Objective To analyze the efficacy and safety of CT-guided percutaneous injection of the fibrin glue bv double needle technique to treat sacral cyst.Methods Clinical data of 20 cases with double-needle injection of fibrin glue technology to treat sacral cyst were retrospectively analyzed.All patients had varying degrees of sacral nerve root compression symptoms.The treatment for sacral cyst was carried out after clear diagnosis was made.On the basis of CT-guided percutaneous injection of fibrin glue,the improved CT-guided percutaneous injection of fibrin glue by double-needle technique was used to treat these patients.The average dose of fibrin glue was(5.9 ± 2.4)ml.The clinical results of improvement as to pain and neurological function were evaluated after follow-up of an average of 17 months.The assessment criteria were as follows:excellent,complete resolution of signs and symptoms,with the patient returning to his or her regular employment and no recurrence of cysts during 1 year of follow-up,good,symptoms and signs in the legs and perineal region resolved but with persistent pain in the lumbosacral region,which did not interfere with the patient' s regular work (the cysts did not recur for 6 months during follow-up),fair,no improvement in clinical symptoms,but a decrease in cyst size on the imaging study,poor,no improvement in clinical symptoms and no observed changes in cyst size in imaging studies or recurrence.Results Most patients experienced some degree of pain relief and functional improvement after fibrin glue therapy,with most experiencing complete or marked resolution of clinical symptoms.Nine patients reported excellent recovery,8 reported good recovery,2 reported fair recovery,and 1 reported poor recovery.The overall percentage of positive outcomes (excellent and good recovery) was 85％.No serious postoperative complications were discovered.Conclusions CT guided percutaneous injection of the fibrin glue bydouble needle technique to treat sacral cyst is an ideal method.Double needle technique is simple,safe and reliable.

Objective To observe the expression and mechanism of monocyte chemoattractant protein-1 (MCP-1) in interstitial cystitis (IC) rats.Methods Twenty weight of 250-300 g of female SD rats were divided into IC group (n =10) and control group (n =10).IC group were treated by transurethral instillation with 10 mg/ml protamine sulfate (PS) 1 ml reserved for 45 min,and then instillation with 750 ug/ml of lipopolysaccharide (LPS) 1 ml reserved for 30 min.The same operations were repeated after 24 hours,and the rats were killed obtaining the bladder tissue and urine after three days.Control group was given PBS solution perfusion.MCP-1 and histamine (HA) expression levels in the rat bladder tissue and urine were detected by ELISA.The inflammation of bladder tissue was observed and inflammatory score was used by HE staining.MC degranulation count was used by MC special staining.MCP-1 expression and distribution in bladder tissue was observed by immunohistochemical method.The relationship between the MCP-1 and MC was detected by immunofluorescence method.Results By ELISA,the expression levels of MCP-1 and HA in the bladder tissue and urine in IC group were significantly increased compared with control group (P ＜0.01 ).More inflammatory cell infiltration in the bladder mucosa,edema mucosa,congestion and hemorrhage were seen by HE staining.The inflammatory score in IC and control group were (76.5 ±9.8) and (18.5± 9.8)/field (P ＜ 0.01 ).With MC special staining,degranulation MC count in IC and control group were (6.4±3.1 ) and (0.7 ±0.3)/field (P ＜0.01 ),and the degranulation in the bladder tissue of IC group was significantly higher than control group (P ＜ 0.01 ).MCP-1 has a higher expression in the bladder epithelium,and more MCP-1 were found gathering round MC surface by immunofluorescence.Conclusions MCP-1 is highly expressed in IC rats,and could induced activation of MC,which could release HA,aggravating the pathological process of inflammatory and fibrosis in IC.

Objective To investigate the effect of acupunctures variable with stages on lumbar intervertebral dise protrusion (LIDP). Methods 98 LIDP patients in our hospital from October, 2014 to July, 2015 were randomly divided into control group (n=49) and observa-tion group (n=49) according to the sequence of first diagnosis time. The control group received routine acupuncture, and the observation group acupunctured at Ouch point and Huantiao point (GB30) with different acupuncture according to manifestations and courses. Modified Yang acupuncture was adopted in acute phase, triple acupuncture was used in remittent phase, and lateral needling was used in the recovery phase. They were both treated once a day, 7 days as a course for 3 courses with 2 days of interval. Visual Analogue Scale (VAS), Oswestry Disability Index (ODI) and clinical effects were assessed before and 1, 2, 3 courses after treatment. Results Both VAS and ODI scores im-proved after treatment (F>7.12, P<0.05), especially in the observation group (t>4.43, P<0.05). The efficiency was higher in the observation group than in the control group (χ2=5.594, P<0.05). Conclusion Compared with normal acupuncture, the acupunctures variable with stages is more effective on LIDP than routine.

Objective To investigate the temperature variation within intra-spinal canal and intra-spinal tumor during percutaneous radiofrequency ablation (RFA) procedure for vertebral tumor in experimental rabbits. Methods Eight New Zealand white rabbits were transplanted with VX2 carcinoma in the lumbar vertebral body by percutaneous puncture inoculation technique under CT guidance in order to set up vertebral tumor models. The eight vertebral tumor models were treated with RFA under CT guidance. The temperature within the spinal canal and vertebral tumor of rabbits was measured and recorded every 30 seconds during the RFA treatment. The results were statistically analyzed by paired sampled t text. Results The intra-tumor temperature rose to 90℃ rapidly and remained stable during the whole RFA procedure, whereas the temperature in the spinal canal exceeded 42℃ when treatment time was over three minutes during the procedure. Statistically significant difference in the temperature level during RFA existed between the spinal canal and the vertebral tumor (P < 0.05). Conclusion The temperature in the vertebral tumor of rabbit can quickly reach to the therapeutic level during RFA. Prolonging operative time of RFA may hurt the nerve due to high temperature.

BACKGROUND:Soft tissue engineering mainly includes seed cells,scaffolds,cytokines and bioreactors,among which,the scaffolds are the key link in the construction of tissue-engineered cartilage.OBJECTIVE:To prepare an articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold,and to evaluate its physicochemical properties and biocompatibility.METHODS:The articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold was prepared by freeze thawing drying method using porcine articular cartilage extracellular matrix and human umbilical cord Wharton glue as raw materials.The porosity,water absorption,tissue composition and longitudinal compressive elastic modulus of the scaffold were measured and histologically stained.Rabbit chondrocytes were co-cultured with the articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold for 7 days.Then,scanning electron microscopy,live-dead cell staining and hematoxylin-eosin staining were performed.In addition,rabbit chondrocytes were cultured in the extract of the articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffold and cell culture medium for 6 days,respectively;and MTT assay was used to detect cell proliferation.RESULTS AND CONCLUSION:The articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffolds had a cross-section of uniform porous network structure and a vertical cross-section of the vertical tubular structure,and the pore wall was densely covered with cartilage fibers.The composite porous scaffold was positive for hematoxylin-eosin staining,safranin O staining and toluidine blue staining,and contained collagen and glycosaminoglycan ingredients.The water absorption,porosity and longitudinal compressive elastic modulus of the scaffolds were (17.418 8±0.909 0)％,(81.495 1±6.621 0)％ and (2.833 3±0.456 4) kPa,respectively.After 7 days of co-culture,rabbit chondrocytes adhered to the scaffold and proliferated,and further grew into the pores of the scaffold.Moreover,the scaffold was non-toxic to the rabbit chondrocytes.To conclude,the physiochemical properties and biochemical components of articular cartilage extracellular matrix/human umbilical cord Wharton gel porous scaffolds are similar to those of natural cartilage,and the scaffold has good biocompatibility.

Objective To preliminarily evaluate the safety and efficacy of percutaneous vertebroplasty (PVP) with 131I-loaded bone cement in treating vertebral tumor in rabbit models. Methods Twelve New Zealand white rabbits with lumbar vertebral tumor, which was established by puncturing transplant of VX2 carcinoma, were randomly and equally divided into the study group and the control group with 6 rabbits in each group. PVP with injection of 131I-loaded bone cement was carried out in the rabbits of the study group, while PVP with injection of pure bone cement was employed in the rabbits of the control group. The blood cell count was determined in all the animals one day before PVP as well as on the 4th day after PVP. PET-CT examination was performed one day before PVP as well as on the 4th day after PVP to check the stand uptake value (SUV) of each vertebral tumor. SPECT was performed in all rabbits of the study group at one, 4 and 8 days after PVP respectively to estimate the distribution of 131I in the animals’ bodies. Eight days after PVP, blood cell counts, which were determined both before and after PVP, existed between the study group and the control group. SPECT that was performed after PVP indicated that 131I was mainly accumulated within PVP-treated vertebrae, and the distribution of 131I showed no obvious changes at different points of time after the procedure. Before PVP, the difference in SUV between the two groups was of no statistical significance (F = 0.765, P > 0.05). In the study group, the postoperative SUV was significantly lower than the preoperative SUV (F = 423.792, P < 0.05). Pathological examination showed that the extent of tumor cell necrosis around the bone cement in the study group was remarkably bigger than that in the control group. Conclusion In treating vertebral tumors with PVP, the use of 131I-loaded bone cement is clinically feasible, and short-term follow-up indicates that this technique is safe and effective.

Ranpirnase (onconase, ONC) is a new drug, with weak RNase activity and strong cytotoxicity to various tumor cells in vitro and in vivo. This study is to obtain recombination onconase (rONC) with high bioactivity. Based on the codon preference of Pichia pastoris, we designed and synthesized the gene according to cDNA sequences of ONC and the α mating factor's prepeptide. We screened positive clones after transforming the recombination plasmids into P. pastoris X-33, GSS115 and SMD1168. We screened the best combination of seven different vectors and host strains. Moreover, we optimized culture condition in shake flasks and 10 L bioreactor, and purified rONC from the supernatant after inducing it with 0.25% methanol by aqueous two-phase extraction coupling G50 molecular exclusion method. The highest rONC production was 13 mg/L in pPICZα-A/X-33/ONC combination under the condition of pH 5.5 and 23 degrees C in shake flasks for 7 d; and that the highest rONC production was 180 mg/L when the induction is performed in the lower basic salt medium with pH 5.5 in the 10 L bioreactor for 7 d. The yield of rONC is more than 90% at a purity of above 95%. rONC can kill various tumor cells in vitro. The expression and purification of rONC would be useful for further investigation of this new drug.
Antineoplastic Agents ; metabolism ; Bioreactors ; Cell Line, Tumor ; Codon ; DNA, Complementary ; Genetic Vectors ; Humans ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; Ribonucleases ; biosynthesis

Objective To evaluate the feasibility and outcomes of robotic-assisted laparoscopic pyeloplasty in children .Methods A retrospective study was performed in patients who underwent robotic-assisted laparoscopic pyeloplasty ( Anderson-Hynes ) at our institution between January 2014 to August 2014.Totally 6 boys were diagnosed as left ureteropelvic junction obstruction depending on the symptoms and radiographic studies .The mean age was 9 years ( range 4 -12 years ) .Results The procedure was performed successfully without conversion to open surgery in all of the cases .Mean operative time was 216 min (range 175-269 min), with a mean robotic anastomosis time of 45 min (range 30-60 min).Mean estimated blood loss was less than 15 ml.The mean hospitalization was 4.5 days.Mean follow-up period was 10 months ( range 7 -14 months ) .There were no perioperative complications , and recovery was uncomplicated (without recurrence, pyelonephritis, nephrarctia) in all of the patients.Conclusion Robotic-assisted laparoscopic pyeloplasty can be safely performed in children older than 4-year-old with ureteropelvic junction obstruction .

Objective:To explore the regulation effects of baicalin on the behavior as well as extracellular regulated protein kinase(ERK)and cAMP-response element binding protein(CREB) in chronic unpredictable mild stimulus(CUMS) model mice.Methods:Thirty ICR mice were randomly assigned to control(CON) group, model(CUMS) group, fluoxetine(FLU) group, baicalin high-dose(BA-H) group and baicalin low-dose(BA-L) group with 6 mice in each group.In addition to the CON group, the mice in the other four groups were modeled by CUMS method.The modeling was carried out for 42 days, and intragastric administration was carried out according to groups from the 21st day to the completion of modeling.After administration, the depression like behavior of mice was measured by sugar water preference test and water maze test.Western blot (WB) and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the protein level and mRNA level of ERK and CREB in mouse hippocampus respectively.SPSS 21.0 was used for statistical analysis.After normal test and variance homogeneity test, one-way ANOVA was used for multi group comparison, and Tukey test was used for pairwise comparison.Results:Results from the sugar preference experiment showed that compared with CON group, the sugar preference rate of CUMS group was decreased ((82.88±2.00)%, (64.49±1.24)%, t=19.11, P<0.05). Compared with CUMS group, sugar preference rate in FLU group ((81.90±1.19) %), BA-H group (77.86±2.51)%) and BA-L group ((67.98±2.56)%) increased ( t=24.83, 11.68, 3.00, all P<0.05). The results of water maze test showed that compared with CON group, the number of crossing platform ((6.33±0.82), (1.83±0.75), t=9.93, P<0.05) and the target quadrant residence time ((46.83±4.78)s, (24.25±6.12)s, t=7.13, P<0.05) of mice in CUMS group were decreased, but the the escape latency was prolonged ((14.88±3.00) s, (70.70±4.77) s, t=24.26, P<0.05). Compared with CUMS group, the number of crossing platform ((5.00±0.89)times, (5.17±0.75)times and (3.33±0.82) times, t=6.64, 7.67, 3.31, all P<0.05), and the residence time in the target quadrant ((36.80±2.66) s, (36.82±5.62) s, (33.28±3.56) s, t=4.61, 3.71, 3.13, all P<0.05) in FLU group, BA-H group and BA-L group increased, but the escape latencies were shortened ((23.37±4.86) s, (34.83±4.72) s, (62.15±5.30) s, t=17.02, 13.10, 2.94, all P<0.05). WB results showed that compared with CON group, the expression of ERK protein ((1.00±0.15), (0.36±0.10), t= 6.26, P<0.05) and CREB protein((1.00±0.12), (0.29±0.03), t=10.32, P<0.05) in hippocampus of mice in CUMS group decreased.Compared with CUMS group, ERK protein in hippocampus of mice in FLU, BA-H and BA-L groups increased ((0.87±0.05), (0.77±0.08), (0.67±0.03), t=8.25, 5.7, 5.39, all P<0.05), and CREB protein in FLU, BA-H and BA-L groups were also increased ((0.90±0.12), (0.84±0.14), (0.62±0.04), t=8.94, 6.59, 12.25, all P<0.05). qPCR results showed that compared with CON group, ERK mRNA ((1.00±0.03), (0.41±0.10), t=9.78, P<0.05) and CREB mRNA ((1.00±0.08), (0.61±0.12), t=4.62, P<0.05) were decreased in CUMS group.Compared with CUMS group, ERK mRNA in hippocampus of mice in FLU, BA-H and BA-L groups were increased ((0.71±0.08), (0.69±0.03), (0.59±0.04), t=4.15, 4.65, 2.84, all P<0.05), CREB mRNA in FLU group and BA-H group were increased ((0.87±0.08), (0.86±0.07), t=3.14, 3.19, all P<0.05). Conclusion:BA can improve the depression-like behavior of CUMS model mice.The mechanism of action may be related to the regulation of ERK and CREB proteins.

Human fibroblast growth factor 21 (hFGF21) has become a candidate drug for regulating blood glucose and lipid metabolism. The poor stability and short half-life of hFGF21 resulted in low target tissue availability, which hampers its clinical application. In this study, the hFGF21 was fused with a recombinant human serum albumin (HSA), and the resulted fusion protein rHSA-hFGF21 was expressed in Pichia pastoris. After codon optimization, the recombinant gene fragment rHSA-hFGF21 was inserted into two different vectors (pPIC9k and pPICZαA) and transformed into three different strains (X33, GS115 and SMD1168), respectively. We investigated the rHSA-hFGF21 expression levels in three different strains and screened an engineered strain X33-pPIC9K-rHSA-hFGF21 with the highest expression level. To improve the production efficiency of rHSA-hFGF21, we optimized the shake flask fermentation conditions, such as the OD value, methanol concentration and induction time. After purification by hollow fiber membrane separation, Blue affinity chromatography and Q ion exchange chromatography, the purity of the rHSA-hFGF21 protein obtained was 98.18%. Compared to hFGF21, the biostabilities of rHSA-hFGF21, including their resistance to temperature and trypsinization were significantly enhanced, and its plasma half-life was extended by about 27.6 times. Moreover, the fusion protein rHSA-hFGF21 at medium and high concentration showed a better ability to promote glucose uptake after 24 h of stimulation in vitro. In vivo animal studies showed that rHSA-hFGF21 exhibited a better long-term hypoglycemic effect than hFGF21 in type 2 diabetic mice. Our results demonstrated a small-scale production of rHSA-hFGF21, which is important for large-scale production and clinical application in the future.
Animals ; Blood Glucose/metabolism* ; Diabetes Mellitus, Experimental ; Fibroblast Growth Factors ; Humans ; Hypoglycemic Agents/metabolism* ; Methanol/metabolism* ; Mice ; Pichia/metabolism* ; Recombinant Fusion Proteins ; Recombinant Proteins/metabolism* ; Saccharomycetales ; Serum Albumin/metabolism* ; Serum Albumin, Human/metabolism*