubation was 20.0％ (21/105) in the wrap group and 27.9％ (29/104) in the non-wrap group,(x2=1.785,P=0.182).The duration of hospitalization was (49.2± 10.7) d in the wrap group and (45.8±9.1) d in the non-wrap group,and the difference was not statistically significant (t=1.152,P=0.251).Conclusions Wrapping very low birth weight infants with plastic film in the delivery room can reduce the occurrence of hypothermia.

In order to develop a protocol for clinical grade generation of dendritic cells (DCs) for cancer immunotherapy, aphereses were performed with the continuous flow cell separator and materials were derived from 10 leukemia patients that had achieved complete remission. Peripheral blood monocytes were cultured in vitro with GM-CSF, IL-4 for 6 days, then TNF-(the TNF-group) or TNF-, IL-1, IL-6, PGE2 (the cytokine mixture group) were added to promote maturation. Cell number was counted by hematology analyzer, and phenotype study (CD1a, CD14, CD83) was carried out by flow cytometry, and the function of DCs was examined by mixed lymphocyte reaction. The results showed that (0.70+/-0.13)x10(7)/mL (the TNF-alpha group) and (0.79+/-0.04)x10(7)/mL (the cytokine mixture group) DCs were generated respectively in peripheral blood obtained by leucapheresis. The phenotypes were as follows: CD1a+ (74.65+/-4.45)%, CD83+ (39.50+/-4.16)%, CD14+ (2.90+/-1.76)% in TNF-alpha group, and CD1a+ (81.86+/-5.87)%, CD83+ (81.65+/-6.36)%, CD14+ (2.46+/-1.68)% in the cytokine mixture group. It was concluded that leucapheresis may be a feasible way to provide large number of peripheral blood monocytes for DC generation, and combined administration of TNF-, IL-1, IL-6, and PGE2 may greatly promote maturity.

Objective:To investigate the expression of chemokine receptor CXCR4 in hepatocellular carcinoma tissues,hepatocellular carcinoma cell line-MHCC97,human umbilical vein endothelial cells(HUVECs)and the ascites level of CXCL12,ligand of CXCR4,so as to lay a foundation for studying the role of CXCR4 in the metastasis of hepatocellular carcinoma.Methods:The expression of CXCR4 mRNA and protein was examined by RT-PCR and Western blotting in 21 specimens of hepatocellular carcinoma tissues,MHCC97 cells,HUVECs,and 17 specimens of normal hepatic tissues.Meanwhile,the levels of CXCL12 in ascitic fluids were assayed by ELISA in 18 hepatic cancer patients.Results:The relative expression values of CXCR4 mRNA in hepatocellular carcinoma tissues,MHCC97 cells,and HVECs were 2.21?1.09,2.14?1.15 and 1.72?1.20,respectively;and those of CXCR4 protein were 1.51?0.12,1.76?0.25,and 1.89?0.24,respectively;and those of CXCR4 protein were 1.51?0.12,1.76?0.25,and 1.89?0.24,respectively.CXCR4 mRNA and protein were not detected in normal hepatic tissues.ELISA results showed that the 18 hepatocellular carcinoma samples had a CXCL12 concentration range of 783-8 364 pg/ml(median value 6 871 pg/ml)in ascitic fluids.Conclusion:CXCR4 is highly expressed in the hepatocellular carcinoma tissues and cells,which is not associated with the clinical staging of the cancer.The elevated CXCL12 level in the ascitic fluid of cancer patients indicate that CXCR4 may play an important role in the metastasis of hepatocellular carcinoma.