BACKGROUND:Pathogenesis of osteoarthritis is poorly understood,however,studies have demonstrated that vascular endothelial growth factor(VEGF)Is involved in the progression of osteoarthritis.OBJECTIVE:To detect the changes of VEGF mRNA expression of synovium in rabbit osteoarthritis and to evaluate the effect of sodium hyaluronate on its expression.METHODS:Twenty four white rabbits were divided into the normal control,physiologic saline,and sodium hyaluronate groups.The unilateral anterior cruciate ligament transection(ACLT)was performed in the physiologic saline and sodium hyaluronate groups.At weeks 4 after operation,rabbits in the physiologic saline group were injected 0.3 mL physiologic saline and in the sodium hyaluronate group received 10 g/L sodium hyaluronate injection,once per week for 5 successive weeks.All the animals were sacrificed at week 10 after operation.The cartilage changes on the medial femoral condyles were graded separately VEGF express Jon of synovium was detected by using teal time polymerase chain reaction(RT-PCR).RESULTS AND CONCLUSION:The macroscopic score showed that the cartilage degeneration in the physiologic sailne group was significantly more severe than that of the normal control and sodium hyaluronate groups(P＜0 05)The expression of VEGF mRNA was obviously decreased in the physiologic saline group than that of the normal control group(P＜0 05).of which was increased in thesodium hyaluronate group,but still smaller than the normal control group(P＜0 05).The results demonstrated that the decreased VEGF expression in synovium may involved in the progression of osteoarthritis,and sodium hyaluronate has protective effect on articular cartilage by up-regulating the VEGF expression.

Objective To compare the effects of ureteroscopic lithotripsy (URL), minimally invasive percutaneous nephrolithotomy ( MPCNL) , retroperitoneal laparoscopic ureterolithotomy ( RLU) and open ureterolithotomy (UL) for the treatment of complex upper ureteral calculi. Methods The data of 281 patients with complex upper ureteral calculi from January 2005 to January 2013 were retrospectively reviewed. 48 patients of them received treatment of URL, 113 patients received MPCNL, 67 patients received RLU and other 53 patients received UL. Results Success rates of treatment at the first time were:URL 62.5% (30/48), MPCNL 92.9%(105/113),RLU 100%(67/67) and UL 100%(53/53) . The mean blood losses during the operation were:URL (9.2 ± 1.4) mL,MPCNL (72.5 ± 5.8) mL,RLU (43.1 ± 8.5) mL and UL (100.5 ± 9.2) mL. The average operation time of URL group was shorter than three other groups, and the difference was statistically significant (P< 0.05) . The average hospital stay of URL group was also shorter than three other groups ( < 0.05) . The difference was not statistically significant in complications among four groups after operation ( >0.05) . Conclusion Clinical characteristics of patients and individual require ment should be considered comprehensively before an individual treatment choice is made for the treatment of complex upper ureteral calculi.

Objective To observe the protective effect of sodium hyaluronate (Na-HA),and its effects on mRNA expression of peroxisome proliferators-activated receptor gamma (PPAR-γ) in cartilage of rabbit osteoarthritis (OA) model.Methods Forty eight white rabbits were divided into A,B,C groups randomly.Group A were normal controls,groups B and C were underwent unilateral anterior cruciate ligament transection (ACLT).The rabbits in group B were injected normal saline after ACLT;Group C rabbits received intra-articular 1% sodium hyaluronate (HA) injections 5 weeks after surgery,0.3 ml once a week.At week 11 after the surgery,all rabbits were sacrificed.The cartilage changes on the medial femoral condyles were graded.Cartilage sections were stained with safranin-O and HE,mRNA expression of PPAR-γ was detected by real time polymerase chain reaction (Real Time-PCR).Results Cartilage degeneration in group B was significantly more severe than in groups A and C.The grey value of Safranin-O of B group were higher than groups A and C.Expression of PPAR-γ mRNA in group B was higher than that in groups A and C.Conclusion NaHA has a protective effect on articular cartilage degeneration,and the inhibitory effect on PPAR-γ mRNA expression may be one of the therapeutic mechanism of Na-HA.

Objective To investigate the inhibitory effects of histidine grafted poly (β-amino es-ter) ( HGPAEs) vector-based RNA interference ( RNAi) on the expression of gene encoding myeloid differ-entiation factor 88 (MyD88) in rat liver tissues.Methods The sequence of small hairpin RNA (shRNA) was designed based on the genetic information of MyD 88.HGPAEs vector was constructed and coupled with shRNA plasmid targeting MyD88 to construct pMyD88-HGPAEs vector.Rats were divided into five groups including control group , HGPAEs treatment group , pHK-HGPAEs treatment group , shRNA treatment group and pMyD88-HGPAEs treatment group .The rats in each group were transfected with the corresponding inter-ventions through portal vein injection .Real-time PCR and Western blot assay were performed to detect the expression of MyD88 in liver tissues 3 days after transfection .Results The pMyD88-HGPAEs vector was successfully constructed .The expression of gene encoding MyD 88 was inhibited in rats from shRNA treat-ment group and pMyD88-HGPAEs treatment group (P<0.05).Significantly decreased expression of gene encoding MyD88 at mRNA and protein levels were observed in rats from pMyD 88-HGPAEs treatment group as compared with those from other groups (P<0.01).Conclusion HGPAEs vector might be used as a po-tential gene carrier .The expression of gene encoding MyD 88 in rat liver tissues could be significantly inhibi-ted through portal vein injection of pMyD 88-HGPAEs vector .This study provided evidences for further re-search on pMyD88-HGPAEs vector in a high responder model of rat orthotopic liver transplantation .

Objective To investigate the inhibitory effects of cationic polymeric liposomes (CPLs) vector-based RNA interference (RNAi) technology on the expression of rat MHCⅡ transactivator ( CⅡTA) and MHCⅡgenes .Methods According to the genetic information of CⅡTA downloaded from GenBank, three short hairpin RNA (shRNA) sequences targeting CⅡTA sequences were designed .CPLs vectors were constructed and coupled to shRNA plasmid vectors to form pCⅡTA-CPLs vectors .Six groups including control group , CPLs control group , pHK-CⅡTA control group and three pCⅡTA-CPLs groups were set up.Rat dendritic cells (DCs) were transfected in vitro.Real time PCR and flow cytometry analysis were used to detect the expression of CⅡTA and MHCⅡat mRNA and protein levels in DCs after transfec-tion.Results The pCⅡTA-CPLs vectors were successfully constructed .Compared with control groups ,the transcription level of CⅡTA and MHCⅡand the expression of MHCⅡat protein level were significantly in-hibited in all pCⅡ TA-CPLs groups ( P<0 .01 ) .The strongest inhibitory effects of pCⅡTA-CPLs on the ex-pression of CⅡTA and MHCⅡgenes were observed in the second pCⅡTA-CPLs group.There was a positive correlation between the expression of CⅡTA and MHCⅡ.Conclusion CPLs vectors were effective gene carriers.The constructed pCⅡTA-CPLs vectors significantly inhibited the in vitro expression of rat CⅡTA and MHCⅡ, which provided evidences for further investigation on pCPLs-CⅡTA vectors in vivo.

Abstract BACKGROUND: Sodium hyaluronate is an effective treatment for osteoarthritis, but the underlying mechanism remains unclear. There is evidence that abnormal expressions of matrix metaloproteinase (MMP)-1, -3 and -9 and tissue inhibitor of metaloproteinase (TIMP)-1 and -2 show great effects on osteoarthritis. OBJECTIVE: To assess the influence of intra-articular injection of sodium hyaluronate on expressions of MMPs-1, 3, 9 and tissue inhibitor of TIMPs-1, 2 in the rabbit cartilage after osteoarthritis. METHODS: Twenty-four mature New Zealand white rabbits were divided into normal control, model, and sodium hyaluronate groups. The model and sodium hyaluronate groups underwent unilateral anterior cruciate ligament transection, and rabbits in the sodium hyaluronate group received 0.3 mL of 1% sodium hyaluronate via intra-articular injection at 4 weeks after modeling, once a week for 5 weeks. At 11 weeks folowing surgery, the rabbits were kiled and the cartilage was harvested to extract total RNA. mRNA expressions of MMPs-1, 3, 9 and TIMPs-1, 2 in the cartilage were analyzed using real-time PCR for each group. RESULTS AND CONCLUSION:Compared with the model group, the range and extent of cartilage damage was reduced in the sodium hyaluronate group (P < 0.01), and Mankin scores were noticeably decreased (P < 0.05). In the cartilage, mRNA expressions of MMPs-1, 3, 9 were enhanced and mRNA expressions of TIMPs-1, 2 were down-regulated in the model group. However, the mRNA expression levels of MMPs-1, 3, 9 and TIMPs-1, 2 in the articular cartilage were not obviously changed in the sodium hyaluronate group. These results suggest that MMPs-1, 3, 9 and TIMPs-1, 2 are involved in the progression of osteoarthritis and the therapeutic mechanism of sodium hyaluronate is not realized through the down-regulation of their expressions during development of osteoarthritis. Sodium hyaluronate for treatment of osteoarthritis is a complex process and the underlying mechanisms require further investigation.

Objective To study the radiation sensitive enhancement ratio (SER) of NS398 on esophageal cancer stem cells and adherent tumor cells and analyze the radioresistance related protein expressions.Methods ECA109 esophageal cancer stem cells were cultured in serum-free medium.Expression levels of cell surface maker CD44 + and CD271 + were analyzed by flow cytometry.MTT assay was used to detect cell proliferation after the treatments with NS398 and irradiation(0,4 and 8 Gy).The sensitization effects of NS398 on the parental cells and its spheroid were evaluated by clone formation assay.Western blot assay was performed to determine protein expressions.Results Serum-free medium was successfully applied to isolate the cancer stem cells with spherical properties.CD271 + in the spheroid cells was notable higher than that in the parent cells (t =3.81,P ＜ 0.05).After irradiation,the proliferation rate of parental cells was higher than that in spheroid cells.After the combination treatment of NS398 and irradiation,SF2 value of parental cells was lower than spheroid cells(t =2.91,P ＜ 0.05)and the SER of NS398 on parental cells was greater than spheroid cells.The expressions of Bmi-1,c-Myc,β-catenin and Cyclin D1 in spheroid cells were higher than those in parental cells (t =8.09,7.90,7.50,7.15,P＜0.05).Cyclin D1 expression levels under both cell situations increased after 4 Gy irradiation (t =9.74,6.67,P ＜0.05).Compared to the 4 Gy irradiation alone group,the β-catenin and Cyclin D1 expression levels in both parental cells (t =10.15,12.12,P ＜ 0.05) and spheroid cells (t =3.23,7.45,P ＜ 0.05) decreased in the combination group.Conclusions Esophageal cancer stem cells with high level of CD271 can be isolated with serum-free medium and it is radioresistant where β-catenin and its downstream proteins may be involved.

Objective To discuss the clinical efficacy of advanced primary liver cancer under the chemother-apy of hyperthermia combined with EOF approach.Methods 75 patient samples who had advanced primary liver cancer in total were chosen and were divided into the two groups randomly.The control group containing 37 patient samples had advanced primary liver cancer which would be applied with EOF approach.The treatment group contai-ning 38 samples had advanced primary liver cancer.The treatment strategy was to apply EOF approach combined with high -frequency hyperthermia in vitro.Results The total rate of efficiency on the treatment group and control group was 73.68% vs 48.6%.The DCR rate was 92.1% versus 56.8%,1 year survival rate was 57.8% versus 45.9%and 2 years survival rate was 10.5% versus 5.4% which showed that the results of the treatment group was obviously better than that of the control group.Conclusion The method that combining the chemotherapy of hyperthermia with EOF to treat advanced primary liver cancer can obviously improve clinical efficacy and survival period.What′s better is that this approach will not increase the toxicity of chemotherapy.

ObjectiveTo investigate the effect of vascular endothelial growth factor (VEGF) on collagen Ⅱ expression in rat articular chondrocytes in vitro.MethodsChondrocytes were isolated and cultured.Then rats were divided into 4 groups:group A(control):without any intervention; group B:10 ng/ml VEGF was added; group C:10 ng/ml IL-1β was added; group D:10 ng/ml VEGF and 10 ng/ml IL-1β were added.Messenger RNA (mRNA) expression of collagen Ⅱ was detected by using real time polymerase chain reaction (real time PCR),and the protein expression level of collagen Ⅱ was detected by Western blotting.Comparisons between groups were performed by one-way ANOVA.ResultsThe collagen Ⅱ mRNA expression levels of group B (0.78+0.07),group C (0.67+0.06) and group D (0.57+0.04) were significantly lower than those of the group A (1.00±0.08),and there was significant difference between B and D,C and D.Compared with group A(0.95+0.21),the expression of collagen Ⅱ protein in group B(0.71+0.14),group C(0.60±0.11) and group D(0.31 +0.09) was significantly suppressed.The expression of collagen Ⅱ protein in group D was significantly lower than those of group B and C.ConclusionVEGF can significantly suppress the expression of collagen II in rat articular chondrocytes.VEGF may play an important role in the development of osteoarthritis.

Objective:To discuss the clinical efficacy of laparoscopic radical cystectomy in the treatment of bladder cancer after partial cystectomy.Methods:The clinical data of 30 patients who underwent laparoscopic radical cystectomy after PC in Sichuan Provincial People's Hospital from March 2016 to August 2020 were retrospectively analyzed. Including 24 males and 6 females with an average age was 62.5 (45.5-82.5)years.6 out of 30 cases underwent pelvic lymph node dissection during PC. All patients had definite pathological diagnosis for the high-grade urothelial carcinoma after PC, and the tumor staging was pT 2-3bN 0M 0.5 patients received postoperative adjuvant chemotherapy with gemcitabine and cisplatin, 6 received postoperative adjuvant radiotherapy, 13 received postoperative adjuvant radiotherapy and chemotherapy, and all patients were received maintenance intravesical instillation. Median time for local tumor recurrence after PC was 9(5-29) months, all patients had pathological diagnosis for the high-grade papillary urothelial carcinoma, cT 2-4N 0M 0 stage.The average tumor diameter was 3.5(2.5-4.5)cm, an average number of tumors was 2(1-3). Laparoscopic salvage cystectomy was performed after recurrence.General anesthesia, supine position, 5 ports were inserted through the abdominal approach. Standard pelvic lymph node dissection (PLND) was used to clean the pelvic lymph nodes. Those who had underwent PLND no longer clean the obturator and peripheral iliac vessels, but including the common iliac vessel and the bifurcation of the abdominal aorta and lymphatic tissues around the inferior vena cava, as well as the presacral lymph nodes. Results:All 30surgeries were successfully performed. The average operative time was 270(240-310)min, average estimated intraoperative blood loss was 180(50-300)ml, and there was no blood transfusion during the perioperative period.The average number of lymph nodes dissected was 18 (10-27). There were 4 cases with positive lymph nodes, of which 3 cases were positive for 2 obturator lymph nodes, and 1 case was positive for 3 obturator and external iliac lymph nodes. No serious intraoperative complications occurred.No lymphatic leakage occurred. The average drainage duration was 4(3-7) d, and postoperative hospital stays was 9(7-20)d. The postoperative pathology was invasive high-grade papillary urothelial carcinoma, and pathological TNM stage was pT 2-4aN 0-2M 0.13 patients received postoperative adjuvant chemotherapy. The average postoperative follow-up time was 23(3-31) months. There were 2 cases of pelvic recurrence and 1 case of retroperitoneal lymph node metastasis. These 3 cases received adjuvant chemotherapy and radiotherapy. Conclusions:Radical cystectomy should be the primary treatment for recurrence of bladder cancer after partial cystectomy.