Objective : To identify the factors affecting microvascular complications among patients with type 2 diabetes (T2DM), so as to provide insights into the management of microvascular complications of T2DM. Methods: T2DM patients hospitalized in the Department of Endocrinology of a tertiary hospital in Weifang City, Shandong Province from January 2021 to January 2022 were enrolled, and subjects' basic information, lifestyle and medical history were collected using questionnaire surveys. Fasting insulin, fasting blood glucose and glycated hemoglobin were measured, and factors affecting microvascular complications were identified among T2DM patients using a multivariable logistic regression model and a decision tree model. Results: Totally 1 003 T2DM inpatients were enrolled, including 515 men (51.35%) and 488 women (48.65%), and the prevalence of microvascular complications was 40.18%. Multivariable logistic regression analysis showed that age of 60 years and older (OR=2.510, 95%CI: 1.441-4.374), T2DM duration of 10 years and longer (OR=3.205, 95%CI: 2.242-4.581), fasting insulin of lower than 3.21 μIU/mL (OR=1.749, 95%CI: 1.239-2.469), using of agents or insulin to control blood glucose (OR=1.880, 95%CI: 1.143-3.092), glycated hemoglobin level of 7% and higher (OR=1.751, 95%CI: 1.172-2.615) as factors affecting microvascular complications among T2DM patients. Decision tree analysis identified course of T2DM as a major factor affecting the risk of microvascular complications among T2DM patients, and the prevalence of microvascular complications was 70.22% among T2DM patients with disease course of 10 years and longer and fasting insulin of lower than 3.21 μIU/mL or 16.32 μIU/mL and higher, 44.23% among T2DM patients with disease course of 5 to 10 years and at ages of 60 years and older, and 43.10% among T2DM patients with disease course of less than 5 years and fasting insulin of lower than 3.21 μIU/mL. Conclusion Advanced age, long course of T2DM, low fasting insulin and high glycated hemoglobin may increase the risk of microvascular complications among T2DM patients.

Secretory anti-gp96 scFv fragment was expressed in Pichia pastoris to obtain a small molecule antibody that specifically recognizes heat shock protein gp96. The gp96-scFv fragment gene was synthesized and cloned to Pichia pastoris expression plasmid pPICZa-A. Pichia pastoris X33 was electroporated with the linearized recombinant expression vector, and expression of gp96-scFv fragment was induced by methanol. The His-tagged recombinant protein was then purified by affinity chromatography and analyzed with SDS-PAGE and Western blotting assays. The biological activities of recombinant gp96-scFv fragment were determined by Western blotting, Immunofluorescence, ELISA and FACS assays. The gp96-scFv fragment was expressed successfully in Pichia pastoris. About 50 mg of recombinant protein could be purified from 1 liter of the Pichia pastoris culture supernatant. Its molecular weight was about 15 kDa. The gp96-scFv fragment could specifically bind to gp96 protein by Western blotting, immunofluorescence, ELISA and FACS analyses. Pichia pastoris-expressed gp96-scFv fragment specifically recognizes gp96 protein, which could be used for Western blotting, Immunofluorescence, ELISA and FACS analyses.
Blotting, Western ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Membrane Glycoproteins ; immunology ; Pichia ; metabolism ; Plasmids ; Recombinant Proteins ; biosynthesis ; Single-Chain Antibodies ; biosynthesis

Objective To investigate the relationship between high-sensitivity C-reactive protein (hsCRP) and retinal vascular diameter.Methods This study was a cross-sectional study,the population was from the Kailuan study (clinical trial research registration NO.:ChiCTR-TNC-1100 1489) in 2012-2013 (age ≥ 40 years).All the people was checked by cardiovascular and epidemiological examination,of which 3 447 people had an analytical results of fundus color photography.The retinal vascular diameter was analyzed by software.Take the value of hsCRP 1％ to 99％ points and 3 363 people were finally included in the statistical analysis.The mean age of 3 363 people (male 1 880,female 1 483) was 56.69± 10.10 years old.The correlation between hsCRP and central retinal artery equivalent (CRAE),central retinal vein equivalent (CRVE),arteriovenous ratio (AVR) were evaluated by Spearman correlation coefficient.Multivariate linear regression analysis was used to futher analyze the association between hsCRP and retinal fundus diameter parameters.Results The median value of hsCRP was 1.10 (0.50,2.21) mmo/L,CRAE was (174.16±24.36) μm,CRVE was (282.70±26.32) μm,AVR was 0.62-±0.09.Spearman correlation analysis showed that hsCRP was positively correlated with CRVE (r =0.052,P =0.003) and negatively correlated with AVR (r =-0.052,P =0.002).The non-normal variables hsCRP were transformed into continuous variables lghsCRP,and CRAE,CRVE and AVR were used for multivariate linear regression analysis.After a number of factors were gradually corrected,the results showed that association between lghsCRP and CRVE (β =3.428) or AVR (β =-0.006) was statistically significant (P＜0.05).LghsCRP was a risk factor for CRVE both in men and women after the calculation of gender stratification.Conclusions CRVE increased and AVR decreased with the increase of hsCRP.The changes of retinal vascular diameter may be associated with hsCRP as a marker to predict cardiovascular and cerebrovascular disease.

Objective To analyze the clinical outcomes of a standard protocol of open reduction and internal fixation using a variable angle foot plate for a consecutive series of patients with acute Lisfranc joint injury.Methods This study retrospectively evaluated the clinical outcomes of 11 Chinese patients(13 feet) with acute Lisfranc joint injury who had been treated by open reduction and internal fixation with a variable angle foot plate from December 2016 to June 2017 at Department of Orthopaedics,West China Hospital.They were 8 men(10 feet) and 3 women(3 feet),aged from 19 to 57 years(average,36.4 years).Of them,10 were complicated with fracture of metatarsus,6 with cuneiform fracture and/or dislocation,4 with cuboid fracture,and one with cuboid fracture.According to the Myerson classification for Lisfranc fracture-dislocations,one case(one foot) belonged to type A,2(2 feet) to type Bl,5(6 feet) to type B2,2(3 feet) to type Cl and one(one foot) to type C2.The outcomes were evaluated using the visual analogue scale(VAS) and the midfoot scores of American Orthopedic Foot and Ankle Society(AOFAS).Results This cohort was followed up for 18 to 24 months(average,22 months).Their VAS scores were decreased significantly from preoperative 7.3±1.2(from 6 to 9) to postoperative 1.2±0.2(from 0 to 2)(P<0.05);their AOFAS midfoot scores were increased significantly from preoperative 0 to postoperative 84.6±4.6(P<0.05) Anatomic reduction was obtained in all the patients and all the fractures united successfully without any delayed union or nonunion.Superficial necrosis of the wound edge occurred in 2 cases without deep infection;skin anesthesia occurred in 2 and skin hypesthesia in 4,indicating a lesion of the superficial peroneal nerve.Conclusion Fixation of acute Lisfranc joint injury with a variable angle foot plate can lead to rigid stability,precise reduction and satisfactory short-term clinical outcomes.

Objective To evaluate open reduction and internal fixation (ORIF) combined with Ilizarov joint distraction in the treatment of comminuted talar body fractures.Methods Between July 2010 and May 2016,16 patients with comminuted talar body fracture underwent ORIF followed by ankle joint distraction by Ilizarov external fixator at our department.They were 9 males and 7 females,aged from 17 to 56 years (average,35.6 years).Radiological assessments were conducted by plain radiographs and computed tomography (CT) scans pre-and post-operatively.Clinical outcomes were evaluated according to visual analogue scale (VAS),American Orthopedic Foot and Ankle Society (AOFAS) hindfoot score,the short form (SF)-36 health survey scores and patients' satisfaction.Results Two patients were lost to follow-up.The remaining 14 patients were followed up for an average duration of 33.3 months (from 13 to 52 months).All the fractures united.Superficial wound infection occurred in one case.Eight patients developed post-traumatic osteoarthris;4 patients presented with avascular necrosis.At final follow-ups,the AOFAS scores ranged from 30 to 97 points (mean,69.8 points),the VAS scores from 0 to 6 points (mean,2.5 points),the physical component summary in SF-36 from 18.0 to 55 points (mean,41.7 points),and the mental component summary from 40 to 70 points (mean,54.1 points).Two patients reported strong satisfaction,5 satisfaction,4 moderate satisfaction,and 3 dissatisfaction.Conclusion For comminuted talar body fractures,ORIF combined with Ilizarov joint distraction is a new and effective treatment which can not only provide adequate fixation strength but also reduce the incidence of post-traumatic osteoarthris or avascular necrosis.

Objective To investigate in children with type 1 diabetes mellitus and provide evidence for nursing education. Methods From January to December 2016, 450 patients with T1DM were enrolled in this research,all of them were outpatients of diabetes clinic of Beijing Children′s Hospital Affiliated to Capital Medical University. The clinical data of the patients were collected through questionnaires and physical examinations to assess the insulin treatment and related factors. Results Among 450 patients, 221 were males and 229 were females, aged 9.40 (0.83-17.97) years old; body mass index was 17.43 (12.81-28.22) kg/m2; course of disease was 2.32 (0.05-12.89) years; daily insulin dosage was 0.66 (0.11-1.78) IU·kg-1·d-1; glycosylated hemoglobin (HbA1c) was 7.54 (4.4-14.9)%, of which HbA1c<7.5% accounted for 55.8%(251/450). 7.5%-9.0% accounted for 30.0(135/450)% , and >9.0% accounted for 14.2% (64/450). 44.9% ,47.1% ,8.0% (202, 212,36 cases) patients were treated respectively with routine treatment, multiple daily injection or continuous subcutaneous insulin infusion. There was no significant difference in HbA1c between patients with different insulin regimens (P>0.05), but there were significant differences in age, course of disease, body mass index and insulin dosage (P< 0.05). Conclusion Because children with type 1 diabetes have different physiological and psychological characteristics in different stages, the content of health education should be adjusted according to the patient's condition.

Objective:To evaluate the influence of manual digitorum sensory stimulation (MDSS) of hemiplegic fingers in median nerve F-wave in stroke patients.Methods:Thirty patients with hemiplegia after stroke, admitted to Department of Rehabilitation Medicine, Second Affiliated Hospital of Anhui Medical University from June 2019 to October 2022 were selected; all patients had thumb flexor modified Ashworth scale (MAS) grading≥1. Hemiplegic MDSS was given; bilateral median nerve F-wave before MDSS and median nerve F-wave at the hemiplegic side immediately after MDSS were recorded.Results:Compared with that at the healthy side before MDSS, amplitude of median nerve F-wave at the hemiplegic side was significantly increased ( P<0.05). Compared with that before MDSS, amplitude of F-wave at the hemiplegic side after MDSS in patients with thumb flexor MAS grading≤2 were significantly decreased ( P<0.05), while that in patients with thumb flexor MAS grading≥3 was significantly increased ( P<0.05). Conclusion:Amplitudes of median nerve F-wave at the hemiplegic side in patients with different thumb flexor MAS grading show that MDSS can not only inhibit the excitability of tonic motor unit related to muscle spasm, but also enhance the excitability of kinetic-motor unit mainly related to explosive power in severe muscle spasm.

ObjectiveTo investigate the effects of Buyang Huanwutang （BYWHT） on reducing inflammatory injury and improving neurological function after cerebral ischemia-reperfusion in rats via activating the adiponectin （APN） pathway. MethodMale SD rats were randomized into sham， model， BYHWT （16 g·kg^-1， twice a day）， and BYHWT + APN inhibitor （GW9662） groups. In the sham group， blood vessels were isolated. The rat model of middle cerebral artery occlusion （MCAO） was established. The rats in the BYHWT+GW9662 group was treated with subcutaneous injection of GW9662 at 4 m·kg^-1 30 min before MCAO surgery and BYHWT at 16 g·kg^-1 by gavage after MCAO surgery， once in the morning and once in the evening. The immunofluorescence （IF） assay was employed to observe the expression of adiponectin receptor 1 （AdipoR1） and the colocalization of AdipoR1 with neuronal nuclei （NeuN） and ionized calcium binding adaptor molecule 1 （Iba1） in the brain. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the expression of APN in the serum and brain. The balance beam test was carried out to examine the balance ability， and the grasping test to assess the recovery of limb strength. The immunofluorescence assay was used to detect the expression of myeloperoxidase （MPO）. Western blot was employed to determine the expression of tumor necrosis factor-alpha （TNF-α）， interleukin （IL）-1β， IL-6， and IL-10 and in the brain. ResultCompared with the sham group， the modeling promoted the expression of AdipoR1 （P<0.01）， lowered the APN levels in the serum and brain （P<0.05， P<0.01）， increased the score in the balance beam test （P<0.01）， and decreased the grasping strength of forepaw （P<0.01）， which were accompanied with increased MPO， TNF-α， IL-1β， and IL-6 levels （P<0.01） and decreased IL-10 level （P<0.01）. Compared with the model group， BYHWT promoted the expression of AdipoR1 （P<0.01）， elevated APN levels in the serum and brain （P<0.05， P<0.01）， and increased the grasping strength of forepaw （P<0.01）， which were accompanied with lowered MPO， TNF-α， IL-1β， and IL-6 levels （P<0.01） and elevated IL-10 level （P<0.01）. All the above effects were partially blocked by GW9662. ConclusionBYHWT can reduce inflammatory injury and improve neurological function in the rat model of cerebral ischemia-reperfusion by activating the APN pathway.

Objective To demonstrate the changes in flexor digitorum and extensor digitorum tension in the affected hands with shear-wave elastography (SWE) before and after manual digitorum sensory stimulation (MDSS) in hemiplegic patients with stroke. Methods A total of 51 hemiplegic post-stroke inpatients in the Department of Rehabilitation Medicine in Second Hospital of Anhui Medical University from April to June, 2020, underwent MDSS completed by a researcher who used a bare thumb and index finger to squeeze each nail bed as well as the sides of each fingertip in the affected hand. The stimulation intensity was the minimum that could cause finger extension without obvious pain, and the interval between two stimulations was five to ten seconds. Muscular tension of the flexor digitorum superficialis, flexor digitorum profundus, flexor pollicis longus and extensor digitorum were assessed with modified Ashworth Scale (MAS) and shear-wave velocity (SWV) of SWE on the affected side before and immediately after MDSS. MAS score was -1 as low muscular tension. Results The MAS scores of all the muscles significantly reduced after MDSS (|Z| > 2.843, P < 0.001); while the changes of SWV were not significantly in all the muscles with initially MAS score of 0 or -1 (t < 1.052, P > 0.05), and it reduced in those muscles with initial MAS scores of one to three (t > 2.672, P < 0.05). The SWV were positively correlated with the MAS scores both before and after MDSS (r > 0.334, P < 0.05). Conclusion MDSS can effectively, immediately, and safely relieves muscle spasms of the flexor digitorum and facilitate active finger extension in the affected hand for hemiplegic patients with stroke. SWE is useful for quantitatively and objectively evaluating muscular tension in the affected hand for hemiplegic patients with stroke.