Aim To construct the recombinant lentiviral RNA interference(RNAi) sequence targeting rat heat shock protein 27(HSP27)gene.Methods Three complementary DNA sequences targeting rat HSP27gene were designed and synthesized.After phosphorylation and annealing,these double strands DNA were cloned into vector pSUPER-basic respectively.Next the ds-oligo DNA which expressed short hairpin RNA(shRNA) was subcloned into lentiviral vector.The transfer plasmid(pNL-HSP27-IRES2-EGFP) was constructed and confirmed by electrophoresis and sequencing.Then the three plasmids of the lentiviral vector: pNL-HSP27-IRES2-EGFP,VSVG,and pHeler were cotransfected to 293T cells by Lipofectamine 2000 to package lentiviral particles.Forty eight hours later the supernatant was collected and the titer of virus was measured by detecting expression leve1 of enhanced green fluorescent protein(EGFP).Results The analysis using restriction endonuclease digestion and sequencing confirmed that the ds-oligo DNA was successfully inserted into the lentiviral vector.The recombinant lentivirus was harvested from 293T cells with a viral titer of 3.12 ?109 fu?L-1.Vascular smooth muscle cells(VSMCs) were infected by lentivirus,and the interfering efficiency was detected by RT-PCR and Western blot.The plasmid with the highest interfering efficiency was pNL-HSP27-IRES2-EGFP-1,and the interfering efficiency was 0.771.Conclusions The recombinant lentivirus containing RNAi targeting HSP27 gene has been successfully constructed,which lay a foundation for further study of the function and gene therapy of HSP27.

In this study, the "150-cavity", next to the H5N1 influenza virus neuraminidase activity site, has been used as the target to design and synthesize a structural analogue of chlorogenic acid, N-caffeoyl-GABA, using the flexible docking simulation. The docking study showed that the N-caffeoyl-GABA could be inserted into the "150-cavity" and combined with the Arg156 side chain by hydrogen bond. The best binding free energy of H5N1 NA-N-caffeoyl-GABA complex was -7.70 kcal mol(-1), equivalent that of the NA-oseltamivir. At the same time, using the H5N1 pseudotyping virus-based NA inhibitors screening model, we determined the inhibitory effect of oseltamivir, chlorogenic acid and N-caffeoyl-GABA on the NA. Compared with chlorogenic acid, N-caffeoyl-GABA significantly enhanced the inhibitory effect on NA, but less than oseltamivir. This study showed that the "150-cavity" could possibly be used as a new neuraminidase inhibitors target, and provided a path for the development of new neuraminidase inhibitors.

Objective To explore the effect of functional electrical stimulation combined with treadmill training and botulinum toxin type A injection on foot-drop and strephenopodia among stroke survivors.Methods Sixty-seven stroke survivors with foot-drop and strephenopodia were randomly divided into an electrical stimulation group (n=23),a conventional treatment group (n =22) and a combined treatment group (n =22).All 3 groups received a 400 U injection of BTX-A and electrical stimulation.After 24 hours,the patients in the conventional treatment group received conventional treatment including a brain protection agent,limb function exercises,gait training,balance training and training in the activities of daily living.The patients in the combined treatment group received that conventional treatment,plus functional electrical stimulation and weight loss training on a treadmill.The patients in the electrical stimulation group received functional electrical stimulation treatment supplementing the conventional treatment.Therapeutic effects were evaluated before and after six weeks of treatment using integral electromyography (iEMS) of the anterior tibial muscle and the lateral head of the gastrocnemius muscle,the co-contraction ratio (CR) during ankle dorsiflexion,the modified Ashworth Scale (MAS),the Berg balance scale (BBS),a functional walking score (FAC),and the active range of motion (AROM) of the ankle in dorsiflexion and eversion.Results After the treatment,significant improvement was observed in all three groups in the average iEMS value of the anterior tibial muscle and the lateral head of the gastrocnemius muscle,the CR in ankle dorsiflexion,and in their MAS,BBS,FAC and AROM results.There was no significant difference among the three groups after treatment in their average iEMS values at the lateral head of the gastrocnemius.The average values of the other indicators were,however,significantly better in the combined treatment group than in the other 2 groups.Conclusion Functional electrical stimulation combined with treadmill training and botulinum toxin type A injection can significantly improve foot-drop,strephenopodia and the walking function of stroke survivors.This combined treatment deserves popularization and application in clinical practice.

Objective To study the clinical characteristics and treatment of esophageal injury caused by button battery foreign bodies in children, then improve the diagnosis and management level of this hazardous problem of children. Methods 15 patients with esophageal injury caused by button battery foreign bodies were retrospectively analyzed the clinical characteristics, treatment and prognosis. Results 12 button battery foreign bodies were removed successfully with rigid esophagoscope, 3 with electronic gastroscope. 12 cases occurred serious esophageal lesions, 10 patients were fully recovered after nasal feeding, anti-inflammatory and supporting therapy. Serious complication such as esophageal perforation occurred in 2 cases, one of them occurred tracheo-esophageal fistula. Conclusion Button battery embedded in the esophagus is easy to cause serious esophagus injury, early detection is the key of the management, and its prompt removal is mandatory. Button battery have a distinctive appearance on radiography, chest radiographs can be used as the first choice of diagnosis. The reasonable treatment can obtain better curative results and avoid serious complications according to the course of the disease and esophageal damage degree. Physicians must recognize the hazardous potential and serious implications such as esophageal perforation and tracheo-esophageal fistula.

Objective To investigate the preventive use of antibiotics for internal fixation as selective removal surgery during perioperative period.Methods The sex, age, routine blood examination data, preventive medications, secondary incision infection and postoperative hospital stay for the patients underwent selective removal surgery for internal fixation from July2015to June 2017were analyzed.Results The average postoperative hospitalization time for the preventive medication group (n=71) and the untreated group (n=350) were (1.80±1.24) days and (1.27±0.61) days.This difference was statistically significant (P＜0.05).Conclusion Preventive antibiotics use is not related to incision infections.It is recommended to correct the current behavior of the preventive drug use.The preventive antibiotics use in selective removal surgery for internal fixation in the Department of orthopedics needs to be reduced.

Objective: To summarize and explore the clinical effect and application of L-shaped flap pedicled with peroneal artery perforator in the treatment of unhealed lateral L-shaped incision after calcaneal fractures. Methods: From October 2013 to March 2015, 17 patients with unhealed L-shaped incision after calcaneal fractures were treated with one-stage thorough debridement, artificial bone filling with antibiotics-laden calcium sulphate and L-shaped flap pedicled with peroneal artery. Flap areas ranged from 8 cm × 2 cm to 11 cm × 3 cm. Donor sites are primarily sutured. Results: All 17 patients were followed up for 6 to 18 months with an average of 11.5 months. All patients with infection were cured effectively, demonstrating well survived flaps with normal elasticity and pigmentation, no scar contracture, satisfactory appearance and normal ankle joint motility. Conclusions L-shaped flap pedicled with peroneal artery perforator was an effective method in the treatment of unhealed lateral L-shaped incision after calcaneal fractures. It was suitable for coverage of the unhealed wound and worthy of being popularized.

OBJECTIVETo investigate the relationship between the mean CT value and bone density value and explore a rapid and accurate image segmentation method for lumbar CT values in different age groups.

METHODSWe randomly collected thin lumbar spine CT scan data from 202 patients stratified by gender and age (at the interval of 10 years). The data of each group were imported into Mimics 14.0 software to obtain independent Mask files using the new image segmentation method. The Mask files were used to fill the empty space and calculate mean, maximum, and minimum CT values.

RESULTSIn the age groups of 21-30, 31-40, 41-50, 51-60, and 61-70 years, the male subjects had mean CT values of 424.94 ± 52.23 Hu, 405.59 ± 53.60 Hu, 361.76 ± 53.50 Hu, 344.17 ± 47.76 Hu, and 332.88 ± 58.33 Hu, respectively; the values in the female group were 439.89 ± 50.99 Hu, 448.06 ± 65.89 Hu, 421.43 ± 54.74 Hu, 369.07 ± 78.27 Hu, and 304.98 ± 55.05 Hu, respectively. The mean CT value in men aged 61-70 years was significantly lower than the values in men below 50 years (P<0.05), but comparable with that in those aged 51-60 years (P>0.05); The mean CT value was significantly lower in women aged 51-70 years groups than in those aged below 50 years (P<0.05).

CONCLUSIONSThe mean CT values are correlated with the bone density values. The new image segmentation method allows rapid and accurate acquisition of the CT values in the regions of interest. The lumbar mean CT values tend to decrease with age.

Adult ; Aged ; Aged, 80 and over ; Bone Density ; Female ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; Male ; Middle Aged ; Software ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo investigate the application of 3D printing and digital technology in preoperative assessment and planning of internal fixation surgery for complex tibial plateau fracture.

METHODSComplex tibial plateau fractures and commonly used plates for tibial plateau were imaged using computed tomography (CT) to reconstruct the 3D fracture and plate models. The 3D models were used to perform virtual reduction and preoperative planning of internal fixation surgery with the most appropriate plates assisted by the 3D library of plates. According to the optimal plan, the 3D physical models of tibial plateau fractures and plates were 3D printed to simulate internal fixation operation. The effects of internal fixation were compared between the virtual surgery and the simulated surgery based on the 3D models.

RESULTSThe effects of internal fixation in the simulated surgery based on the 3D models were consistent with those of the virtual surgery. No significant difference was found in the screw length between the two surgeries.

CONCLUSIONThe combination of 3D printing and digital design can improve the effects of internal fixation for complex tibial plateau fractures.

Bone Plates ; Bone Screws ; Fracture Fixation, Internal ; Fractures, Bone ; Humans ; Printing, Three-Dimensional ; Tibial Fractures ; surgery ; Tomography, X-Ray Computed

Objective:To explore the value of transesophageal echocardiography(TEE) guidance for transcatheter DragonFly? system edge-to-edge tricuspid regurgitation (TR) repair.Methods:Five cases who were chosen in the Second Affiliated Hospital, Zhejiang University School of Medicine from December 2020 to January 2021 with surgical high-risk and severe functional TR underwent transcatheter DragonFly edge-to-edge repair with the guidance of TEE. Preoperative TEE was used to evaluate the tricuspid valve anatomy and the origin and etiology of regurgitation in detail; intra-procedure guidance of TEE was performed during the DragonFly system for tricuspid valve edge-to-edge repair intervention and after release of the DragonFly clip, the effect of surgery was assessed immediately and compared with pre-procedure TEE.Results:A total of 10 DragonFly clips were implanted in 5 patients (3 in each of patients, 2 in 1 patient, and 1 in each of patients). One of the 3 clips in 1 patient fell off unilaterally from the septal valve after release, and the other 9 clips were well positioned and fixed. Immediately post-operation assessment by TEE depicted the TR in 3 patients declined to mild and 2 to moderate. The vena contracta area by using three-dimensional color blood flow quantitative assessment was reduced[(0.93±0.26)cm 2 vs (0.20±0.11)cm 2]. No complications such as serious tricuspid valve injury, pericardial tamponade, thromboembolism occurred in the 5 patients. Conclusions:TEE plays an important role in guiding and monitoring transcatheter DragonFly system edge-to-edge TR repair during the entire procedure.

Objective To study the expression of epidermal growth factor receptor 1 (EGFR) protein in breast cancer and its correlation to molecular subtyping and hormone receptor status.Methods 467 cases of breast cancer were included.According to ER,PR,HER2,and Ki-67 status,the cases were categorized into 4 molecular subtypes,including 185 cases of luminal A,109 cases of luminal B,76 cases of HER2-enriched,and 70 cases of triple-negative breast cancer (TNBC).According to ER and PR status,the cases were divided into 4 subtypes,including 240 cases of ER+/PR+,50 cases of ER+/PR-,4 cases of ER-/PR+,and 173 cases of ER-/PR-.Results EGFR protein expression rates in Luminal A,Luminal B,HER2-enriched and TNBC were 16.8％(31/185),54.1％(59/109),97.4％(74/76),78.6％(55/70),respectively.The EGFR expression in HER2-enriched was significantly higher than those in TNBC,Luminal B and Luminal A(P＜0.01),and EGFR expression in TNBC was significantly higher than those in Luminal B and Luminal A (P＜0.01),furthermore,EGFR expression in Luminal B was significantly higher than that in Luminal A (P＜0.01).EGFR protein expression rates in ER+/PR+ subtype,ER+/PR-subtype,ER-/PR+ subtype and ER-/PR-subtype were 25.4％ (61/240),52.0％ (26/50),75.0％ (3/4),88.4％(153/173),respectively.The EGFR expression in ER-/PR-subtype was significantly higher than in ER+/PR+ subtype and ER+/PR-subtype (P＜0.01),and EGFR expression in ER+/PR-subtype was significantly higher than that in ER+/PR+ subtype (P＜0.01).EGFR protein expression rate was higher in ER-/PR-subtype than in ER-/PR+ subtype,and EGFR protein expression rate was higher in ER-/PR+ subtype than that in ER+/PR+ subtype and ER+/PR-subtype,but all of the difference were not statistically significant (P＞0.05).Conclusion EGFR protein expression is closely related to breast cancer molecular subtyping and negative hormone receptor expression,which is a potential biomarker of anti-breast cancer therapy.