To obtain active hFKBP52 protein for screening novel neu rotrophic drugs. Semi-nested and overlap PCR and affinity chromatography were u sed. hFKBP52 gene was cloned successfully from human fetal brain cDNA library, a nd then highly expressed (about 30%) as fusion protein in pET28a(+) vector syste m. The recombinant protein was purified as one band on SDS-PAGE. The purified h FKBP52 showed peptidyl-prolyl cis-trans isomerase (PPIase) activity, simil ar to the wild type.

Objective To investigate the bioactivities of controlled release bFGF microspheres (Ms) and their effects on the cultured osteoblasts. Methods The secondary cultured osteoblasts were divided into four groups according to the different ingredients being added to the DMEM culture medium, ie, control group,bFGF group, bFGF-PLGA-Ms group and bFGF-PELA-Ms group. The proliferation of the cultured osteoblasts was measured with cell counting method, MTT method and flow cytometry. The content of bone BGP secreted by osteoblast was also measured with RIA method. Results The in vitro cellular study showed no significant difference in the cell number and cell viability of four groups one day after plate culture.The cell number and cell viability in the bFGF-PLGA -Ms group were more than those in other three groups four and six days after plate culture. The cell number and cell viabilitythose in the bFGF group were more than those in the bFGF-PELA-Ms group six and eight days after plate culture with insignificant difference. The flow cytometrical examination showed that the G 2/M+S percentage in the bFGF group reached the highest two days after plate culture and the G 2/M+S percentage in the bFGF-PLGA-Ms group went the highest four and eight days after plate culture. Among four groups, the content of BGP in the bFGF-PLGA-Ms group was the highest and the bFGF-PELA-Ms group the next. Conclusions The effect of bFGF-PELA-Ms is not satisfactory,as indicates that the manufacturing method needs improving. However,the bFGF-PLGA-Ms can promote the proliferation and differentiation of the osteoblasts through a long period of controlled release of bFGF.

Objective: To examine the effect of early comprehensive interventions on the physical growth and nerve development among premature infants, so as to provide insights into the follow-up management of premature infants after discharge from hospital. Methods: A total of 130 premature infants delivered in Shaoxing Municipal Maternal and Child Health Care Hospital from 2019 to 2021 were selected and divided into high- and low-risk groups according to gestational age and birth weight, while 306 full-term normal infants in the same hospital during the study period served as controls. All premature infants were given early comprehensive interventions until age of 12 months, including nutritional support and parental guidance of children's feeding and development, and all normal infants received periodical health checkup according to the basic public health service program. All infants received periodical measurements of height, weight and head circumference, and the 12-month intellectual and motor development ability was measured using the Bayley Scales of Infant Development revised in Chinese cities was used to assess. Infants' physical growth, mental development index (MDI) and motor development index (PDI) were compared among groups. Results: There were 130 premature infants with gestational ages of 28 to 36 weeks and birth weight of 1 200 to 3 440 g, including 79 male infants, and there were 80 infants in the low-risk group and 50 infants in the high-risk group. The full-term infants had a gestational age of 37 to 42 weeks, and birth weights of 2 500 to 4 000 g, including 162 male infants. There were significant differences in height (Wald χ2=28.664, P<0.001) and head circumference growth (Wald χ2=19.312, P=0.013) among the three groups as revealed by the generalized estimating equation; however, no significant differences were seen in the 12-month weight (F=0.639, P=0.528), height (F=1.051, P=0.350) or head circumference (F=0.318, P=0.728) among the three groups. The percentages of abnormal MDI were 2.00%, 0 and 1.31% among the high-risk premature infants, low-risk premature infants and full-term infants at ages of 12 months (χ2=1.319, P=0.517), while the percentages of abnormal PDI were 20.00%, 7.50% and 5.56% among the three groups at ages of 12 months (χ2=12.818, P=0.002). Conclusions Following implementation of early comprehensive interventions, the premature infants have favorable physical growth and comparable MDI with full-term infants; however, a high percentage of abnormal PDI is seen in high-risk premature infants. An improvement in the motor development among high-risk premature infants is recommended to be emphasized during the management of premature infants.

Objective To explore the methods of Y-V vascular pedicle lengthening technique for cover-age of the defect of great toe after wrap-around flap transfer, and decrease the morbidity of donor site in great toes. Methods Fifteen patients received three kinds of flap for immediate resurfacing of donor defect of the great toes during wrap-around flap transfer, all flaps were harvested with Y-V vascular pedicle lengthening technique. Among them, 9 cases received the free lateral tarsal flaps transplantation for coverage of defect in donor great toes, 5 cases was repaired by medial tarsal flaps, and only one was treated by the medial plantar flap. Results All the flaps survived postoperatively. The medial plantar flap encountered the venous crisis in postoperative 3 days, and regained the normal blood supply after continuous bleeding for 2 days. All patients were satisfied with cosmetic and functional outcome in average 10- month follow-up. The appearance and sensory function of donor toe repaired by medial tarsal flaps was best among three ones. Conclusion Depending on the Y-V vascular pedicle lengthening technique, the local pedical flaps of foots are enlarged scope for coverage of great toe after wrap-around flap harvest, which can decrease the complication of donor site at the most.

Objective:To investigate the therapeutic effects of antibiotic articular cement spacer on shoulder joint infection in the elderly patients during stage-one operation.Methods:The data of 3 patients were analyzed retrospectively who had been treated at Department of Orthopaedics, The Sixth People's Hospital Affiliated to Shanghai Jiao Tong University for shoulder infection from May 2018 to December 2019. They were one man and 2 women with an average age of 65.3 years (from 64 to 67 years). One case of infection followed shoulder puncture, another proximal humeral fracture and another shoulder prosthesis replacement. All the 3 patients underwent radical debridement and implantation of antibiotic shoulder cement spacer in stage-one operation but no stage-two operation. The therapeutic effects were evaluated by American Shoulder and Elbow Surgeons (ASES) scoring, Quick Disabilities of the Arm, Shoulder, and Hand (Quick-DASH) scoring, Visual Analog Scale (VAS), and range of shoulder motion.Results:The 3 patients were followed up for 18 to 28 months (mean, 22.7 months). There was no recurrence of infection and the spacers were in good position. The microorganisms detected were Bifidobacterium brevis in one case and methicillin-resistant Staphylococcus aureus in 2 cases. At the final follow-up, the ASES scores averaged 54.4 (from 46.3 to 60.0), Quick-DASH scores 45.1 (from 40.8 to 50.0), VAS scores 2.3 (from 2 to 3), ranges of elevation 65.7 °, ranges of abduction 43.8° and ranges of external rotation 21.7°.Conclusion:For the elderly patients with shoulder infection, implantation of antibiotic shoulder cement spacer after radical debridement can well control infection, relieve pain and improve shoulder functions, sparing them secondary operation.

Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

Objective:To study the role of Jagged1 and Notch signaling pathway played in the differentiation and proliferation of RAW 264.7 cells.Methods: RAW 264.7 cells were divided into three groups to culture:The control group:RAW 264.7 cells were threated with culture and RANKL.The Jagged1 group:RAW 264.7 cells were threated with recombinant protein Jagged 1 besides the control group.The DAPT group:RAW 264.7 cells were threated with DAPT besides the Jagged 1 group.The mRNA expression of osteoclast markers(TRAP,CK,CTR) and Notch key target genes (HES-1 and HEY-1) were measured by real-time PCR.The formation of osteoclast , bone resorption , Notch expression and proliferation of RAW 264.7 cells were detected by TRAP staining , scanning electron microscope ,immunofluorescence and cell counting kit-8 ( CCK-8 ).Results: TRAP, CK, CTR , HES-1 and HEY-1 mRNA expression were significantly higher than the control group and DAPT group in Jadded 1 group ( P<0.05 ).TRAP+cell count ,osteolytic area was significantly increased in Jagged 1 group compared with control and DAPT group , and no significant difference observed between the last two groups.Immunofluorescence results showed high expression of N ICD in cell membrane and cytoplasm in all groups and additionally expressed in nucleus in Jadded 1 group.Cell proliferation was inhibited in Jagged 1 group also ( P<0.05 ).Conclusion:Jagged1 promotes RAW264.7 cells osteoclast differentiation and inhibits proliferation by activating Notch signaling pathway .

Objective To evaluate the cost-effectiveness of sequential therapy of levofloxacin in the treatment of type Ⅲa prostatitis.Methods A total of 280 in-patients diagnosed typeⅢa prostatitis were enrolled in this study and randomized to administer with either levofloxacin 500 mg( n =140 ) for intravenous injection once daily for 28 days ( control group) or administered of levofloxacin injection 500 mg ( n=140 ) once daily for 7 days followed by oral levofloxacin 500 mg once daily for 21 days( sequential group).Clinical results and safety were observed and cost-effectiveness analysis was made after 4 -week therapeutic dura-tion.Results The clinical effective rates showed no significant difference between 2 groups ( P<0.05 ).But cost -effectiveness ratio and sensitivity analysis in sequential group was significantly lower than that in control group(P<0.05).The adverse drug reaction rates in con-trol group ( 15.71%) and sequential group ( 5.71%) showed significant difference between 2 groups ( P<0.05 ).Conclusion The sequential therapy with the antibiotic therapeutic of levofloxacin 500 mg once daily achieved similar clinical efficacy to mono-therapy and offered an eco-nomic superiority and less adverse reactions in the treatment of type Ⅲa prostatitis.

Objective To compare the efficacy of Discoscope endoscope and GlideScope video laryngoscope for difficult glottis exposure.Methods Forty adult patients of both sexes scheduled for elective surgery under general anesthesia whose glottis was not visible at laryngoscopy (grade Ⅲ or Ⅳ according to Cormach-Lehane Grading of laryngoscopic view) were randomized into 2 groups (n =20 each):group GlideScope video laryngoscope (group G) and group Discoscope endoscope (group D).The glottis exposure time,intubating conditions,time from exposure of glottis to completion of tracheal intubation and incidence of postoperative sore throat and throat bleeding were recorded and compared between the 2 groups.Results Compared with group G,the glottis exposure time was significantly longer,the rate of backward pressure of cricoid cartilage lower,the time from exposure of glottis to completion of tracheal intubation shorter and the success rate of tracheal intubation at first attempt higher (P ＜ 0.05).There was no significant difference in the success rate of tracheal intubation at second attempt and postoperative incidence of sore throat and throat bleeding between the 2 groups(P ＞ 0.05).Conclusion DiscoScope endoscope is superior to GlideScope video laryngoscope in the management of difficult intubation in term of glottis exposure and success rate of tracheal intubation at first attempt.

These studies sought to evaluate the promoting effect of the exterior bFGF on membrane guided bone regeneration (MGBR). Animal models of MGBR covered with PDLLA membrane tube in bilateral radii were established in 40 New Zealand white rabbits. The membrane tubes on the left side were filled with bFGF 40 microg/100 microl and those on the contralateral side were filled with 100 microl 0.9% NaCl solution as control. The specimens were collected at 2, 4, 8, 12 weeks postoperatively. General observation, X-ray, histological grading and HE staining,and biomechanical examination were applied to studies on the repair of the models of MGBR in the two groups. Two weeks after operation, a sealed room was formed between the two bone fragments where the soft tissues covered the membrane tube. Twelve weeks after operation, PDLLA membrane became fragile and its tube shape was being maintained. Histologically, in the bFGF group numerous newly formed bone trabeculae were seen at 2 weeks after operation the radial defects had healed and the bone reconstruction and remodling had begun by the 12th week. The histological image analysis showed that the values of mean diameter and the area of new bone trabeculae in the bFGF group were higher than those in the control group (P<0.05) at 2 weeks and 4 weeks; however, there were no significant differences in these aspects between the two groups (P>0.05) at 8 and 12 weeks. The strength of the newly formed bone in the bFGF group was higher than that in the control group at 12 weeks postoperatively (P<0.05). Therefore, the authors concluded that bFGF could promote the new bone formation and biomechanical strength in the MGBR model.
Animals ; Biomechanical Phenomena ; Bone Regeneration ; drug effects ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Guided Tissue Regeneration ; Lactic Acid ; Male ; Osteogenesis ; drug effects ; Polyesters ; Polymers ; Rabbits ; Radius ; injuries ; pathology