Transcriptomic and physiological analyses of Symphytum officinale L. in response to multiple heavy metal stress.

Soil heavy metal contamination has become a global environmental issue, which threaten soil quality, food security and human health. Symphytum officinale L. have exhibited high tolerance and restoration capacity to heavy metals (HMs) stress. However, little is known about the mechanisms of HMs in S. officinale. In this study, transcriptomic and physiological changes of S. officinale response to different HMs (Pb, Cd and Zn) were analyzed and investigated the key genes and pathways involved in HMs uptake patterns. The results showed that phenotypic effects are not significant, and antioxidant enzyme activities were all upregulated. Transcriptome analysis indicated that 1247 differential genes were up-regulated, and 1963 differential genes were down-regulated under Cd stress, while 3752 differential genes were up-regulated, and 7197 differential genes were down-regulated under Pb stress; and 527 differential genes were up-regulated; and 722 differential genes were down-regulated under Zn stress. Based on their expression, we preliminarily speculate that different HMs resistance of S. officinale may be regulated by the differential expression of key genes. These results provide a theoretical basis for determining the exact expression of genes in plants under different heavy metal stress, the processes involved molecular pathways, and how they can be efficiently utilized to improve plant tolerance to toxic metals and improve phytoremediation efficiency.

Exploring the catalytic diversity of two short-chain dehydrogenases/reductases from Stachybotrys chartarum.

Short-chain dehydrogenase/reductases (SDRs) belong to the NAD(P)(H)-dependent oxidoreductase superfamily, which have various functions of catalyzing oxidation/reduction reactions and have been generally used as powerful biocatalysts in the production of pharmaceuticals. In this study, ScSDR1 and ScSDR2, two new SDRs have been identified and characterized from Stachybotrys chartarum 3.5365. Substrate scope investigation revealed that both of the enzymes possessed the ability to oxidize ß-OH to ketone specifically, and exhibited substrate promiscuity and high stereo-selectivity for efficiently catalyzing the structurally different prochiral ketones to chiral alcohols. These findings not only suggest that ScSDR1 and ScSDR2 might be potent synthetic tools in drug research and development, but also provide good examples for further engineered enzymes with higher efficiency and stereo-selectivity.

A new combination and a new synonym of Gesneriaceae in China.

Didymocarpussubpalmatinervis W.T.Wang was shown to be conspecific with Petrocodonlithophilus Y.M.Shui, W.H.Chen & Mich.Möller, by checking original literature, examining specimens, tracing specimen collecting history, and conducting field surveys. The results show morphological characteristics and geographical distribution overlaps between these two species. The transfer of Didymocarpussubpalmatinervis to Petrocodon as a new combination with Petrocodonsubpalmatinervis (W.T.Wang) F.Wen & Z.L.Li is proposed here, and P.lithophilus is synonymized with P.subpalmatinervis. Color photographs and essential information are also provided, including a detailed comparison of description, distribution, habitat, and the proposed IUCN conservation status.

[Clinical Analysis of 17 Adult Patients With Fulminant Type 1 Diabetes Mellitus].

Objective: To explore the clinical characteristics of adult patients with fulminant type 1 diabetes mellitus (FT1DM), a specific subtype of type 1 diabetes mellitus (T1DM). Methods: We collected the clinical data of patients who were admitted to West China Hospital, Sichuan University in 2010-2019 for FT1DM and type 1 diabetes mellitus (T1DM) presenting with diabetic ketoacidosis (DKA) at the onset. In addition, all the FT1DM patients were followed up. Results: A total of 70 patients presenting with DKA at the onset of T1DM were admitted to and received treatment at West China Hospital in 2010-2019. Among them, 17 (24.3%) had FT1DM and 53 did not. The mean ages of the FT1DM patients and the non-FT1DM patients were (33.2±12.8) years and (27.5±11.2) years, and the mean body mass indices were (22.6±2.9) kg/m 2 and (19.2±2.9) kg/m 2, respectively. A total of 14 FT1DM cases had symptoms of upper respiratory tract infection or acute gastroenteritis before the onset of the disease and 4 cases were related to pregnancy. The median time from the onset of the disease to the first diagnosis of DKA of the FT1DM group (median [P 25-P 75]: 2 [1-4] days, P<0.001) was significantly shorter than that of the non-FT1DM group (median [P 25-P 75]: 30 [17-78] days). The mean maximum blood glucose levels at the time of the first visit to the doctor of the FT1DM patients ([39.9±11.4] mmol/L, P<0.001) were significantly higher than that of the non-FT1DM patients ([28.9±9.2] mmol/L), but the HbA1c (6.6%±0.6%, P<0.001) and glycosylated serum albumin (GA) (21.4%±3.0%, P=0.001) levels of the FT1DM patients were significantly lower than those of the non-FT1DM group (HbA1c: 12.8%±2.7%; GA: 44.8%±15.0%). The median serum amylase in the FT1DM group was significantly higher than that in the non-FT1DM group (101 [54-336] IU/L vs. 54 [42-166] IU/L, P=0.045) and the median serum lipase in the FT1DM group showed a trend of being higher than that in the T1DM group (81 [57-154] IU/L vs. 46 [28-195] IU/L, P=0.051). 8.7% of the non-FT1DM patients tested positive for anti-glutamic acid decarboxylase antibody (GAD-Ab), while the FT1DM patients all tested negative. At the time of discharge, the mean daily insulin dose of the FT1DM patients was (0.67±0.22) IU/kg, which was not significantly different from that of the non-FT1DM group ([0.74±0.29] IU/kg, P=0.349). After about 6.5 years of follow-up, the mean daily insulin dose of the FT1DM patients was (0.73±0.19) IU/kg, which was similar to the insulin dosage on discharge ( P=0.409). Conclusion: Compared with the non-FT1DM patients presenting with DKA at the onset, FT1DM patients have fewer typical diabetic symptoms, lower fasting C-peptide levels, higher serum amylase levels, and increased incidence of vomiting or other symptoms of gastrointestinal infections, and are more likely to be misdiagnosed. Therefore, it is very important for clinicians to correctly identify FT1DM as early as possible and administer early and long-term insulin replacement therapy.

Mechanism and intervention of murine transfusion-related acute lung injury caused by anti-CD36 antibodies.

Anti-CD36 Abs have been suggested to induce transfusion-related acute lung injury (TRALI) upon blood transfusion, particularly in Asian populations. However, little is known about the pathological mechanism of anti-CD36 Ab-mediated TRALI, and potential therapies have not yet been identified. Here, we developed a murine model of anti-CD36 Ab-mediated TRALI to address these questions. Administration of mouse mAb against CD36 (mAb GZ1) or human anti-CD36 IgG, but not GZ1 F(ab')2 fragments, induced severe TRALI in Cd36+/+ male mice. Predepletion of recipient monocytes or complement, but not neutrophils or platelets, prevented the development of murine TRALI. Moreover, plasma C5a levels after TRALI induction by anti-CD36 Abs increased more than 3-fold, implying a critical role of complement C5 activation in the mechanism of Fc-dependent anti-CD36-mediated TRALI. Administration of GZ1 F(ab')2, antioxidant (N-acetyl cysteine, NAC), or C5 blocker (mAb BB5.1) before TRALI induction completely protected mice from anti-CD36-mediated TRALI. Although no significant amelioration in TRALI was observed when mice were injected with GZ1 F(ab')2 after TRALI induction, significant improvement was achieved when mice were treated postinduction with NAC or anti-C5. Importantly, anti-C5 treatment completely rescued mice from TRALI, suggesting the potential role of existing anti-C5 drugs in the treatment of patients with TRALI caused by anti-CD36.

[Clinical Characteristics and Prognosis of Diabetic Foot Ulcers Patients of Different Renal Function Statuses].

Objective: To explore the clinical characteristics and the prognosis of diabetic foot ulcers (DFU) inpatients of different renal function statuses. Methods: A retrospective analysis of 962 inpatients with DFU was conducted. The patients were divided into three groups according to their renal function statuses, and the clinical characteristics of the three groups were compared to identify differences. In addition, the patients were followed up in outpatient clinics or by telephone and their prognostic status and risk factors for death were analyzed. Results: Analysis of the clinical characteristics showed that, compared with diabetic patients with normal renal function or mild renal function impairment, diabetic patients with moderate and severe renal function impairment had a longer course of disease ( P<0.001). Patients with foot ulcers of Wagner grade 4 predominates the moderate and severe renal function impairment groups ( P<0.05). Patients in the moderate and severe renal function impairment groups had a relatively higher proportion of comorbidities, including hypertension, coronary heart disease, and peripheral arterial disease ( P<0.05). These patients had relatively lower levels of glycosylated hemoglobin and hemoglobin (all P<0.05) and relatively higher levels of neutrophil ratio and procalcitonin (all P<0.05). Of the two groups, patients in the moderate renal function impairment group were older ( P<0.001) and had lower ankle-brachial index ( P<0.001). The severe renal function impairment group had a higher proportion of patients with foot ulcers of Wagner grades 3 and 5 (all P<0.05). For the purpose of conducting prognostic analysis, 748 patients were followed up in outpatient clinics or by telephone for a median length of 41 months. Among them, 239 died. The all-cause mortality was 31.9%, and the mortality in the three groups was 25.8%, 46.2% ( P<0.001), and 59.4% ( P<0.001), respectively. The survival rate of patients in the moderate and severe renal function impairment groups was significantly lower than those in the normal renal function and mild renal function impairment groups ( P<0.001). Univariate Cox regression analysis showed that age, concomitant coronary heart disease and peripheral arterial disease, degree of renal function impairment, and foot ulcers of Wagner grade 4 and 5 were associated with all-cause deaths. Furthermore, multivariate Cox regression analysis showed that moderate and severe renal function impairment was an independent risk factor for all-cause deaths in DFU patients ( P<0.001). Conclusions: As renal function impairment worsens, patients with DFU present clinical characteristics of greater complexity, higher risks of cardiovascular events, and higher mortality. It is essential to prevent kidney damage and foot ulcers, to pay attention to the cardiovascular risks of DFU patients with moderate and severe renal function impairment, and to reduce mortality.

[Preliminary Application of Contrast-Enhanced Ultrasound in the Treatment of Diabetic Ulcers].

Objective: To investigate the role of contrast-enhanced ultrasound (CEUS) in the treatment of diabetic ulcers. Methods: The clinical data of 27 diabetic patients, who underwent CEUS examination of their ulcers in our hospital between April 2021 and July 2022 were collected. Among them, 26 patients suffered from diabetic foot ulcers, 5 of whom underwent amputation during hospitalization, and one patient suffered from hip ulcer. The 27 patients' mean age was (64.08±12.57) years. Fasting blood glucose levels of the patients were 3.36-34.61 mmol/L, with a mean of (10.62±8.77) mmol/L. Their glycosylated hemoglobin levels were 5.80%-10.70%, with an average of 7.96%±1.50%. Philips EPIQ7 ultrasound system with L9-3 linear probe of 3-9 MHz was used. First, the patients' ulcers were examined with conventional ultrasound to observe for abnormal echo. Then, 2.4 mL SonoVue (Bracco, Italy), a contrast agent, was injected intravenously through the elbow to look for effusion/pus, sinus tract, or dead space in the lesion area, and images were acquired. Results: Among the 27 patients, except for 5 with amputation stumps, 22 patients had wound areas ranging from 0.16 cm 2 to 215 cm 2, all being accompanied by sinus tract formation. Ten patients underwent ultrasound examination during their treatment. The positive rate of the results of conventional ultrasound was 50% (5/10) for identifying effusion/pus and pseudoaneurysm in the deep area of ulcers, while the positive rate of CEUS results was 100% (10/10). In addition to the lesions found by conventional ultrasound, CEUS also found large sinus tracts or dead spaces in the deep surface of ulcers in 5 additional patients. Of the 27 patients, 17 underwent ultrasound examination of the healing status of sinus tracts and dead spaces in the deep areas of ulcers before discharge. No sinus tracts in the deep areas of the ulcers were found by conventional ultrasound. However, relatively small dead spaces or sinus tracts in the deep areas of the ulcers were found in 10 patients by CEUS. Conventional ultrasound and CEUS found that 1 patient had a small amount of fluid in the amputation stump. In the remaining 6 patients, no deep sinus tracts in the ulcers were found by either conventional ultrasound or CUES, and the ulcers healed completely. Conclusion: By examining microvascular perfusion in diabetic wounds with CEUS, we can observe the extent of sinus tracts during treatment and whether the sinus tracts have healed or whether there are still dead spaces before patient discharge, which provides support for clinical decision-making concerning the treatment of diabetic ulcers.

Enzymatic synthesis of anhydroicaritin, baohuoside and icariin.

Anhydroicaritin (1a), baohuoside (1b) and icariin (1c) were recognized as major pharmacologically active ingredients of Epimedium plants. Their primary means of acquisition were chemical isolation from plants. However, it suffers from low yield, environmental pollution and shortage of plants. Herein, to remedy these problems, biosynthesis was explored to obtain the three active ingredients. Fortunately, with SfFPT as 8-prenyltransferase, EpPF3RT and Ep7GT as glycosyltransferases, kaempferide (1) was transferred to 1a, 1b and 1c enzymatically. Thus, we report the details of this method. This approach represents a promising environmental friendly alternative for the production of these compounds from an abundant analogue.

[Clinical Characteristics of Diabetic Patients with Initial and Recurrent Foot Ulcers].

Objective: To explore the risk factors for the recurrence of foot ulcers by analyzing clinical characteristics of the patients with diabetic foot ulcers (DFU) in West China Hospital, Sichuan University. Methods: A retrospective analysis was carried out with the clinical data of 817 DFU patients hospitalized at West China Hospital, Sichuan University between January 1, 2012 and December 31, 2020. The patients were divided into an initial ulceration group (502 cases) and a recurrent ulceration group (315 cases) according to their history of foot ulcers. The differences in clinical characteristics between the two groups were compared, and multivariate logistic regression analysis was conducted to identify the risk factors associated with the recurrence of foot ulcers. Results: Initial and recurrent DFU patients both had predominantly neuro-ischemic foot ulcers, and the most common sites of ulceration were the first and fifth toes in both groups. Compared with the initial DFU group, more patients in the recurrent group had foot ulcers of Wagner grade 3 and ulcerous wounds located on calluses ( P<0.05), and fewer patients in the recurrent group suffered from foot gangrene ( P<0.05). Patients with recurrent DFU had lower glycated hemoglobin, platelet counts, and fibrinogen levels ( P<0.05), and higher serum uric acid and creatinine levels ( P<0.05). Hemoglobin, white blood cell count, estimated glomerular filtration rate, erythrocyte sedimentation rate and C-reactive protein levels were not significantly different between the two groups ( P>0.05). Multivariable logistic regression analysis showed that male sex ( OR=1.555, 95% CI: 1.097-2.204, P=0.013), duration of diabetes≥10 years ( OR=2.369, 95% CI: 1.473-3.810, P<0.001), history of amputation ( OR=4.518, 95% CI: 2.386-8.553, P<0.001), foot osteoporosis ( OR=1.711, 95% CI: 1.065 to 2.751, P=0.027), ulcerous wound located on foot callus ( OR=1.786, 95% CI: 1.058-3.012, P=0.030), and coronary heart disease ( OR=0.668, 95% CI:0.453-0.987, P=0.043) were significantly associated with the recurrence of foot ulcers. Conclusions: Male sex, duration of diabetes being over 10 years, history of previous amputation, foot osteoporosis, and ulcerous wounds located on foot callus are independent risk factors of recurrent foot ulcers in patients with DFU. Therefore, even after their foot ulcers have healed, special attention should be given to the care of foot for patients with DFU, which may reduce the recurrence of foot ulcers.

[Single-Center Experience of Treating Chronic Limb-Threatening Ischemia of Lower Limbs Combined with Diabetes].

Objective: To summarize our hospital's single-center experience of and reflections on the treatment of chronic limb-threatening ischemia (CLTI) of lower limbs combined with diabetes in the past 5 years. Methods: We retrospectively analyzed cases of lower limb CLTI combined with diabetes diagnosed at our hospital from March 2017 to June 2021. The baseline data, surgical information, and follow-up results of the patients were collected. The primary outcome indicator was the patency rate of lower limb target artery within 1 year post-op, and the secondary indicators were the reoperation rate within 1 year post-op and the amputation rate within 1 year post-op. Results: A total of 89 patients with lower limb CLTI combined with diabetes were included in the study. A total of 85 patients underwent percutaneous transluminal angioplasty and the operation of 7 patients ended in failure, with the operation success rate reaching 91.76% (78/85). Three patients underwent femoral popliteal artery bypass grafting with artificial blood vessels and one patient underwent iliac femoral artery bypass grafting with artificial blood vessels, with the success rate of the operations reachign 100% (4/4). Among 78 patients who successfully underwent percutaneous transluminal angioplasty, the median follow-up time was 33 months (13, 64). Two patients died within one year after operation, with the post-op one-year survival rate being 97.44% (76/78). The post-op 1-year reoperation rate was 19.23% (15/78), the 1-year target vascular patency rate (deaths not included) was 85.53% (65/76), and the 1-year amputation rate was 3.85% (3/78). Among the patients who underwent bypass surgery, the follow-up period was 13-48 months. No thrombosis in or re-occlusion of the artificial blood vessels were observed during the follow-up period, and the artificial blood vessels remained unoccluded. Conclusion: Transluminal angioplasty has a relatively ideal rate of postoperative vascular patency. In addition, it is a minimally invasive procedure involving low perioperative risks and is performed under local anesthesia. Therefore, it can be used as the preferred treatment for patients with CLTI. On the other hand, bypass surgery has good long-term patency rate, but it involves higher perioperative risks and the procedure is more invasive. Therefore, bypass surgery can be used as an alternative when transluminal angioplasty ends in failure.

Protective effects of fowl-origin cadmium-tolerant lactobacillus against sub-chronic cadmium-induced toxicity in chickens.

Cadmium (Cd) directly endangers poultry health and indirectly causes harm to human health by food chain. Numerous studies have focused on removing Cd using lactic acid bacteria (LAB). However, there is still a lack of in vivo studies to validate whether Cd can be absorbed successfully by LAB to alleviate Cd toxicity. Here, we aimed to isolated and screened poultry-derived Cd-tolerant LAB with the strongest adsorption capacity in vitro and investigate the protective effect of which on sub-chronic Cd toxicity in chickens. First, nine Cd-tolerant LAB strains were selected preliminarily by isolating, screening, and identifying from poultry farms. Next, four strains with the strongest adsorption capacity were used to explore the influence of different physical and chemical factors on the ability of LAB to adsorb Cd as well as its probiotic properties in terms of acid tolerance, bile salt tolerance, drug resistance, and antibacterial effects. Resultantly, the CLF9-1 strain with the best comprehensive ability was selected for further animal protection test. The Cd-tolerant LAB treatment promoted the growth performance of chickens and reduced the Cd-elevated liver and kidney coefficients. Moreover, Cd-induced liver, kidney, and duodenum injuries were alleviated significantly by high-dose LAB treatment. Furthermore, LAB treatment also increased the elimination of Cd in feces and markedly reduced the Cd buildup in the liver and kidney. In summary, these findings determine that screened Cd-tolerant LAB strain exerts a protective effect on chickens against sub-chronic cadmium poisoning, thus providing an essential guideline for the public health and safety of livestock and poultry.

Analysis of endophyte diversity of two Gentiana plants species and the association with secondary metabolite.

BACKGROUND: The influence of external environmental factors on secondary metabolites of medicinal plants has always been studied. However, little is known about the relationships between endophytes and host metabolites, especially the relationship differences between different plant species. Thus, we used high-throughput sequencing methods to compare endophyte diversity from roots of two closely related species, Gentiana officinalis and G. siphonantha, from the same production area, and analyze the association with four secondary metabolites (Gentiopicroside, Loganic acid, Swertiamarine and Sweroside). RESULTS: The fungal and bacteria communities' richness and diversity of G. siphonantha was higher than G. officinalis. Ascomycota and Proteobacteria were dominant fungal and bacterial phylum of the two closely related species. At the genus level, Tetracladium and Cadophora were dominant fungal genus in G. officinalis and G. siphonantha samples, respectively. While Pseudomonas was dominant bacterial genus in two closely related species, with relative abundances were 8.29 and 8.05%, respectively. Spearman analysis showed that the content of loganic acid was significantly positively correlated with endophytic fungi, the content of gentiopicroside, swertiamarine and sweroside were significantly positively correlated with endophytic bacteria in the two related species. PICRUSt and FUNGuild predictive analysis indicated that metabolism and saprotroph was primary function of endophytic bacteria and fungi in the two related species. CONCLUSION: Our results will expand the knowledge on relationships of plant-microbe interactions and offer pivotal information to reveal the role of endophytes in the production of Gentiana plant and its important secondary metabolite.

Two new 5/6/6 polyketide-amino acid hybrids from a genetic mutant of Periconia sp. F-31.

Peridecalins C and D (1 and 2), one decalin and one oxygen-decalin containing polyketide-amino acid hybrids with 5/6/6 ring system, was isolated from a genetic mutant of Periconia sp. F-31. Their structures were elucidated through extensive spectroscopic data analysis, including 1 D/2D NMR and HR-MS spectra. Biosynthetically, two proposed Diels-Alder reactions are supposed to be involved in the skeleton construction of 1 and 2.

Dynamic observation on collateral circulation construction of patient with vertebral artery restenosis after stenting: case report.

Purpose/aim of the study: Posterior circulation stroke (PCS) accounts for 20% of ischemic stroke, and vertebrobasilar stenosis is an important cause of PCS. Notably, not all patients with artery stenosis progress to ischemic stroke, and one of the important reason is that collateral circulation construction plays important protection role in this process.Clinical presentation: Here, we present the case of a 71-year-old male who presented with lightheadedness and three episodes of loss of consciousness after bilateral subclavian artery stenting. Digital subtraction angiography (DSA) demonstrated severe stenosis of the left vertebral artery, and the bilateral subclavian artery was kept open. The patient was then given the left vertebral artery stenting in an effort to resolve the vascular stenosis. As expected, he achieved a complete remission after stenting. However, 6 months later the patient suffered from loss of consciousness again. Repeat DSA confirmed restenosis of the left vertebral artery, and revealed a collateral flow to the left vertebral artery which fed by external carotid collateral branches. Then DSA was performed after 12 months, and another collateral circulation involving thyrocervical trunk was also found supplying flow to the left vertebral artery. In this process, the frequency of loss of consciousness gradually decreased as the collateral circulation construction. Conclusion: Through this case, we observe the whole process of the collateral circulation construction. Moreover, this case serves as a testament to the variability and complexity of vertebrobasilar arteriopathies, suggesting promotion of collateral flow offers the opportunity for outcome improvement.

Corolla Abscission Triggered by Nectar Robbers Positively Affects Reproduction by Enhancing Self-Pollination in Symphytum officinale (Boraginaceae).

Nectar robbers, which affect plant fitness (directly or indirectly) in different degrees and in different ways, potentially constitute a significant part of mutualistic relationships. While the negative effects of nectar robbing on plant reproductive success have been widely reported, the positive effects remain unknown. The target of our study was to evaluate the effects of nectar robbers on the reproductive success of Symphytum officinale (Boraginaceae). We observed the behavior, species and times of visitors in the field, and we assessed the effect of nectar robbers on corolla abscission rate and time. To test the fitness of corolla abscission, we detected the changes in stigma receptivity, pollen viability, pollen amount and appendage opening size along with the time of flower blossom. The flowering dynamics and floral structure were observed to reveal the mechanism of self-pollination. Finally, pollen deposition seed set rate and fruit set rate were determined to estimate the effect of nectar robbers on reproduction success. We observed 14 species of visitors and 2539 visits in 50 h of observation; 91.7% of them were nectar robbers. The pressure and nectar removal of nectar robbers significantly promoted corolla abscission during a period when pollen grains are viable and the stigma is receptive. In addition, corolla abscission significantly increased the pollen deposition and seed setting rate. Our results demonstrate that nectar robbing contributes to enhancing seed production and positively and indirectly impacts the reproductive success of S. officinale. This mechanism involved the movement of anthers and indirect participation by nectar robbers, which was rarely investigated. Considering the multiple consequences of nectar robbing, understanding the impact of nectar robbers on plant reproduction is essential to comprehend the evolutionary importance of relationships between plants and their visitors.

Positive or negative anteromedial cortical support of unstable pertrochanteric femoral fractures: A finite element analysis study.

OBJECTIVE: To explore the mechanical stability of unstable pertrochanteric fractures with proximal femoral nail anti-rotation (PFNA-Ⅱ) fixation under different anteromedial cortical support reduction patterns. METHODS: A 65-year-old healthy male volunteer with no history of hip or systemic disease was recruited. High-resolution computed tomography images with a slice thickness of 0.67 mm of his right femur were obtained. These images were used to establish three-dimensional (3D) models. The lesser trochanter and lateral femoral wall were cut off to create an AO Foundation/Orthopaedic Trauma Association type 31-A2 unstable pertrochanteric fracture model. PFNA-Ⅱ was used to simulate fixation. Nine different fracture reduction patterns, which included positive-positive, positive-neutral, positive-negative, neutral-positive, neutral-neutral, neutral-negative, negative-positive, negative-neutral, and negative-negative cortical support reductions, were simulated. A load of 700 N was applied to simulate a 70 kg elderly patient standing on one leg. The models were subjected to finite element analysis. The displacement and von Mises stress distributions were analyzed. RESULTS: The positive-positive cortical support reduction pattern showed minimal stress and the negative-negative reduction pattern showed maximal stress on the intramedullary nail. The stress was mainly concentrated at the junction of the helical blade and the main nail and at the site of the lateral aspect of the insertion point of the nail in the great trochanter. The positive cortical support reduction patterns seemed to have smaller maximum displacements of the entire fragment-implant assembly and relative displacements between the head-neck and shaft fragments. Meanwhile, the negative reduction patterns seemed to have larger displacements. CONCLUSION: The positive-positive support reduction pattern showed better mechanical stability for unstable pertrochanteric fractures. The negative-negative support reduction pattern was prone to fixation failure and should be avoided during an operation.

Enhanced response to PD-L1 silencing by modulation of TME via balancing glucose metabolism and robust co-delivery of siRNA/Resveratrol with dual-responsive polyplexes.

Since cellular metabolism reprogramming is one of the crucial hallmarks of tumor, glucose metabolic pathways are emerging as an important target for modulating immunosuppressive tumor microenvironment (TME) in favor of anti-PD-L1 therapy. Aiming at boosting immune response by modulation immunosuppressive TME via balancing the glycolysis and mitochondrial oxidative phosphorylation (OXPHOS) of tumor cells, we developed a dual-responsive mPEG-PLA-PHis-ss-PEI polyplexes (DRP/Res/siP) for robust co-delivery of PD-L1 siRNA and resveratrol (Res). Isothermal titration calorimetry confirmed the non-electrostatic interactions between PD-L1 siRNA and PHis block of the copolymer, which contributed to the efficient and synchronized release of siRNA with Res in response to the acidic and reductive environment by destabilizing the siRNA polyplexes. The extracellular acidification rate (ECAR) and the oxygen consumption rate (OCR) as well as some key enzymes involved in glycolysis and mitochondrial OXPHOS pathways were determined to quantify the glucose metabolism balance. Effective downregulation of glycolysis and upregulation of mitochondrial OXPHOS were observed in the tumor cells treated with DRP/Res/siP, leading to remarkably reduced lactate production and glucose consumption. In vivo anti-tumor results showed that upregulation of mitochondrial OXPHOS pathways not only significantly promoted CD8+ and CD4+ T cells infiltration, IFN-γ secretion but also significantly suppressed the Treg cells and MDSCs at the same glycolysis level, resulting in superior anti-tumor effect in combination with PD-L1 silencing. Our findings indicate that balancing glucose metabolic pathways of glycolysis and mitochondrial OXPHOS provides a more reliable immune boosting strategy to PD-L1 silencing than exclusive glycolysis inhibition.

Quantitative vibration perception threshold in assessing diabetic polyneuropathy: Should the cut-off value be adjusted for Chinese individuals with type 2 diabetes?

AIMS/INTRODUCTION: To examine the performance and identify the optimal threshold of vibration perception threshold (VPT) for diagnosing diabetic polyneuropathy (DPN) in a Chinese population according to multiple definitions of DPN as gold standards. MATERIALS AND METHODS: VPT was determined in 421 Chinese individuals with type 2 diabetes, who simultaneously completed a questionnaire of neuropathic symptoms, and underwent the assessment of signs of peripheral neuropathy and electromyography tests. Three definitions of DPN (i.e., clinician-diagnosed DPN, abnormal nerve conduction and confirmed DPN) were taken as reference gold standards. RESULTS: Vibration perception threshold was a specific measure for all three groups of DPN outcomes, with the highest specificity noted for clinician-diagnosed DPN (85.1%). The specificity for abnormal nerve conduction and confirmed DPN was 77.0 and 76.6%, respectively. The sensitivity of VPT was 67.0% for clinician-diagnosed DPN, 66.5% for abnormal nerve conduction and 67.2% for confirmed DPN. The optimal cut-off threshold for abnormal nerve conduction, as well as confirmed DPN, was VPT >14.9 V. The specificity and sensitivity of VPT >14.9 V as the cut-off value for clinician-diagnosed DPN were 85.6 and 66.2%, respectively. When taking clinician-diagnosed DPN as the gold standard, the performance of VPT for diagnosing DPN was best with an area under the curve value of 0.804. CONCLUSIONS: VPT measured using the neurothesiometer had relatively high specificity and best performance for diagnosing DPN when clinician-diagnosed DPN rather than abnormal nerve conduction was taken as the gold standard in a Chinese population. A VPT value of ≥15 V might be equally applicable for diagnosing DPN in a Chinese population.

[Successful Treatment of Refractory Ischemic Diabetic Foot Ulcers by Combination Therapy of Autologous Platelet-rich Gel and Topical ß Adrenergic Receptor Blocker: a Case Report].

HISTORY AND CLINICAL FINDINGS: A 76 year-old woman with 8-year history of diabetes mellitus and hypertension was admitted with gangrene of left great toe, 3rd, 4th and 5th toes. Twenty months ago, She started to receive hemodialysis due to end-stage renal disease. She did not have any history of reactive airway disease nor bradycardia that would contraindicate the use of topical beta-blocker. The X-ray of left lower limb and foot showed calcification of left superficial femoral artery, popliteal artery, anterior tibial artery, posterior tibial artery, dorsal foot artery and digital artery, as well as osteolytic destruction at distal end of metatarsal bone, and lateral dislocation of the 4th and 5th toes. Color Doppler ultrasound of bilateral lower extremity arteries showed obvious calcification of bilateral superficial femoral arteries, thrombosis of left popliteal artery, severe stenosis of left anterior tibial artery, occlusion of left posterior tibial artery, right anterior tibial artery and posterior tibial artery. Computed tomographic angiography (CTA) of bilateral lower limb arteries revealed moderate stenosis of left superficial femoral artery, occlusion of left popliteal artery, left posterior tibial artery and dorsal pedal artery, occulusion of right posterior tibial artery, but right dorsal pedal artery was visible. DIAGNOSIS, TREATMENT AND FOLLOW-UP: Diagnosis of diabetic foot (left, grade 4) and diabetic lower extremity arterial occlusion (left, stage 4) was made. Based on multidisciplinary team (MDT) discussion, the patient was unable to undergo vascular bypass surgery, and left lower extermity amputation also was not suitable because of right atrial thrombosis. Therefore, conservative treatment was recommended. The specific scheme used clopidogrel for antiplatelet agglutination, Low Molecular Weight Heparin (Clexane) and warfarin for anticoagulation, lipo-alprostadil for vasodilation, as well as local debridement and ultrasonic debridement. The treatments were given for up to 9 weeks, but with no significant clinical response. So the patient was treated with vacuum-assisted closure and autologous platelet-rich gel therapy for the next 7 weeks, then applied with 1 drop of timolol maleate 0.5% ophthalmic solution per cm 2 wound area every other day for another 6 weeks, the wound rapidly healed and re-epithelialized basically. The follow-up for 5 weeks showed that the wound healed completely without any discomfort. No side effect was found.