Practical Value of Environment-friendly Sample Release Agent Combined with Ultrasound in the Preparation of Pathological Tissue Sections / 中山大学学报(医学科学版)

ObjectiveTo explore the practical value of environment-friendly sample release agent combined with ultrasound in the preparation of pathological tissue sections. MethodsFrom February 2013 to December 2022， 2 518 pathological specimens submitted by Foshan Municipal Hospital of Traditional Chinese Medicine were selected as the study objects. Two samples of the same specimen were randomly divided into two groups： the environment-friendly fast group， in which the pathological tissue sections were made by using the environment-friendly sample release agent combined with ultrasound； and the traditional group， in which formaldehyde， ethanol and xylene were used to make slices in the conventional way. The differences of hematoxylin （HE） staining effect， immunohistochemistry （IHC） staining effect and MDM2 gene detection result of atypical lipomatous tumor/highly differentiated liposarcoma （ALT/WDL） tissue sections between the two groups were compared. Results① The wax of the two groups' pathological tissues was dehydrated well and the tissue hardness was moderate. After HE staining， the sections of the two groups were intact， without cracks and tremor marks， and the contrast between nucleus and cytoplasm was appropriate， with good transparency， uniform staining， and no tissue loss. The excellent rate and score of HE staining in the environmental fast group were higher than those in the traditional group， but the difference was not statistically significant （χ2 = 3.125，P1 = 0.070；t = 0.965，P2 = 0.334）. ②After IHC staining of the two groups of sections， the positive location of the cells was accurate， the staining was specific and uniform， the staining intensity was moderate， the staining sensitivity was good， and there was no tissue loss. The excellent rate of IHC staining and the positive rate of IHC staining in the environmental fast group were lower than those in the traditional group， but the difference was not statistically significant （χ12 = 2.769，P1 = 0.092；χ22 = 0.800，P2 = 0.375）. ③The background and outline of the two groups of WDL tissue sections were clear， the staining was uniform， the cells were clear and visible， the nuclear boundary was clear， the hybridization signal was clear and bright under the background fluorescence， and there was no miscellaneous signal. The two groups of sections were hybridized successfully， and MDM2 showed positive amplification. The number of cells successfully hybridized in the environment-friendly fast group was lower than that in the traditional group， but the difference was not statistically significant （t = 1.414，P = 0.230）. ConclusionsThe tissue treatment method of using environment-friendly sample release agent combined with ultrasound can ensure the detection effect of HE staining， IHC staining and MDM2 gene detection of pathological tissue sections， and is more efficient and environment-friendly， suitable for promotion and use in hospitals at all levels.

Interpretation and consideration of “Substitution of in vivo method(s) by in vitro method(s) for the quality control of vaccines” in General Texts of European Pharmacopoeia / 中国生物制品学杂志

Interpretation and consideration of “Substitution of in vivo method(s) by in vitro method(s) for the quality control of vaccines” in General Texts of European Pharmacopoeia@#Potency is a critical quality attribute for controlling relevant biological properties and batch consistency of vaccines.The methods can be divided into in vivo and in vitro methods according to whether animals are used.The in vivo methods are large consuming of animals and time,as well as have large variant detection results.In contrast,the in vitro alternative methods have been the hotspot of research due to their simple operations,in line with 3Rs principles,and more stable results.However,owing to the complexity of experimental design and the lack of corresponding guidance,the research progress of alternative methods is slow.Recently,“Substitution of in vivo method(s) by in vitro method(s) for the quality control of vaccines” was adopted in the European Pharmacopoeia(10th Edition),which clarifies the critical points of consideration for substitution.This paper interprets the chapter and puts forward some thoughts on that in China,which is expected to speed up the alternative methods research and improve the ability of vaccine quality control and supervision in China.

The host-targeting compound peruvoside has a broad-spectrum antiviral activity against positive-sense RNA viruses

Positive-sense RNA viruses modify intracellular calcium stores, endoplasmic reticulum and Golgi apparatus (Golgi) to generate membranous replication organelles known as viral factories. Viral factories provide a conducive and substantial enclave for essential virus replication via concentrating necessary cellular factors and viral proteins in proximity. Here, we identified the vital role of a broad-spectrum antiviral, peruvoside in limiting the formation of viral factories. Mechanistically, we revealed the pleiotropic cellular effect of Src and PLC kinase signaling via cyclin-dependent kinase 1 signaling leads to Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 (GBF1) phosphorylation and Golgi vesiculation by peruvoside treatment. The ramification of GBF1 phosphorylation fosters GBF1 deprivation consequentially activating downstream antiviral signaling by dampening viral factories formation. Further investigation showed signaling of ERK1/2 pathway via cyclin-dependent kinase 1 activation leading to GBF1 phosphorylation at Threonine 1337 (T1337). We also showed 100% of protection in peruvoside-treated mouse model with a significant reduction in viral titre and without measurable cytotoxicity in serum. These findings highlight the importance of dissecting the broad-spectrum antiviral therapeutics mechanism and pave the way for consideration of peruvoside, host-directed antivirals for positive-sense RNA virus-mediated disease, in the interim where no vaccine is available.

Mechanism of the Notch1 signaling pathway regulating osteogenic factor influences lumbar disc calcification / 中国骨伤

OBJECTIVE@#To explore the mechanism of the Notch1 signaling pathway in regulating osteogenic factors and influencing lumbar disc calcification.@*METHODS@#Primary annulus fibroblasts from SD rats were isolated and subcultured in vitro. The calcification-inducing factors bone morphogenetic protein-2 (BMP-2) and basic fibroblast growth factor (b-FGF) were added to separate groups to induce calcification, which were referred to as the BMP-2 group and the b-FGF group, respectively. A control group was also set up, which was cultured in normal medium. Subsequently, cell morphology and fluorescence identification, alizarin red staining, ELISA, and quantitative real-time polymerase chain reaction (QRT-PCR) were performed to determine the effect of calcification induction. Cell grouping was performed again, including the control group, the calcification group (adding the inducer BMP-2), the calcification + LPS group(adding the inducer BMP-2 and the Notch1 pathway activator LPS), and the calcification + DAPT group (adding the inducer BMP-2 and the Notch1 pathway inhibitor DAPT). Alizarin red staining and flow cytometry were used to detect cell apoptosis, ELISA was used to detect the content of osteogenic factors, and Western blot was used to detect the expression of BMP-2, b-FGF, and Notch1 proteins.@*RESULTS@#The induction factor screening results showed that the number of mineralized nodules in fibroannulus cells in BMP-2 group and b-FGF group was significantly increased, and the increase was greater in the BMP-2 group Meanwhile, ELISA and Western blot results showed that BMP-2, b-FGF and mRNA expression levels of BMP-2, b-FGF and Notch1 in the induced group were significantly increased (P<0.01). The results of the mechanism of Notch1 signaling pathway affecting lumbar disc calcification showed that compared to calcified group, the number of fibroannulus cell mineralization nodules, apoptosis rate, BMP-2, b-FGF content, the expression levels of BMP-2, b-FGF, and Notch1 proteins were further increased significantly However, the number of mineralization nodules, apoptosis rate, BMP-2 and b-FGF levels, BMP-2, b-FGF and Notch1 protein expression levels were decreased in the calcified +DAPT group (P<0.05 or P<0.01).@*CONCLUSION@#Notch1 signaling pathway promotes lumbar disc calcification through positive regulation of osteogenic factors.

Treatment of chronic urticaria with acupuncture and moxibustion by "mind-body regulation" based on "psychosomatic medicine" / 中国针灸

The treatment idea for chronic urticaria with acupuncture and moxibustion by "mind-body regulation" is explored in reference with "psychosomatic medicine". Chronic urticaria refers to "mind and body comorbidity" because qi and blood dysfunction and emotional abnormality are presented simultaneously in the disease and affected each other. In clinical diagnosis and treatment, the mutual regulation of mind and body should be considered to improve the curative effect. In treatment with acupuncture and moxibustion, through regulating the mind of brain, heart and zang organs, as well as qi movement, the mental activity is adjusted; through harmonizing the defensive qi and the nutrient blood, eliminating wind and stopping itching, the unhealthy conditions of body are treated. The comprehensive therapy of acupuncture is adopted in combination with moxibustion, bloodletting and auricular point therapy, in which, "regulating the mind" goes through the whole process of treatment, and the doctors and patients are well cooperated to ensure the qi-blood harmonization and the mind-body wellness.

Bioorthogonal click-to-release reaction for regulating iodo-BODIPY PDT probe / 药学学报

In this study, we synthesized six tetrazine-dipyrromethene boron difluoride (BODIPY) probes and achieved a remarkable up to 14-fold increase in singlet oxygen yield via tetrazine bioorthogonal click-to-release reactions. We systematically investigated the photodynamic activity of these probes, revealing crucial structure-activity relationships. Additionally, we evaluated the stability and release kinetics of these probes and identified P5 and P6 as ideal candidates for photodynamic therapy in live cells. This innovative strategy opens new avenues for fine-tuning the photodynamic properties of BODIPY dyes, thereby expanding their utility in cancer therapy.

Application of preoperative three-dimensional reconstruction planning in total hip arthroplasty for development dysplasia of the hip secondary to osteoarthritis / 中华骨科杂志

Objective:To investigate the clinical efficacy of preoperative three-dimensional (3D) reconstruction planning in total hip arthroplasty for development dysplasia of the hip secondary to osteoarthritis.Methods:A total of 80 patients with osteoarthritis secondary to Crowe I-III developmental dysplasia of the hip who underwent primary unilateral total hip arthroplasty from October 2019 to March 2021 were retrospectively analyzed, including 18 males and 62 females and the mean age was 55.7±10.4 years (range 41-72 years). Forty patients in the 3D group, the prosthesis type and installation angle were planed on the 3D reconstruction software based on the full-length CT scan data of the lower limbs, and the length difference of the lower limbs and hip offset were calculated. Forty patients in the control group underwent preoperative planning using conventional film measurement, and lower limb length was judged based on the preoperative measurement data and intraoperative comparison of both lower limbs. The difference of postoperative leg length, hip offset, hip function score, operating time, intraoperative blood loss, and incidence of complications were compared between the two groups.Results:All 80 patients completed the surgery successfully and the follow-up time was up to 3 months after operation. The 3D group was better than the control group in operation time (70.9±7.7 min vs. 81.6±13.3 min, t=-4.91, P<0.001), the difference of postoperative lower limb length (2.78±1.31 cm vs. 5.35±2.15 cm, t=-5.74, P<0.001), and hip function score at 1 week after operation (75.67±3.35 vs. 67.35±4.21, t=12.33, P=0.002), with statistically significant differences. In the 3D group, 95% of acetabular prosthesis and 90% of femoral stem components were consistent with the planned model, while the rate were only 75% and 68% in the control group, and the difference was statistically significant (χ 2=7.51, P=0.023; χ 2=14.92, P=0.005). There were no intraoperative complications such as vascular and nerve injury, and no postoperative complications such as dislocation or periprosthetic infection in all 80 patients. Conclusion:3D preoperative planning assisted total hip arthroplasty in the treatment of Crowe I-III developmental dysplasia of the hip secondary to osteoarthritis can improve the accuracy of the operation, and has a good clinical effect on restoring the leg length and hip offset.

Development and application of emergency management information systems for medical radiation / 中国辐射卫生

High-energy radiation derived from X-ray, γ-ray, neutrons, and other radioisotopes has been widely used for disease diagnosis and treatmentin clinical practice. Notably, high-energy radiation has been proven to increase the cure rate, prolong the survival time, and improve the quality of life among patients with malignant tumors. However, radiation poses huge threats to human health and life. Establishment of effective emergency management information systems for medical radiation is therefore of great significance to evaluate the radiation safety, predict the leakage of radioactive materials, and propose effective responses. This review summarizes the development and application of currently main emergency management information systems for medical radiation, so as to provide a reference for the establishment of sensitive and effective hospital-based radiation emergency management information systems.

Clinical efficacy of ultrasound-guided closed reduction for pediatric distal radius fracture / 中华实用儿科临床杂志

Objective:To compare the clinical efficacy of ultrasound-guided closed reduction and bare-handed reduction on pediatric distal radius fractures.Methods:Clinical data of 118 consecutive pediatric patients with distal radius fracture treated in the Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science & Technology from April 2018 to August 2019 were retrospectively analyzed.Patients treated with bare-handed reduction and ultrasound-guided closed reduction were respectively classified into group Ⅰ (58 cases) and group Ⅱ (60 cases). Baseline characteristics, treatment duration, out-patient treatment cost, postoperative pain and the modified Mayo wrist function score during the follow-up visits between groups were compared by the t test.The success rate of initial reduction, reduction times, hospitalization rate and complication between 2 groups were compared by the Chi- square test.The number of postoperative imaging scans between 2 groups was compared by the Mann- Whitney U test. Results:There were no significant differences in out-patient treatment cost and hospitalization rate between 2 groups (all P>0.05). There were significant differences in the length of stay [ (166.2±54.8) min vs.(142.6±49.2) min], success rate of initial reductions [72.4%(42/58 cases ) vs.88.3%(53/60 cases)], incidence of repeated reductions (27.6% vs.1.7%) and the incidence of postoperative repeated imaging scans (20.7% vs.1.7%) between groupⅠ and group Ⅱ (all P<0.05). A total of 104 patients (88.1%) were successfully treated with closed reduction and followed up, with a rate of success reductions.Among them, success rate of closed reduction in group Ⅰ and Ⅱ were 86.2% and 90.0%, respectively.The mean time of fracture healing was 43 (34-56) days.There were no significant differences in fracture healing time and pain score between 2 groups(all P>0.05). The modified Mayo wrist score was significantly lower in group Ⅰ than that of group Ⅱ[(97.3±4.1) points vs.(98.8±2.9) points, P<0.05]. Seven patients in group Ⅰ suffered fracture re-displacement, including 6 cases within 1 case week and 1 within 2 weeks, and 4 cases admitted to the surgical ward for further management.Five patients in group Ⅱ had re-displacement within 1 week of plaster fixation, of which 2 cases admitted to the surgical ward for further management, which was comparable between groups( P>0.05). Conclusions:Both ultrasound-guided closed reduction and bare-handed reduction are effective on the treatment of pediatric distal radius fractures, showing low incidence of complications and satisfactory functional results.Owing to its higher success rate of initial reduction, lower frequency of postoperative ra-diography and shorter length of stay, ultrasound-guided closed reduction should be recommended.

Effects of methacrylic anhydride gelatin hydrogel loaded with silver and recombinant human basic fibroblast growth factor on deep partial-thickness burn wounds in rabbits / 中华烧伤杂志

Objective: To investigate the effects of methacrylic anhydride gelatin (GelMA) hydrogel loaded with silver and recombinant human basic fibroblast growth factor (rh-bFGF) on deep partial-thickness burn wounds in rabbits. Methods: The experimental research method was adopted. Low-concentration GelMA materials, medium-concentration GelMA materials and high-concentration GelMA materials containing different concentrations of methacrylic anhydride (MA) were prepared, after adding photoinitiator, low-concentration GelMA hydrogels, medium-concentration GelMA hydrogels, and high-concentration GelMA hydrogels were obtained, respectively. The nuclear magnetic resonance spectroscopy was performed to detect the hydrogen nuclear magnetic resonance spectra of the above-mentioned three concentrations of GelMA materials, and to calculate the degree of substitution according to the spectrum diagram. The three-dimensional microstructure and pore size of 3 types of above-mentioned GelMA hydrogels were detected by field emission scanning electron microscopy (FESEM), with 9 samples measured. According to the selected concentration of MA, ten kinds of solutions of GelMA with different concentration of silver (silver-containing GelMA) were synthesized, and the silver-containing GelMA solution of each concentration was divided into three parts, and then exposed to ultraviolet light lasting for 20, 25, and 35 s, respectively. After adding photoinitiator,the corresponding silver-containing GelMA hydrogels were obtained. The residual degradation rate of silver-containing GelMA hydrogel with different photocrosslinking times was detected by collagenase degradation method at degradation of 12, 24, 36, and 48 h; and the time required for complete degradation was detected, and the sample number was 5. The inhibition zone diameter of GelMA hydrogel under above screened photocrosslinking times containing 10 concentrations of silver against Staphylococcus aureus was measured to reflect its antibacterial ability, and the sample numbers were all 5. The silver-containing GelMA hydrogel with statistical significance compared with the antibacterial circle diameter of the silver-containing GelMA hydrogel containing the lowest concentration (no silver) was considered as having antibacterial activity. The three-dimensional microstructure and pore size of the silver-containing GelMA hydrogels with antibacterial activity and the lowest drug concentration selected were detected by FESEM, and the sample numbers were all 9. The freeze-dried alone GelMA hydrogel and the freeze-dried silver-containing GelMA hydrogel were soaked in phosphate buffer solution for 24 h, respectively, then the swelling rate of the two GelMA hydrogel were calculated and compared by weighing method, and the sample number was 5. GelMA hydrogel containing silver and rh-bFGF, namely compound hydrogel for short, was prepared according to the preliminary experiment and the above experimental results. The appearance of the composite hydrogel was observed in general, and its three-dimensional microstructure and pore size were detected by FESEM. The deep partial-thickness burn wound was made on the back of 30 rabbits (aged 4-6 months, female half and half). Meanwhile, with the rabbit head as the benchmark, the wounds on the left side of the spine were treated as composite hydrogel treatment group, and the wounds on the right side were treated as gauze control group, and which were treated accordingly. On post injury day (PID) 3, 7, 14, 21, and 28, the healing of wounds in the two groups was observed. On PID 7, 14, 21, and 28, the wound healing area was recorded and the healing rate was calculated, with a sample number of 30. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and independent sample t test. Results: The substitution degree among low-concentration GelMA materials, medium-concentration GelMA materials, and high-concentration GelMA materials was significantly different (F=1 628.00, P<0.01). The low-concentration GelMA hydrogel had a loose and irregular three-dimensional spatial network structure with a pore size of (60±17) μm; the medium-concentration GelMA hydrogel had a relatively uniform three-dimensional spatial network and pore size with a pore size of (45±13) μm; the high-concentration GelMA hydrogel had the dense and disordered three-dimensional spatial network with a pore size of (25±15) μm, the pore sizes of 3 types of GelMA hydrogels were significantly differences (F=12.20, P<0.01), and medium concentration of MA was selected for the concentration of subsequent materials. The degradability of silver-containing GelMA hydrogels with different concentrations of the same photocrosslinking time was basically same. The degradation residual rates of silver-containing GelMA hydrogels with 20, 25, and 35 s crosslinking time at 12 h were (74.2±1.7)%, (85.3±0.9)%, and (93.2±1.2)%, respectively; the residual rates of degradation at 24 h were (58.3±2.1)%, (65.2±1.8)%, and (81.4±2.6)%, respectively; the residual rates of degradation at 36 h were (22.4±1.9)%, (45.2±1.7)%, and (68.1±1.4)%, respectively; the residual rates of degradation at 48 h were (8.2±1.7)%, (32.4±1.3)%, and (54.3±2.2)%, respectively, and 20, 25, and 30 s photocrosslinking time required for complete degradation of silver-containing GelMA hydrogels were (50.2±2.4), (62.4±1.4), and (72.2±3.2) h, and the difference was statistically significant (F=182.40, P<0.01), 25 s were selected as the subsequent photocrosslinking time. The antibacterial diameters of 10 types of silver-containing GelMA hydrogels against Staphylococcus aureus from low to high concentrations were (2.6±0.4), (2.5±0.4), (3.2±0.4), (12.1±0.7), (14.8±0.7), (15.1±0.5), (16.2±0.6), (16.7±0.5), (16.7±0.4), and (16.7±0.6) mm, respectively, and which basically showed a concentration-dependent increasing trend, and the overall difference was statistically significant (F=428.70, P<0.01). Compared with the silver-containing GelMA hydrogel with the lowest concentration, the antibacterial circle diameters of other silver-containing GelMA hydrogels with antibacterial ability from low to high concentration were significantly increased (with t values of 26.35, 33.84, 43.65, 42.17, 49.24, 55.74, and 43.72, respectively, P<0.01). The silver-containing GelMA hydrogel with the antibacterial diameter of (12.1±0.7) mm had the lowest antibacterial activity against Staphylococcus aureus and the lowest drug loading concentration, and the concentration of silver was selected for the concentration of subsequent materials. The microscopic morphology of the silver-containing GelMA hydrogel containing silver element with a pore size of (45±13) μm had a regular and linear strip-like structure. After soaking for 24 h, the swelling ratio of silver-containing GelMA hydrogel was similar to that of alone GelMA hydrogel. The composite hydrogel was colorless, clear and transparent, and its three-dimensional microstructure was a regular and uniform grid, with a filament network structure inside, and the pore size of (40±21) μm. On PID 3, a large amount of necrotic tissue and exudate of rabbit wound in composite hydrogel group were observed, and scattered scabs, a small amount of necrotic tissue and exudate of rabbit wound in gauze control group were observed. On PID 7, the area of rabbit wound in composite hydrogel group was significantly reduced, and adhesion of rabbit wound and gauze in gauze control group was observed. On PID 14, In composite hydrogel group, the rabbit wound surface was ruddy, and the growth of granulation tissue was observed, and in gauze control group, the rabbit wound base was pale, and the blood supply was poor. On PID 21, the rabbit wounds in composite hydrogel group healed completely, and rabbit wound in gauze control group had healing trend. On PID 28, new hair could be seen on rabbit wound surface in composite hydrogel group; oval wound of rabbit in gauze control group still remained. On PID 7, 14, 21, and 28, the wound healing areas of rabbit in composite hydrogel group were significantly larger than those in gauze control group (with t values of 2.24, 4.43, 7.67, and 7.69, respectively, P<0.05 or P<0.01). Conclusions: The medium-concentration GelMA hydrogel has good physical and chemical properties in terms of swelling and degradability. The screened silver-containing GelMA hydrogels had the lowest antibacterial activity and the lowest drug loading concentration. Composite hydrogel can significantly shorten the healing time of deep partial-thickness burn wounds in rabbits.

Formulation regularity of traditional Chinese medicine in treatment of sinusitis based on data mining / 中国中药杂志

Based on the Drugdataexpy and the prescription modern application database, this study explored the formulation regularity of ancient and modern prescriptions for the treatment of sinusitis. The Chinese medicinal prescriptions for the treatment of sinusitis with various syndromes were retrieved from the above databases and the corresponding formulation regularity was investigated by frequency analysis, association rule analysis, and factor analysis. Eighty-seven Chinese medicinal prescriptions were included, involving five syndrome types of sinusitis and 160 Chinese medicine, which were mainly effective in releasing exterior, clearing heat, and tonifying deficiency, and acted on the lung meridian due to cold and warm nature and pungent and bitter flavor or on the spleen meridian due to warm nature and pungent flavor. Seventeen core Chinese medicine were screened out by topological data analysis, including Angelicae Dahuricae Radix, Magnoliae Flos, Glycyrrhizae Radix et Rhizoma, Xanthii Fructus, and Scutellariae Radix. Chinese medicine such as Magnoliae Flos, Angelicae Dahuricae Radix, and Xanthii Fructus were commonly used in the treatment of sinusitis of wind-heat in the lung meridian, while the combination of Glycyrrhizae Radix et Rhizoma, Magnoliae Flos, Angelicae Dahuricae Radix, Chuanxiong Rhizoma, etc. was the key compatibility in treating sinusitis of dampness-heat in the spleen and stomach. Six common factors were extracted from the factor analysis of the above two syndrome types. The findings indicate that the exterior-releasing, heat-clearing, and deficiency-tonifying Chinese medicine with cold and warm nature and pungent flavor are preferential options for the clinical treatment of sinusitis. Treatment should be based on syndrome differentiation and key therapeutic principles should be followed.

Characteristics of gut microbiota in children with sepsis and influence of probiotic supplementation / 中华微生物学和免疫学杂志

Objective:To investigate the characteristics of gut microbiota in children with sepsis and the effects of probiotic intervention.Methods:Thirty-four children with sepsis admitted to the Pediatric Intensive Care Unit of Henan Provincial People′s Hospital were enrolled in this prospective study from May 2019 to July 2020. They were randomly divided into two groups and received conventional treatment (conventional treatment group, n=17) and conventional treatment combined with probiotics (probiotics group, n=17), respectively. Twenty healthy children were selected as healthy control group. The baseline characteristics and sequential organ failure assessment (SOFA) score of all children with sepsis were recorded within 24 h after recruitment. Stool samples were collected 5-7 d after recruitment. High-throughput 16S rRNA gene sequencing was used to detect gut microbiota. Bioinformatic analysis and predictive functional profiling of microbial communities were performed to analyze the differences between groups. Results:The α-diversity and β-diversity indexes showed compared with the healthy control group, the two sepsis groups had lower abundance of gut microbiota, but greater individual differences in bacteria structure. These indexes were improved significantly following probiotic intervention ( P<0.05). At the level of phylum, the proportions of Bacteroidota and Actinobacteriota in the conventional treatment group were the lowest among the three groups, while the proportion of Proteobacteria increased significantly ( P<0.05). At the level of genus, Enterocoddus was the predominant bacterium in the conventional treatment group, while the abundance of Bifidobacterium, Faecalibacterium, Erysipelotrichaceae and Rumimococcus- torques in the probiotics group showed an upward trend ( P<0.05). Differences in the abundance of metabolic pathways, including mitochondrial synthesis, exosomes, mRNA transcription and degradation and cysteine metabolism, could be found between the two sepsis groups. Conclusions:This study revealed that children with sepsis exhibit a dysbiotic microbial community with reduced microbial diversity, declined structural stability, decreased abundance of Bacteroidota and enrichment of Proteobacteria. Probiotic supplementation could elevate the percentage of beneficial symbiotic bacteria and reduce the number of pathogenic bacteria. The differential metabolic pathways might be associated with the mechanism of probiotics in practice.

Paeoniae Rubra Radix decreases the hepatotoxicity of Psoraleae Fructus in an immunologically stressed rat model: a metabolic network analysis / 药学学报

An immunologically stressed rat model was used in a metabolomics study on the ability of Paeoniae Rubra Radix to reduce the liver toxicity of Psoraleae Fructus. Different groups of rats were given the extracts of Psoraleae Fructus and Psoraleae Fructus together with Paeoniae Rubra Radix or combined with a non-toxic dose of lipopolysaccharide (LPS). The biochemical indices of liver function and pathological changes in liver tissue were used to evaluate histopathological changes. UHPLC-QTOF/MS was used to analyze the metabolic profile of serum samples, combined with principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) methods. The HMDB database and Metabo Analyst online tool were used for biomarker identification and metabolic pathway-enrichment analysis. The results show that the co-treatment Psoraleae Fructus and LPS resulted in significant liver injury, indicated by the elevation of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, as well as obvious pathological changes. Liver injury was significantly decreased by treatment with Paeoniae Rubra Radix. Metabolomic analysis showed that the addition of Paeoniae Rubra Radix ameliorated the abnormal serum metabolism in rats mainly through regulation of arachidonic acid metabolism and glycerophospholipid metabolism pathways.

Design and synthesis of tetrazine bioorthogonal fluorogenic probes / 药学学报

Bioorthogonal fluorogenic probes are becoming an ideal tool for live-cell fluorescence imaging. With the tetrazine bioorthogonal fluorogenic probe that displays fluorescence enhancement, the tetrazine plays the dual-role of a bioorthogonal reaction unit and the fluorescence quenching unit. The "off" and "on" states of the fluorescence probe are mainly controlled through inverse electron demand Diels-Alder (IEDDA) bioorthogonal reaction. We designed a series of turn-on tetrazine fluorescent probes with Donor-π-Acceptor (D-π-A) structure to achieve a high signal-to-noise ratio and specificity of fluorescence imaging. This series of probes reacted with the dienophile bicyclononyne, and then generated pyridazine structure in-situ that acted as an electron acceptor, resulting in a new D-π-A effect of fluorescent dyes, turning on the intramolecular charge transfer (ICT) effect. By adjusting the electron-donating groups and the degree of conjugation, tunable fluorescence spectra between 400-647 nm with fluorescence turn-on enhanced up to 500-fold have been achieved. This research lays the foundation for the further optimization of tetrazine bioorthogonal fluorescent probes and their applications in molecular imaging and biomedical fields.

Chromosomal microarray analysis in prenatal diagnosis of women with isolated adverse pregnancy history / 中华围产医学杂志

Objective:To investigate the abnormal results of chromosomal microarray analysis (CMA) in the subsequent pregnancy of women with adverse pregnancy history, and explore the applicability of CMA in women with different genetic etiology.Methods:Out of 5 563 pregnant women who received CMA test in Nanjing Drum Tower Hospital during June 2014 and July 2020, 169 cases that underwent prenatal diagnosis due to isolated adverse pregnancy history were retrospectively collected in this study. All the participants were divided into three groups based on the etiology type of probands, genetic origin and expected CMA outcome: high-risk group ( n=19, including 11 cases with inherited pathogenic copy number variations and eight cases with inherited chromosomal abnormalities), low-risk group ( n=113, including six cases with negative whole exome sequencing and/or CMA findings, 31 cases with confirmed monogenic disease, 47 cases with de novo pathogenic copy number variations and 29 cases with de novo chromosomal abnormalities), and unknown risk group ( n=40, none of the cases underwent genetic testing). Descriptive statistical analysis was used to summarize the abnormal detection of each group. Results:There were 169 mothers with 172 fetuses finally enrolled, including two twins and one woman with two singleton pregnancies. A total of nine cases of abnormal fetuses were detected by CMA, accounting for 5.2% (9/172). Among them, eight were in the high-risk group, which were all caused by parental abnormalities, and one case in the low-risk group was detected with a de novo 22q11.22q11.23 microduplication, which was arr[GRCh37]22q11.22q11.23(22,997,928-25,002,659)×3. No abnormality was detected in the 40 patients of unknown risk group. Conclusions:Clarifying the etiology of isolated adverse pregnancy history is crucial to the rational application of CMA. Monogenic disease, unknown cause or negative finding of CMA in probands may not be an indication for prenatal diagnosis of CMA.

Clinical efficacy of posterior percutaneous endoscopic cervical discectomy for single level cervical spondylopathy with intraspinal ossification / 中国骨伤

OBJECTIVE@#To evaluate the effectiveness and security of posterior percutaneous endoscopic cervical discectomy (PPECD) in the treatment of single level cervical spondylopathy with intraspinal ossification.@*METHODS@#Twenty three patients with single level cervical spondylopathy with intraspinal ossification were treated by posterior percutaneous endoscopic cervical discectomy between August 2017 and July 2019. There were 16 males and 7 females, aged from 29 to 74 years old with an average of (50±13) years.The disease duration were 3 to 120 months with a median of 6 months. There were 9 cases of cervical spondylotic radiculopathy, 6 cases of cervical spondylotic myelopathy, and 8 cases of mixed cervical spondylopathy. According to the characteristics of ossification, 17 cases were osteophytes on the posterior edge of the vertebral body;3 cases were protrusion ossification;3 cases were posterior longitudinal ligament ossification. According to the position of ossification in spinal canal, 14 cases were medial and lateral type, 5 cases were central type, and 4 cases were mixed type. Posterior percutaneous cervical endoscopic cervical discectomy in patients performed by the same surgeon. Japanese Orthopaedic Association (JOA) score and visual analogue scale(VAS) were compared separately before and after operation. At 3 months after operation, clinical effect was assessed according to modified Macnab standard.@*RESULTS@#All operations were successful. The operative time was 30 to 155 (69.1±27.2) min. The bedridden time was 2 to 3(3.0±0.9) h, length of postoperative hospitalization was 2 to 7(4.1± 1.5) d. Three dimensional CT reconstruction of the cervical spine at 3 days after operation showed that ossified tissue of 13 cases were completely removed, and 10 cases were left after operation, and the residual was located at the posterior edge and/or center of the upper vertebral body. VAS score at discharge from hospital was significantly lower than that before operation (@*CONCLUSION@#For an experienced surgeon, percutaneous posterior cervical endoscopic discectomy is safe and reliable in treating single level cervical spondylopathy with intraspinal ossification, and can obtain good clinical results.

Research advances in mechanism of gut microbiota changes after traumatic brain injury / 中华创伤杂志

Traumatic brain injury (TBI) is often accompanied by digestive system dysfunction, such as intestinal dysfunction and dysbacteriosis. The activation of neuroendocrine system after TBI is one of the factors leading to gut microbiota changes. A variety of antimicrobial peptides secreted by Paneth cells are the key factors to maintain intestinal flora homeostasis. The dysfunction of Paneth cells after TBI may be responsible for gut microbiota changes. The authors review the mechanism of gut microbiota changes induced by TBI to provide new ideas for clinical treatment of gut microbiota changes caused by TBI.

Expert consensus on rehabilitation strategies for traumatic spinal cord injury / 中华创伤杂志

TSCI have dyskinesia and sensory disturbance that can cause various life-threaten complications. The patients with traumatic spinal cord injury (TSCI), seriously affecting the quality of life of patients. Based on the epidemiology of TSCI and domestic and foreign literatures as well as expert investigations, this expert consensus reviews the definition, injury classification, rehabilitation assessment, rehabilitation strategies and rehabilitation measures of TSCI so as to provide early standardized rehabilitation treatment methods for TSCI.

Analysis of curative effect of Doppler ultrasound-guided management for vascular complications of Gartland type Ⅲ supracondylar fractures of the humerus in children / 中华实用儿科临床杂志

Objective:To investigate the curative effect of Doppler ultrasound-guided management of vascular injury of Gartland type Ⅲ supracondylar fractures of humerus in children.Methods:A prospective study on 18 children with vascular complications of pulseless Gartland type Ⅲ supracondylar humeral fracture who were admitted at Department of Pediatric Orthopedic Surgery, Wuhan Children′s Hospital from March 2017 to March 2018 was conducted.Among these children, 12 were male and 6 were female, with the age of 2.1-8.6 years (mean 4.4 years old), and 10 cases were injured on the left and 8 cases on the right.All patients were satisfied with closed reduction and internal fixation within 24 hours after injury.Before the operation, Doppler ultrasound was performed to determine the shape of brachial artery and it relationship with fracture.During reduction, Doppler ultrasound was used to assess brachial pulse and blood perfusion.Then, the brachial artery was assessed by palpable radial pulse and peripheral blood supply, and elbow joint function was evaluated with Mayo Elbow Performance Score and Flynn criteria.Results:Eighteen patients were followed up for 6 to 12 months (average 9 months) after operation.Before reduction, radial pulse disappeared and peripheral blood supply was good in 18 cases.Preoperative Doppler ultrasonography showed that the brachial artery was located in front of the proximal humerus fracture fragment.A proximal stream of the brachial artery was identified, but the distal blood vessels were compressed by the fragment.A stream of radial artery was identified in 5 patients with Doppler, and no stream was identified in 13 patients.After reduction, the peripheral blood supply of all the affected limbs was good, among which the peripheral blood supply, 1 case returned to normal after 5 minutes waiting.Specifically the palpable radial pulse was immediately restored in 14 patients.The palpable radial pulse did not restore but the peripheral blood supply was good in 4 cases.Intraoperative ultrasonography showed that brachial artery blood flow was good, and palpable radial pulse was restored between 3 to 5 weeks later.In patients with at average sixteen-week neurological follow-up, 5 patients had complete resolution of nerve palsy.No complications such as forearm compartment syndrome and elbow joint dysfunction occurred.Functional outcome as measured by Flynn criteria was excellent in 12 patients, good in 6 patients, the excellent and good rate was 100%.Functional outcome as measured by Mayo Elbow Performance Score was excellent in 17 patients, good in one patient, the excellent and good rate was 100%.Conclusion:Ultrasound-guided is a safe and reliable option to treat pulseless Gartland type Ⅲ supracondylar humeral fracture.

Analysis of correlation between clinical phenotypes and genotypes in children with distal renal tubular acidosis / 中华实用儿科临床杂志

Objective:To analyze the correlation between clinical phenotypes and genotypes in 6 children with primary distal renal tubular acidosis (dRTA).Methods:The clinical data of 6 children confirmed as dRTA in Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science & Technology from November 2017 to August 2019 were collected, and related auxiliary examination was performed to assess their growth and development.The venous whole blood was reserved for Trio whole exome sequencing, and full spectrum genetic disease accurate diagnosis cloud platform was applied to systematic data screening and analysis.The suspected mutations were checked by Sanger sequencing, and then the role of protein was predicted by software.Results:Clinical manifestations, signs and auxiliary examination results of the 6 children accorded with the diagnostic criteria of dRTA, and the prominent characteristics was growth retardation.One case had knee valgus, one had osteoporosis, and the auxiliary examination results showed that both of them had alkaline urine, metabolic acidosis, and hypokalemia.Three children had nephrocalcinosis, and 2 children had nephrolithiasis.The parents of the 6 patients were all normal without phenotypes.Mutations in the SLC4A1 gene were identified in 4 patients, including 1 child with a reported homozygous autosomal recessive missense mutation(c.2102G>A, p.G701D), who had dRTA and hemolytic anemia, and 3 children with the reported de novo heterozygous autosomal dominant missense mutation(c.1766G>A, p.R589H, c.1765C>T, p.R589C), whose age at diagnosis was related to abnormal renal imaging.Compound heterozygous autosomal recessive mutations in the ATPV1B1 gene were identified in 1 patient, and they were novel heterozygous missense mutations (1153C>A, p.P385T and c. 806C>T, p.P269L). A novel homozygous autosomal recessive missense mutation was identified in 1 patient in the ATPV0A4 gene(c.1899C>A, p.Y633X, 208). Conclusions:Mutations in SLC4A1, ATP6V1B1, ATP6V0A4 genes are identified as the main causes of the primary dRTA, and the phenotypes was related to the mutation features and genotypes.Genetic test should be conducted on patients suspected as dRTA for early molecular diagnosis, thereby improving clinical phenotypic screening and individualized treatment.