Objective To investigate the effects of low dose of extracellular histone on endothelial cells in infectious diseases such as sepsis. Methods The endothelial cells were treated with 10 μg/mL recombinant human histone H3/H4 complex in replacement of calf thymus histones (CTH) for various periods of time, and the morphology changes and the viability of the endothelial cells were recorded. In addition, flow cytometry was applied to identify the characteristics of endothelial cells and enzyme-linked immunosorbnent assay (ELISA) was used to detect the extracellular histones level in endothelial cells culture. Results The low dose of CTH could continuously induce endothelial cells death, cell morphological changes and function loss, which was reproduced by 10 μg/mL recombinant histone H3/H4 complex. Results of histones quantitation showed that histone can cause a series of intracellular reactions in a short period of time. Conclusions It is showed that 10 μg/mL H3/H4 can induce the toxicity in infectious disease and this level of the dose is a lower than those used in previous studies and more close to the pathological conditions.

ObjectivesTo explore the clinical significance of tyrosine kinase receptor RON mRNA expression and it's splicing variant in bladder tumors. Methods Sixty-three cases of transitional cell carcinoma of the bladder (TCCB), including 30 cases of pathological grade I , 15 cases of pathological grade Ⅱ and 18 cases of pathological grade Ⅲ (44 cases of clinical stage Tis + T1, 15 cases of T2 + T3 +T4), 7 inverted papilloma of the bladder ( IPB), 9 cases of papillary urothelial neoplasm of low malignant potential (PUNLMP) and 12 cases of normalbladder mucosa RT-PCR was employed with the internal standard (GAPDHmRNA) to detect the expression of RON mRNA. PCR and direct sequencing was then utilized to identify the potential RON mRNA splicing variants. Finally, the variants' positive rates of expression were analyzed among the different tissues, diverse TCCB pathological grades and clinical stages. ResultsThe expression levels of RON mRNA/GAPDH mRNA among TCCB, IPB, PUNLMP and normal control were 4. 9 × 10-3 ( 1. 8 × 10-3-1.0 × 10-2 ), 3. 8 × 10-3 (2. 4 × 10-3-1.7 × 10-2 ), 4. 9 ×10 -3 ( 1.7 × 10 -3-1.1 × 10 -2 ) and 1.0 × 10-3 (4. 5 × 10-4-2. 8 × 10-3 ) respectively. The difference had a statistical significance (x2K-W = 17. 278 ,P ＜0. 05 ). The expression levels among pathological grade I, Ⅱ andⅢ were 3.7 × 10-3( 1.3 × 10-3-7.5 × 10-3) , 4. 9 × 10-3(1.9 × 10-3-1.1 × 10-2) and 8.9 × 10-3(2. 7 ×10 -3-8.0 × 10 -2 ) respectively. The erpression levels among the clinical stage Tis + T1 and T2 + T3 + T4were 3.5 × 10-3 ( 1.2 × 10 -3-7. 7 × 10-3 ) and 9. 7 × 10 -3 ( 2. 9 × 10-3-5. 3 × 10-2 ). The differences between expression levels were of statistical significance among the different pathological grades ( x2k-W =7. 341, P ＜0. 05 ) and clinical stages ( Z = - 2. 306, P ＜ 0. 05 ). The positive rates of exon 11deletion(E11△) in TCCB, IPB and PUNLMP were 71％ (45/63), 57％ (4/7) and 67％ (6/9) respectively, andthe total positive rate in bladder tumor tissues was 70％. Meanwhile, expression of the novel RON variant wesnot detected in the normalbladder mucosa. The positive expression rate of E1 1△ has no significant correlationamong the different clinical pathological tissues (x2 = 0. 620, P ＞ 0. 05 ). There was no statistical significancein expression positive rate between different pathological grades ( Z =0. 221, P ＞0. 05 ) and clinical stages( Z = 0. 538, P ＞ 0. 05) as well. A novel RON splice variant, deletion of RON exon 11 3 476 - 3 539 ( E3476 -3539△) was fond in the pathological tissue. The positive expression rates of E3 476 -3 539 in TCCB,IPB and PUNLMP were 57％ (36/63), 43％ (3/7) and 56％ (5/9) respectively, and the total positive expression rate was 56％ (44/79). The positive rates of E3 476 -3 539△ in pathological grade I , Ⅱ and Ⅲ were 40％ ( 12/30), 67％ (10/15) and 78％ (14/18), and it's positive rates in clinical stage Tis +T1and T2 +T3 + T4 were 48％ (21/44) and 80％ (12/15). The differences in each group had significantly statistical significance ( Z = 7. 285, 5. 041, P ＜ 0. 05 ) . However, the positive rates amongdifferent pathological tissues had no significance (x2 = 0. 517, P ＞ 0. 05 ). Conclusions The expression level of RON mRNA is significantly associated with histological grading and clinical stage. RON may play an important role in the progression ofTCCB. Compared with the normal control, the increased RON variant expression may contribute to the carcinogenesis of the bladder tumor.

Objective To investigate the change of extracellular histone level as well as the mecha-nism of the cytotoxicity of extracellular histones on vascular endothelial cell in sepsis. Methods Septic chil-dren admitted to PICU in Shanghai Children′s Medical Center between January 2010 and December 2014 were included in the present study. According to the diagnostic criteria of sepsis,the patients were divided into the sepsis group(51 cases) and the severe sepsis group(79 cases),with healthy children as the control group (108 cases). Patients in the severe sepsis group were further divided into the survival group(45 cases) and the non-survival group ( 34 cases ) based on 28-day mortality. The plasma concentration of extracellular histones in these children was determined and its correlation with the severity of sepsis was analyzed. Human umbilical vein endothelial cells(HUVECs) were incubated with calf thymus histone(CTH) at various con-centrations(0,50,100,200 and 300μg/ml) or different time periods(200μg/ml,0,5,15,30,45 and 60 mi-nutes) . The treated cells were subject to flow cytometer to measure the cell survival rate and scanning/trans-mission electron microscopy to observe their morphological changes. Western blot was used to detect the ex-pression of IκB and phosphor-p38/p38 in nuclear factor ( NF )-κB and mitogen-activated protein kinase ( MAPK) signaling pathways,while ELISA was used to determine the levels of tumor necrosis factor-α and interleukin-6. Results The levels of circulating histones in the septic children(2. 29 ± 1. 00) and severe sep-tic children ( 19. 17 ± 10. 20 ) were significantly higher than that of healthy controls ( 0. 23 ± 0. 26 ) ( P <0. 001),and the histone levels in the severe septic children were even higher(P<0. 001). Among the chil-dren diagnosed as severe sepsis,the level of circulating histones in the non-survivors was significantly higher than that in the survivors(29. 47 ± 5. 99 vs. 10. 94 ± 2. 68,P<0. 001). The survival rate of HUVEC gradually decreased along with the increase of CTH concentration or the treatment period in vitro. Data from electron microscopy showed that CTH treatment could directly disrupt the plasma membrane of HUVEC. Histones could also activate NF-κB and MAPK pathways,leading to the release of large amount of tumor necrosis fac-tor-α and interleukin-6. Conclusion The levels of extracellular histones in the septic children are correlated with the severity of sepsis. CTH can induce HUVEC death in a dose-and time-dependent manner. Extracellu-lar histone-induced endothelial dysfunction may mediate the progression of sepsis and such cytotoxicity might be due to the destruction of endothelial cell membranes and activation of the NF-κB and MAPK signaling pathways.

BACKGROUND:Currently, literatures about autologous vein transplantation are few, and the research on the effect of different parts of autologous vein transplantation are not found yet. OBJECTIVE:To summarize the experiences of establishing the fistula using autologous vein transplantation so as to investigate the method of improving the success rate of surgery. METHODS:We analyzed retrospectively the data of 40 cases of establishing the fistula using autologous vein transplantation, and then compared the successful rate of autologous vein transplantation fistula, blood flow and operating time, thereby analyzing the influence of diabetes melitus on the successful rate of autologous vein transplantation fistula. RESULTS AND CONCLUSION:The successful rates of autologous vein transplantation fistula at different parts ranging from high to low were as folows: the cephalic vein, great saphenous vein, basilic vein and smal saphenous vein. Blood flow of the upper limb for vein transplantation fistula was obviously higher than that of the lower limb (P < 0.05). The operating time of autologous vein transplantation fistula was longer in the upper limbs than in the lower limbs (P < 0.01). For patients with diabetes melitus, the successful rate of autologous vein transplantation was markedly lower than those with no diabetes melitus (P < 0.01). For the hemodialysis patients with poor upper limb superficial vein, autologous vein transplantation is a better way of establishing the vascular access. Vein transplantation of the upper limbs is better than that of the lower limbs in success rate and operating time. Autologous vein transplantation fistula is not suitable for the patients with diabetes melitus.

Objective To discuss experiences in reconstruction of severe tibial shaft fractures by using different flaps and external fixations.Methods The study involved 190 patients with type GustiloⅢB Ⅲ C (160 patients) and GustiloⅢC (30 patients) tibial shaft fractures treated from 1990 to 2007.There were 169 males and 21 females,at average age of 42.5 years.The injury causes included traffic accidents in 132 patients,machine accidents in 32 and stone smashing in 26.The management procedure consisted of administration of antibiotics,serial debridement and different flap grafting (including free thoracoumbilical flaps in 20 patients,sural neurocutaneous vascular flaps in 108,saphenous neurocutane ous vascular flaps in 12,superficial peroneal neurocutaneous flap in two,fasciocutaneous flaps in 26 and gastrocnemius muscular flaps in 22) and different external fixators (half-ring fixators in 84 patients,unilateral axial dynamic fixators in 12,AO fixators in 10,Weifang fixators in 42 and hybrid fixators in 40).The average follow-up was 7.3 years.Results All flaps survived.Of all,186 patients obtained fracture healing,with mean fracture healing time varying in different patients treated with different external fixators:7.5 months for 84 patients treated with half-ring fixators,11.2 months for eight with unilateral axial dynamic fixators,8.5 months for 12 with AO fixators ,8.1 months for 42 with Weifang fixators and 7.8 months for 40 with assembly fixators.Except for half-ring fixation,the other fixators needed necessary bone graft.Four patients treated with unilateral axial dynamic fixators resulted in nonunion due to osteo myelitis.The latest follow up showed that the function of the ankle and knee was normal,with no pain.Conclusion Combination of half-ring external fixators with various flaps provides good method for treatment of Gustilo ⅢB and ⅢC tibial shaft fractures.

Objective To develop a magnetic resonance imaging (MRI) system for pelagic definitive treatment platforms, large support ships and ocean vessels.Methods The system was designed based on normal electromagnetic MRI technology, which was composed of six parts of gradient system, magnet system, coil system, radiofrequency system, spectrum control system and imaging system.The magnet system was developed with C-shaped open magnet,five systems were designed such as a super-stability excitation system, a super-sensitivity signal detection system, a magnetic filed dissipation precision control system, a shipborne sensitive equipment vibration control system and a software system, and a set of technical detection standards was proposed.Results The developed MRI system realized mobile imaging,gained advantages in imaging velocity,resolution,magnetic field dissipation control,ship bump resistance and environmental suitability,and thus could be used in marine mobile conditions. Conclusion The developed MRI system meets the requirements of pelagic medical treatment, and contributes to improving pelagic medical support ability of Chinese navy.

OBJECTIVETo investigate the presence of Cosmc gene mutation in children with Henoch-Schönlein purpura (HSP) and the association between Cosmc gene mutation and the susceptibility to HSP.

MESULTSEighty-four children who were diagnosed with HSP between March 2014 and December 2015 were selected as the HSP group. Fifty-eight healthy volunteers matched for age and sex were enrolled as the control group. Fasting venous blood (5 mL) from the two groups was collected in EDTA anticoagulated tubes, followed by the isolation of peripheral blood mononuclear cells (PBMCs) through density gradient centrifugation. Genomic DNA was extracted from PBMCs according to the manufacturer's protocol, and the whole exon region of Cosmc gene was amplified by touch-down polymerase chain reaction (touch-down PCR). The PCR products were identified by 1% agarose gel and sequenced in order to further examine the association between Cosmc gene mutation and the susceptibility to HSP.

RESULTSSequencing results showed two mutations (c.393T>A and c.72A>G) of Cosmc gene in children with HSP. There were no significant differences in the genotype and allele frequencies at the two loci between the HSP and control groups, and this distribution was not associated with sex.

CONCLUSIONSThe mutations c.393T>A and c.72A>G in the exon region of Cosmc gene in children with HSP are not associated with the onset of HSP.

Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Molecular Chaperones ; genetics ; Mutation ; Purpura, Schoenlein-Henoch ; etiology ; genetics

Objective:To analyze the clinical efficacy of treatment of cement dislodgement after vertebral augmentation for osteoporotic vertebral fractures.Methods:A retrospective study was conducted to analyze the data of 13 patients who had been treated at Department of Orthopaedics, The First Affiliated Hospital of Soochow University for cement dislodgement after vertebral augmentation for osteoporotic vertebral fractures from July 2013 to July 2022. There were 4 males and 9 females, with an average age of (76.5±8.6) years and a T value of bone mineral density of -3.3±0.6. By the CT and MRI features of cement dislodgement, their conditions fell in 4 types: cement loosening in situ (4 cases), anterior cement moving (6 cases), anterior cement moving with posterior bone mass moving (2 cases), and posterior cement moving (1 case). They were treated by percutaneous vertebroplasty (3 cases), pedicle screw fixation combined with bone graft fusion and decompression (7 cases), and conservative therapy (3 cases). The curative effects for surgical patients were evaluated by comparing their visual analogue scale (VAS), Oswestry dysfunction index (ODI) and cobb angle of kyphosis at preoperation, 1 week and 1 month postoperation, and the last follow-up, and Frankel grading for nerve injury as well. The curative effects for patients undergoing conservative treatment were evaluated by observing their symptoms.Results:This cohort was followed up for 7 (5, 12) months after treatment. The VAS scores [5.0 (4.0, 5.0) points, 3.0 (2.0, 3.0) points, and 3.0 (2.0, 3.0) points] in the 10 surgical patients at 1 week and 1 month postoperation and the last follow-up were significantly improved compared with the preoperative value [8.5 (8.0, 9.0) points] ( P<0.05); the VAS scores at 1 month postoperation and the last follow-up were also significantly improved compared with that at 1 week postoperation ( P < 0.05), but there was no significant difference between the last follow-up and 1 month postoperation ( P > 0.05). The ODIs (50.6%±4.2%, 37.8%±4.5%, and 29.3%±5.6%) in the 10 surgical patients at 1 week and 1 month postoperation and the last follow-up were significantly improved compared with the preoperative value (93.2%±3.6%), showing significant differences in pairwise comparisons ( P<0.05). The cobb angles [10.0 (9.0, 11.0)°, 9.0 (9.0, 11.0)°, and 10.0 (9.0, 12.0)°] in the 10 surgical patients at 1 week and 1 month postoperation and the last follow-up were significantly improved compared with the preoperative value [12.5 (11.0, 14.0)°] ( P<0.05) , but there was no statistically significant difference between the time points after operation ( P>0.05). The Frankel grading was significantly improved in the 6 patients with nerve injury after operation. Of the 3 patients undergoing conservative treatment, the symptoms were cured in one, showed no change during follow-up in one, and aggravated in one. Conclusion:Surgical treatment can significantly relieve pain, improve spinal dysfunction and repair nerve injury in patients with bone cement dislodgement after vertebral augmentation.

BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice.MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH 2 O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected.Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

Objective:To explore the effects of accelerated rehabilitation surgery (ERAS) on visual analogue scale (VAS) and serum immunoglobulin in children undergoing low-temperature plasma radiofrequency tonsillectomy.Methods:A prospective study was conducted on 200 children who underwent bilateral tonsillectomy with low-temperature plasma radiofrequency from January 2022 to April 2023 at the Sixth Affiliated Hospital of South China University of Technology. They were randomly divided into an observation group ( n=100) and a control group ( n=100) using a random number table method. The control group received traditional perioperative treatment, while the observation group received perioperative treatment under ERAS mode. We compared the postoperative conditions, pain VAS at different time points, and incidence of complications between two groups, as well as serum immunoglobulins and inflammatory markers before and 2 days after surgery. Results:The observation group had shorter postoperative pseudo film detachment time time, white membrane formation time, dietary recovery time, and hospital stay than the control group (all P<0.05); The VAS of the observation group was lower than that of the control group at different time points after surgery (all P<0.05); Two days after surgery, the levels of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in both groups decreased, with the observation group showing a smaller decrease than the control group (all P<0.05); Two days after surgery, the levels of interleukin-8 (IL-8), interleukin-6 (IL-6), and C-reactive protein (CRP) significantly increased in both groups, with the observation group showing a lower increase than the control group (all P<0.05); The total incidence of complications in the observation group was 5.00%(5/100), significantly lower than the control group′s 13.00%(13/100, P<0.05). Conclusions:After undergoing ERAS intervention during the perioperative period, children undergoing low-temperature plasma tonsillectomy can accelerate their recovery process, shorten hospital stay, reduce pain VAS and inflammatory indicators, and reduce the impact on their serum immunoglobulin levels.