OBJECTIVES: To estimate postmortem interval (PMI) by analyzing the protein changes in skeletal muscle tissues with the protein chip technology combined with multivariate analysis methods. METHODS: Rats were sacrificed for cervical dislocation and placed at 16 ℃. Water-soluble proteins in skeletal muscles were extracted at 10 time points (0 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d, 7 d, 8 d and 9 d) after death. Protein expression profile data with relative molecular mass of 14 000-230 000 were obtained. Principal component analysis (PCA) and orthogonal partial least squares (OPLS) were used for data analysis. Fisher discriminant model and back propagation (BP) neural network model were constructed to classify and preliminarily estimate the PMI. In addition, the protein expression profiles data of human skeletal muscles at different time points after death were collected, and the relationship between them and PMI was analyzed by heat map and cluster analysis. RESULTS: The protein peak of rat skeletal muscle changed with PMI. The result of PCA combined with OPLS discriminant analysis showed statistical significance in groups with different time points (P<0.05) except 6 d, 7 d and 8 d after death. By Fisher discriminant analysis, the accuracy of internal cross-validation was 71.4% and the accuracy of external validation was 66.7%. The BP neural network model classification and preliminary estimation results showed the accuracy of internal cross-validation was 98.2%, and the accuracy of external validation was 95.8%. There was a significant difference in protein expression between 4 d and 25 h after death by the cluster analysis of the human skeletal muscle samples. CONCLUSIONS The protein chip technology can quickly, accurately and repeatedly obtain water-soluble protein expression profiles in rats' and human skeletal muscles with the relative molecular mass of 14 000-230 000 at different time points postmortem. The establishment of multiple PMI estimation models based on multivariate analysis can provide a new idea and method for PMI estimation.
Animals ; Humans ; Rats ; Multivariate Analysis ; Postmortem Changes ; Protein Array Analysis ; Technology

Objective:To explore the efficacy of the proximal tibial lateral locking plate with rafting screws combined with Jail screws in the treatment of collapse fracture of the lateral tibial plateau.Methods:A retrospective case series study was performed on clinical data of 36 patients with collapse fracture of the lateral tibial plateau admitted to Suqian Hospital of Nanjing Drum Tower Hospital Group from January 2016 to January 2020, including 19 males and 17 females, aged 34-68 years [(48.6±5.8)years]. Schatzker classification was type II in 28 patients and type III in 8 patients. All patients were treated using the proximal tibial lateral locking plate with raft screws combined with Jail screws. The operation time, intraoperative blood loss and fracture healing were detected. The distance of articular surface collapse of the tibial plateau, posterior inclination angle (PSA) of the tibial plateau, tibial plateau varus angle (TPVA) and Rasmussen anatomical score were compared before operation and at day 2 and 1 year after operation. The Hospital for Special Surgery (HSS) score was used to evaluate knee function at day 2 and 1 year after operation. Complications were also recorded.Results:All patients were followed up for 12-32 months [(19.5±3.1)months]. The operation time was 56-82 minutes [(68.5±9.1)minutes]. The intraoperative blood loss was 40-100 ml [(75.6±10.2)ml]. The fracture was clinically healed by first-stage, with the healing time of 8 to 15 weeks [(12.5±1.3)weeks]. The distance of articular surface collapse of the tibial plateau was improved from (15.5±4.2)mm before operation to (0.7±0.3)mm at day 2 after operation and (1.0±0.2)mm at 1 year after operation (all P<0.01). The PSA was improved from (21.2±2.1)° before operation to (8.9±0.8)° at day 2 after operation and (9.2±0.6)° at 1 year after operation (all P<0.01). The TPA was improved from (100.2±3.7)° before operation to (88.9±1.8)° at day 2 after operation and (87.2±1.6)° at 1 year after operation (all P<0.05). The Rasmussen anatomical score changed from (7.8±1.8)points before operation to (17.1±0.9)points at day 2 after operation and (16.3±0.7)points at 1 year after operation (all P<0.01). There were no significant difference in the distance of articular surface collapse of the tibial plateau, PSA, TPVA and Rasmussen anatomical score at day 2 and 1 year after operation (all P>0.05). The HSS score was (92.8±3.2)points at 1 year after operation, significantly higher than (74.8±3.5) points at day 2 after operation ( P<0.01). Two patients sustained superficial wound infection after operation, which healed after debridement and dressing change. No deep infection occurred. Conclusion:For patients with collapse fracture of the lateral tibial plateau, the proximal tibial lateral locking plate with rafting screws combined with Jail screws can effectively prevent secondary collapse of the articular surface and obtain satisfactory anatomical reduction, good functional recovery and few postoperative complications.

Objective:To compare the clinical effects of minimally invasive reduction through a bone tunnel combined with Jail screwing and those of posterolateral locking plating in the treatment of simple posterolateral tibial plateau fractures.Methods:A retrospective analysis was conducted of the data of 48 patients who had been operatively treated and completely followed up at Department of Orthopedics, Suqian Hospital of Nanjing Drum Tower Hospital Group for simple posterolateral tibial plateau fractures from October 2016 to October 2020. There were 26 males and 22 females, aged from 35 to 68 years. They were divided into a minimally invasive group (25 cases subjected to minimally invasive reduction through a bone tunnel combined with Jail screwing) and an incision group (23 cases subjected to posterolateral locking plating) according to their surgical methods. The operation time, incision length, intraoperative blood loss, fracture healing time, cumulative fluoroscopy time, hospital stay and posterior inclination angles of the tibial plateau and Hospital for Special Surgery (HSS) knee function scores at 1, 3, 6, 9, and 12 months after operation were compared between the 2 groups. Complications in the 2 groups of patients were recorded.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). The 48 patients were followed up for 12 to 36 months (average 16.5 months). The minimally invasive group was significantly better than the incision group in operation time [(42.6±9.1) min versus (65.7±11.5) min], incision length [(4.0±0.4) cm versus (15.0±1.5) cm], intraoperative blood loss[(22.6±5.8) mL versus (31.5±8.8) mL], hospital stay [(7.6±1.4) d versus (11.1±2.4) d], and HSS score one month after operation [(84.8±1.9) points versus (72.9±4.1) points], but the cumulative fluoroscopy time in the incision group [(4.1±1.4) s]was significantly less than that in the minimally invasive group [(22.3±4.2) s] ( P<0.05). There were no significant differences in fracture healing time, HSS scores at 3, 6, 9, or 12 months after operation, or posterior inclination angle of the tibial plateau between the 2 groups ( P>0.05). There were no such complications as wound infection, vascular injury, internal fixation failure, nonunion or malunion of fractures in either of the 2 groups. Two cases in the incision group presented with symptoms of common peroneal nerve injury but recovered 3 months after operation. Conclusions:Although both minimally invasive reduction through a bone tunnel combined with Jail screwing and posterolateral locking plating can achieve satisfactory outcomes in the treatment of simple posterolateral tibial plateau fractures, the minimally invasive technique is preferable because it shows the advantages of a smaller incision, less bleeding, shorter operation time, a lower operation risk, quicker postoperative recovery and shorter hospital stay.

BACKGROUND: Perioperative treatment has become an increasingly important aspect of the management of patients with non-small cell lung cancer (NSCLC). Small-scale clinical studies performed in recent years have shown improvements in the major pathological remission rate after neoadjuvant therapy, suggesting that it will soon become an important part of NSCLC treatment. Nevertheless, neoadjuvant immunotherapy may be accompanied by serious adverse reactions that lead to delay or cancelation of surgery, additional illness, and even death, and have therefore attracted much attention. The purpose of the clinical recommendations is to form a diagnosis and treatment plan suitable for the current domestic medical situation for the immune-related adverse event (irAE). METHODS: This recommendation is composed of experts in thoracic surgery, oncologists, thoracic medicine and irAE related departments (gastroenterology, respirology, cardiology, infectious medicine, hematology, endocrinology, rheumatology, neurology, dermatology, emergency section) to jointly complete the formulation. Experts make full reference to the irAE guidelines, large-scale clinical research data published by thoracic surgery, and the clinical experience of domestic doctors and publicly published cases, and repeated discussions in multiple disciplines to form this recommendation for perioperative irAE. RESULTS: This clinical recommendation covers the whole process of prevention, evaluation, examination, treatment and monitoring related to irAE, so as to guide the clinical work comprehensively and effectively. CONCLUSIONS Perioperative irAE management is an important part of immune perioperative treatment of lung cancer. With the continuous development of immune perioperative treatment, more research is needed in the future to optimize the diagnosis and treatment of perioperative irAE.

A new electrochemical sensor for organophosphate pesticide(methyl-paraoxon)detection based on bifunctional cerium oxide(CeO2)nanozyme is here reported for the first time.Methyl-paraoxon was degraded into p-nitrophenol by using CeO2 with phosphatase mimicking activity.The CeO2 nanozyme-modified electrode was then synthesized to detect p-nitrophenol.Cyclic voltammetry was applied to investigate the electrochemical behavior of the modified electrode,which indicates that the signal enhancement effect may attribute to the coating of CeO2 nanozyme.The current research also studied and discussed the main parameters affecting the analytical signal,including accumulation potential,accumulation time,and pH.Under the optimum conditions,the present method provided a wider linear range from 0.1 to 100 μmol/L for methyl-paraoxon with a detection limit of 0.06 μmol/L.To validate the proof of concept,the electrochemical sensor was then successfully applied for the determination of methyl-paraoxon in three herb samples,i.e.,Coix lacryma-jobi,Adenophora stricta and Semen nelum-binis.Our findings may provide new insights into the application of bifunctional nanozyme in electro-chemical detection of organophosphorus pesticide.

Transposable elements (TEs) are a major determinant of eukaryotic genome size. The collective properties of a genomic TE community reveal the history of TE/host evolutionary dynamics and impact present-day host structure and function, from genome to organism levels. In rare cases, TE community/genome size has greatly expanded in animals, associated with increased cell size and changes to anatomy and physiology. Here, we characterize the TE landscape of the genome and transcriptome in an amphibian with a giant genome—the caecilian Ichthyophis bannanicus, which we show has a genome size of 12.2 Gb. Amphibians are an important model sys-tem because the clade includes independent cases of genomic gigantism. The I. bannanicus genome differs compositionally from other giant amphibian genomes, but shares a low rate of ectopic recombination-mediated deletion. We examine TE activity using expression and divergence plots;TEs account for 15％of somatic transcription, and most superfamilies appear active. We quantify TE diversity in the caecilian, as well as other vertebrates with a range of genome sizes, using diver-sity indices commonly applied in community ecology. We synthesize previous models that integrate TE abundance, diversity, and activity, and test whether the caecilian meets model predictions for genomes with high TE abundance. We propose thorough, consistent characterization of TEs to strengthen future comparative analyses. Such analyses will ultimately be required to reveal whether the divergent TE assemblages found across convergent gigantic genomes reflect fundamental shared features of TE/host genome evolutionary dynamics.

Single-cell genomics provides substantial resources for dissecting cellular heterogeneity and cancer evolution. Unfortunately, classical DNA amplification-based methods have low throughput and introduce coverage bias during sample preamplification. We developed a single-cell DNA library preparation method without preamplification in nanolitre scale (scDPN) to address these issues. The method achieved a throughput of up to 1800 cells per run for copy number variation (CNV) detection. Also, our approach demonstrated a lower level of amplification bias and noise than the multiple displacement amplification (MDA) method and showed high sensitivity and accuracy for cell line and tumor tissue evaluation. We used this approach to profile the tumor clones in paired primary and relapsed tumor samples of hepato-cellular carcinoma (HCC). We identified three clonal subpopulations with a multitude of aneuploid alterations across the genome. Furthermore, we observed that a minor clone of the primary tumor containing additional alterations in chro-mosomes 1q, 10q, and 14q developed into the dominant clone in the recurrent tumor, indicating clonal selection during recurrence in HCC. Overall, this approach provides a comprehensive and scalable solution to understand genome hetero-geneity and evolution.

Objective To evaluate the clinical significance of serum thrombospondin-1 (TSP-1) in systemic lupus erythematosus (SLE), and explore its possible involvement in SLE pathogenesis. Methods One hundred and thirty-eight patients diagnosed with SLE, including 124 cases of females and 14 males, as well as 60 healthy controls were recruited into this study. Enzyme-linked immunosorbent assay (ELISA) was used to determine serum TSP-1 expression level between the two groups. Spearman correlation analysis method was used to analyze the correlation between TSP-1 level of complement 3, complement 4, anti-double-stranded DNA, Rheumatoid factor and systemic lupus erythematosus disease activity index (SLEDAI). Results TSP-1 level in healthy control group was much higher than that in SLE patients. TSP-1 serum levels in SLE patients was positively correlated with complement 3 (r=0.386, P<0.01), complement 4 (r=0.301, P<0.01), and negatively correlated with anti-double-stranded DNA (r=-0.221, P=0.009), RF (r=-0.186, P=0.029) and SLEDAI (r=-0.273, P=0.001). Conclusion TSP-1 may play an immune-regulatory role in the development of SLE by inhibiting inflammation and autoantibody production. It could become a potential therapeutic target for the treatment of SLE.

Objective To compare the outcomes of arthroscopic single-row and suture-bridge repair of medium-sized rotator cuff tears through clinical and MRI assessment.Methods Forty-five patients with medium-sized rotator cuff tears who underwent arthroscopic repair using single-row (SR,n=29) or suture-bridge (SB,n=26) technique between July 2014 to June 2015 in our hospital,were retrospectively enrolled in this study.All surgeries were performed by the same senior doctor.The functional outcomes were assessed using the rating scale of the University of California at Los Angeles (UCLA),American Shoulder and Elbow Surgeons shoulder index (ASES),Fudan University Shoulder Score (FUSS),visual analog pain scale score (VAS),and range of motion (ROM) before the operation and at the last follow-up.MRI examination was performed at the final follow-up.Results Finally 50 patients were followed up successfully,24 in SB group and 26 in SR group.There were no significant differences between the two groups in terms of age,sex,follow-up duration,and the affected side.Before the operation,no significant differences were observed between the 2 groups in all the measurements.After more than 1-year follow-up,significant improvement was found in all measurements for both groups,but without significant differences between them.MRI examination showed no re-tears in either group.According to the Sugaya's classification,there were more type Ⅰ patients in SB group than SR group (83.3％ vs 61.5％),but the difference was not significant.Conclusion Arthroscopic suture-bridge repairing of medium-sized rotator cuff tears results in safe and good early clinical outcomes.However,compared with the single-row technique,there was no significant advantage.Although the MRI examination showed a better result of rotator cuff healing in the early stage,its long-term outcomes need further studying.

Background Oxidative stress is a pathophysiological process of retina,so it is very important to explore a protective way against retinal oxidative stress.Studies determined that extract of ginkgo biloba (EGb) has antioxidant,anti-apoptosis,anti-thrombosis and anti-inflammatory effects,however,the effect of EGb on human Müller cells in oxidative stress is still below understood.Objective This study was to investigate the protection of EGb against oxidative stress of human retinal Müller cells induced by As2O3 in vitro.Methods Human retinal Müllercell line was cultured in DMEM/F12 with 10％ fetal bovine serum (FBS),2 μmol/L glutamine and 1％ antibiotics.As2O3 solution at the final concentration 5 mg/L was added in the medium for 24 hours to establish oxidative models,and then the EGb with the final concentrations of 5,10 and 20 mg/L was used to cell models for 24 hours,respectively.Cell viability was detected by MTT assay,and reactive oxygen species (ROS) levels in the cells were detected with CM-H2DCFDA fluorescent probe.The relative expression levels of caspase-3 mRNA in cytoplasm and cell nuclei were assayed by reverse-transcription PCR (RT-PCR),and the expressions of Nrf2 protein were quantitatively detected by Western blot.Results Müller cells adhered well 24 hours after cultured.At 6-7 days after culture,Müller cell body was large with abundant cytoplasm and mosaic-like arrangement.However,floating cells were seen after As2 O3 treatment.Cell viability (absorbance) was significantly different among the normal culture group,As2 O3-treated group,As2 O3 + 5 mg/L EGb group,As2 O3 + 10 mg/L EGb group and As2 O3 + 20 mg/L EGb group,with the strongest viability in the normal culture group and the weakest viability in the As2 O3-treated groups (F=163.57,P =0.00).The fluorescence intensity of ROS was the weakest in the normal culture group and the strongest in the As2 O3-treated group and was gradually weakened with the increase of EGb doses,showing a remarkable difference among the groups (F =4 013.61,P =0.00).The relative expression level of caspase-3 mRNA in the cells was gradually reduced with the increase of EGb doses,with a statistically significant difference among the groups (F =2 199.72,P =0.00).In addition,no considerable difference was seen in the expression level of Nrf2 protein (grey scale) in cytoplasm among the groups (F=15.42,P=0.40);while in the nuclei,the expression levels of Nrf2 protein were 100.01 ±0.04,46.59±0.63,54.51 ±0.62,59.93 ±0.17 and 67.60±0.24 in the normal culture group,As2 O3-treated group and As2O3+5 mg/L EGb group,As2O3+10 mg/L EGb group,As2O3+20 mg/L EGb group respectively,with a significant difference among them (F=7 271.72,P=0.00).Conclusions EGb can protect human retinal Müller cells against As2O3-induced damage in a dose-dependant manner by antioxidant and antiapoptotic effects in vitro,and the activities occur primarily in cell nucleus.