Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

Objective:To explore the surgical procedure and effect of reconstruction of soft tissue defects in the distal segment of dorsal finger with a perforator(joint branch) flap of proper palmar digital artery combined with a dorsal metacarpal artery flap.Methods:From January 2019 to June 2022, 9 patients with soft tissue defects in distal dorsal finger, mostly with avulsion at the extensor tendon insertion point, were treated in Department of Hand and Foot Microsurgery, the Affiliated Hospital of Qingdao University. The defect areas were 1.0 cm×1.0 cm-2.5 cm×2.0 cm. Steel wires were used to reconstruct the extensor tendon insertion point. The defects were reconstructed by a perforator(joint) flap of proximal phalanx artery and a dorsal metacarpal artery flap was used to repair the donor site for proximal flap. Postoperative follow-up and evaluation included flap survival, flap appearance, scars and function of interphalangeal and metacarpophalangeal joints, through outpatient clinical, WeChat and so on.Results:The flap of digital artery perforator (joint branch) and the fasciocutaneous flap of dorsal metacarpal artery all survived. One flap of the digital artery perforator(joint branch) appeared purple and tension blisters after surgery. The stitches of the pedicle of the flap were partially dismantled at 36-48 hours after surgery. The flap was then turned to ruddy in colour 7 days after surgery. The wounds of the distal finger joint and nail bed healed well without redness and infection. The postoperative follow-up lasted for 5 to 12 months, 9 months in average. The appearance, colour and texture of the skin at distal finger were similar to those at the dorsal proximal finger and dorsal palm. Sensation recovered well, all flaps exceeding S 3. The two-point resolution (TPD) was 8-10 mm, with an average of 9.5 mm. According to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 5 cases were in excellent and 4 in good. Conclusion:The perforator(joint branch) flap of proper palmar digital artery combined with a dorsal metacarpal artery flap is simple and safe treatment. It has a fast functional recovery with an aesthetic appearance in reconstruction of the soft tissue defect in distal segment of dorsal finger.

Objective:To investigate the clinical effect of bilateral anteriolateral thigh perforator flaps (ALTPF) pedicled with descending branch of lateral circumflex femoral artery in series on reconstruction of large soft tissue defects in lower limbs.Methods:A total of 9 patients with large soft tissue defects in lower limbs were selected in the study. The patients were 6 males and 3 females aged between 18 and 57 years old. They were treated in the Department of Hand and Foot Microsurgery, the Affiliated Hospital of Qingdao University from June 2018 to January 2023. Causes of injury: 4 of traffic accident, 2 of crushing, 2 of falling from height and 1 of explosion. Five patients had combined defects in dorsal feet and 4 with tibia fractures and anterior-and-posterior-tibial soft tissue defects. All affected limbs had tissue necroses and infections in various degrees. Deep tissue defects such as tendons, nerves, bones and joints or orthopaedic implants were exposed. The areas of soft tissue defects were 30 cm×14 cm-42 cm×14 cm. All the defects were reconstructed by the bilateral ALTPF pedicled with descending branch of lateral circumflex femoral artery in series. The size of proximal flaps was 25 cm×8 cm-33 cm×13 cm and 20 cm×7 cm-29 cm×13 cm for the distal flaps. Scheduled postoperative follow-ups were conducted to observe the survival of the transferred flaps and functional recovery of calfs and feet. Sensory recovery was assessed against British Medical Research Council (BMRC) criteria. Puno scoring was used to assess the functional recovery and the effectiveness of treatment.Results:All flaps survived and the donor sites healed well. One patient developed vascular compromise within 24 hours after surgery, but it was rectified after timely surgical exploration, however it left with a small area of necrosis at distal end of the flap. The necrosis and wound healed after skin grafting in stage-two surgery. Postoperative follow-up lasted for 8.3 months in average. Two patients underwent flap thinning in stage-two surgery due to bloating appearance, 2 patients had slightly bloated flaps at the calfs, and the rest of 5 patients had satisfactory flap appearance. Two patients received nerve reconstruction by anastomosis of the lateral femoral cutaneous nerves. At the final follow-up, the sensory recovery in 9 patients achieved S 2-S 3+ according to BMRC. The average Puno score achieved 88.36 point±5.36 point, of which 4 patients in excellent, 3 in good and 2 in fair. Conclusion:The use of bilateral ALTPF pedicled with descending branch of lateral circumflex femoral artery in series to reconstruct a large soft tissue defect in lower extremity has a good clinical value, and it has become one of the effective methods.

Objective:To explore the clinical application of free chimeric medial femoral condyle osteofascial free flap (CMFCOF) in the treatment of traumatic composite bone and soft tissue defect of hand and foot.Methods:Between January, 2015 and March, 2020, 8 patients with traumatic composite bone and soft tissue defect in hand and foot were treated with CMFCOF. Of the 8 patients, there were 6 males and 2 females, with an average age of 41 (range, 24 to 56) years. The causes of injury included 3 of traffic accident, 3 of machine crush and 2 of crush. Two cases had proximal phalanx defect, 3 with metacarpal bone and 3 with metatarsal bone. The time between injury to the flap repair were 2 to 120 (mean, 84) days. The size of bone defect ranged from 2.0 cm×1.2 cm×1.2 cm to 4.4 cm× 3.0 cm×2.3 cm. The soft tissue defect ranged from 2.0 cm×1.4 cm to 5.6 cm×4.5 cm. All bone defects were on the diaphysis, without involvement of joints. Two cases had tendon defect. According to the defect of bone and soft tissue, the CMFCOF was prepared and skin graft was performed on the surface of its fascial flap.Results:The average time of flap harvesting was 53(52-96) minutes. All donor sites were directly closed. All flaps and skin grafts achieved stage I survival. All patients entered 9-16 months of follow-up, with an average of 14.5 months. The average healing time of bone was 7.5 (range, 6-10) weeks. At the last follow-up review, all flaps were not thinned. The function of donor site was restored well, without weight bearing disorder and paraesthesia in the anterior patella area. According to the trial standard of Digit Function Evaluation of the Hand Surgery Society of Chinese Medical Association, 3 patients were rated as excellent, 1 was good and 1 was fair. According to the Maryland foot evaluation criteria, 3 patients were rated as excellent for recovered with normal weight-bearing walking.Conclusion:CMFCOF can achieve satisfactory results in repairing composite bone and soft tissue defect of hand or foot. The flap has the advantages in simple operation, high quality of bone and concealed donor site.

Objective: To explore the association between dietary pattern which benefit for normal kidney function and the risk of cognitive decline or impairment in the elderly. Methods: In 2015, subjects aged 60 and over from four counties in the Nutrition and Chronic Disease Family Cohort project, were followed up in 2017. Cognitive function was repeatedly assessed, using the Mini Mental State Evaluation (MMSE) scale. Dietary pattern that benefit for normal kidney function was extracted, using the reduced rank regression method and followed by logistic regression models to explore the associations between scores that showing the kidney function on dietary patterns and the risk of cognitive deterioration and impairment in two years among those who were with normal cognition in 2015. Results: Dietary pattern that benefit for normal kidney function, was characterized by high consumption of cereal, vegetables, legume and fruits but with less meat and soy products. Comparing with the group with lowest score quartile on this dietary pattern, the risk of cognitive deterioration in the highest quartile group was significantly low (P<0.01) in two years, with an odds ratio as 0.57 (95%CI: 0.37-0.85). Linear trend was also obviously visible (P=0.007) in this group. The ones at the highest quartile group among the normal cognition ones in 2015, the risk of cognitive impairment also significantly reduced (P<0.05) in two years time, with an odds ratio as 0.52 (95%CI: 0.29-0.93). Also, linear trend could obviously be seen (P=0.01). Conclusion Dietary pattern that benefit for normal kidney function was both inversely associated with cognitive deterioration and impairment, in two years.

Objective: To assess the relationship between body mass index (BMI) and mortality in adults of Shanxi, China. Methods: Baseline data were from the '2002 China Nutrition and Health Survey’ in Shanxi province. All the death-related investigation and follow-up visits were carried out from December 2015 to March 2016. The follow-up program covered 5 360 people from all the 7 007 participants aged 18 years and over that having complete core information, with a rate as 76.5%. Participants of this study were divided into eight groups, according to the appearance of BMI. Taking the group with the lowest mortality density as the reference group, Cox regression model was used to estimate the hazard ratio (HR) and 95% confidence interval (CI) of deaths by the whole population, gender and age groups (≥60 years, <60 years). Results were then adjusted by age, gender, smoking, alcohol use and education level from the baseline survey. Sensitivity analysis was also conducted. Results: Results from the study showed that among the total number of 67 129 person- years from the average period of 12.5 years, there were 615 deaths occurred, with the mortality density as 916 per 100 000 person-years. Taking the BMI range of 26.0-27.9 kg/m² as the reference, the aHRs of death increased to 1.90 (95%CI: 1.26-2.86), 1.68 (95%CI: 1.15-2.45), 1.49 (95%CI: 1.08-2.06) and 1.72 (95%CI: 1.07-2.76) after the multivariate adjustment, in these four groups (BMI<18.5, 18.5-19.9, 22.0-23.9 and ≥30.0 kg/m²), respectively. Low body weight (BMI<18.5 kg/m²) was associated with higher risks of death in the elderly of ≥60 years, with the aHR of death as 1.94 (95%CI: 1.20-3.15). Conclusions When BMI appeared as ≤19.9 kg/m², 22.0-23.9 kg/m² and ≥30.0 kg/m², the risks of death would increase. In addition to programs that focusing on obesity, special attention should be paid to the high risk of mortality which was caused by low-weight and malnutrition in the elderly.