Objective To investigate the effect of total flavones of Rhizoma Drynariae on femur distraction osteogenesis in the rabbits. Methods Thirty-two healthy rabbits were randomly divided into treatment group and control group, 16 rabbits in each group. The femoral fracture was treated with unilateral femoral distraction and was fixed with a self-made distraction instrument. After 7-day intermittent period, the fractured femur was distracted at a rate of 1 mm/d, twice a day for 10 continuous days. The treatment group was fed with total flavones of Rhizoma Drynariae from the first post-operative day to the end of the experiment. And then all of the animals were sacrificed after fixation for 28 days. The bone specimens were used for histological observation and immunohistochemical detection. Results The area of mature bone in the newborn bone tissue of the treatment group was increased, and osteoblasts number and the percentage of trabecular bone area were significantly higher than those of the control group . The bone morphogenetic protein-2 (BMP-2) and transforming growth factor-β1 (TGF-β1) were stained brown deeply, the staining degree being stronger than that of the control group. Conclusion Rhizoma Drynariae total flavones can effectively accelerate the formation and maturation of newborn bone tissue during bone distraction.

Objective To investigate the clinical efficacy of Jin’s three-needle acupuncture plus intelligence training in treating mental retardation in children under 6 years old. Methods Three hundred mental retardation patients aged 0 to 6 were randomly allocated, according to their parents’ willingness, to treatment and control groups, 150 cases each. The treatment group received Jin’s three-needle acupuncture plus intelligence training and the control group, intelligence training alone. An assessment was made using Chinese revised version of the Gesell Developmental Schedules in both groups before and after treatment.Results The total efficacy rate was 84.7% in the treatment group and 73.3% in the control group; there was a statistically significant difference between the two groups (P<0.05). The total efficacy rates in liver-kidney deficiency, heart-spleen deficiency, phlegm and blood stasis, and qi deficiency and blood stasis types were 86.8%, 86.8%, 76.5% and 75.0%, respectively, in the treatment group and 76.2%, 76.8%, 62.5% and 60.0%, respectively, in the control group. There were statistically significant differences in the total efficacy rates in different syndrome types between the two groups (P<0.05). The total efficacy rates in the patients with mild, moderate, severe and very severe diseases were 96.9%, 94.7%, 66.7% and 44.4%, respectively, in the treatment group and 90.0%, 82.5%, 47.8% and 35.3%, respectively, in the control group. There was a statistically significant difference in the total efficacy rate between the two groups of patients with moderate, severe or very severe disease (P<0.05).Conclusion Jin’s three-needle acupuncture plus intelligence training is an effective way to treat mental retardation in children.

BACKGROUND: Ankle fracture combined with deltoid ligament rupture and distal tibiofibular syndesmosis injury occurs occasionally. Its treatment with distal tibiofibular syndesmosis screw fixation or deltoid ligament repair remains controversial. The former appears with poor reduction, broken nails, secondary surgery and other problems.OBJECTIVE: To observe the clinical effectiveness of suture anchor repair for ankle fracture combined with deltoid ligament injury.METHODS: Twelve patients with ankle fracture combined with deltoid ligament injury were selected from the First Affiliated Hospital of Guangzhou University of Chinese Medicine between January 2013 and December 2015. All patients were treated with open reduction, internal fixation, and anchor repair, but without distal tibiofibular syndesmosis screw fixation. The curative efficacy and joint stability were observed.RESULTS AND CONCLUSION: (1) All patients were followed-up for more than 12 months. (2) The modified Baird-Jackson scoring system showed excellent in nine cases, good in two cases, average in one case, poor in none case, and the excellent and good rate was 92%. (3) At 1 year after internal fixation, the X-ray examination showed the malleolus gap and talus slope angle of the affect side were (3.47±0.12) mm and (0.45±0.18)°, and the malleolus gap and talus slope angle of the healthy side were (3.44±0.05) mm and (0.43±0.14)°, and there was no significant difference between two sides (P > 0.05). (4) These results indicate that the suture anchor can repair the anatomy and biomechanics of deltoid ligament with stable ankle joint, and secondary surgery is unnecessary.

Objective To evaluate the application of real-time three-dimensional transesophageal echocardiography (RT-3D TEE)in left atrial appendage (LAA)closure.Methods After excluding valvular disease,21 of 36 atrial fibrillation (AF)patients suffered the percutaneous LAA closure were chosen to measure left ventricular ejection fraction (LVEF)and to observe if the thrombus or the cloudiness echo will occur.The multiple planner reconstruction function was applied to rebuild sections and to observe the LAA anatomical morphology and its internal structure.During operation,the Flexi Slice function was used to measure the dimensions of LAA ostium,RT-3D mode was used to monitor sheath transport and closure device release,and to immediately evaluate the effect of blocking and complications after the operation. Results The exam before the closure procedure showed 5 of 36 patients' left ventricular ejection fraction (LVEF)<40% and one or more thrombus or the cloudiness echo were found in other 10 patients' which were thus excluded from the study.RT-3D TEE examination in the 21 patients revealed 8 single-lobe cases, 8 double-lobes cases and 5 patients with multi-lobe.Two patients of them need a special device.The maximum dimensions of LAA ostium was (22.24±4.35)mm,the fixed plate size of conventional LAmbreTM device was (28.26±5.23)mm.All patients choiced an average (1 .1 ±0.30)closure device and get the best sealing effect evaluated by RT-3D TEE immediately after operation,10 cases of them has about 1 -3 mm residual leakage exist around the closure.None of the patients with a new pericardial effusion and cardiac tapenade,no closure shift and embolism events.Conclusions RT-3D TEE can real-time and dynamically observe LAA anatomical morphology and its internal structure,compared with other imaging techniques, RT-3D TEE has irreplaceable advantages in the percutaneous LAA closure.

Objective To observe clinical efficacy and safety on the treatment of moderate-severe cancer pain by Fluvoxamine combined with Oxycodone. Methods 120 cancer patients with moderate pain and 120 cases with severe pain were selected, randomly divided into experimental group and control group. The control group were given oxycodone alone , and experimental group given fluvoxamine combined with oxycodone , till the pain relieved, then the degree of pain relief, oxycodone dosage, life quality and side effects were evaluated. Results The degree of pain relief in experimental group were much better than control group (P < 0.05). Oxycodone consumption were lower in experimental group than control group , and the difference was no significant difference in controlling moderate pain (P = 0.065), while statistically significant in controlling severe pain (P = 0.035). The general status, daily activity, mood, and sleep affected by cancer pain were released after treatment, especially in experimental group (P < 0.05). The most common side effects were approximative, and the incidence of constipation, nausea/vomiting, lethargy were lower in experimental group than control group (P =0.026). Conclusion Fluvoxamine combined with Oxycodone can effectively control moderate-severe cancer pain, and reduce the oxycodone dosage and some side effects, and therefore, improve the quality of life.

Objective To explore the predictive value of neutrophil-to-lymphocyte ratio on the prognosis of H7N9 avian influenza.Methods A retrospective analysis was conducted on 28 H7N9 avian influenza patients (treatment group) at the First Affiliated Hospital of Soochow University from April 2013 to January 2016.Thirty healthy physical examiners in the same period were enrolled as the healthy control group.The 28 patients were followed up for half a year and divided into the improvement group (18 cases) and the death group (10 cases) according to the clinical prognosis.Inflammatory indicators including white blood cells (WBC),neutrophil (N),lymphocyte (L),monocytes (M),platelet (PLT),creatine kinase (CK),lactate dehydrogenase (LDH),high sensitive C reactive protein were collected at day 1,day 3 and week 1 of admission.Calculation of neutrophil-to-lymphocyte ratio (NLR),platelet-to-lymphocyte ratio (PLR),lymphocyte-to-monocyte ratio (LMR),△NLR3 (day 3 of admission NLR-on day 1 of admission NLR),△NLR7 (week 1 of admission NLR-day 3 of admission NLR) and so on calculating △PLR3,△PLR7,△LMR3,△LMR7.Differences of the above indicators between the improvement group and death group were compared.The measurement data with normal distribution were tested by t-test of two independent samples,and the count data with non-normal distribution were tested by Mann-Whitney U-test.Univariate and multivariate logistic regression analysis to explore the prognostic factors and the working characteristic curve of subjects was used to evaluate the predictive value of inflammatory response indexes for H7N9 avian influenza death.Results In the treatment group,the baseline WBC,L,N,PLT,the proportion of lymphocytes,neutrophils,monocytes,and NLR,PLR,and LMR were all statistically different compared with the healthy control group (all P ＜0.01).After treatment,day 3 NLR,△NLR3 in improvement group were both significantly decreased to 10.93 (15.71)and0.87 (-15.63),respectively when compared with death group (17.62[23.63] and 7.42[22.68],respectively) (Z =-2.16 and-2.014,respectively,both P＜0.05).Day 7 NLR,△NLR7 in improved group were 6.51 (13.23) and-0.37 (-12.38),respectively,which were both lower than those of death group (27.90 [25.64] and 11.54 [-26.22]) with statistically significant differences (Z =-2.444 and -2.111,respectively,both P ＜ 0.05).Multivariate logistic regression analysis indicates that △NLR3 is the main factor that affects the prognosis of the H7N9 infection (odds ratio [OR] =1.153,95％ confidence interval [CI]:1.052-1.263,P =0.002).Reciver operating characteristic curve analysis showed that the area under the curve was 0.733 (95 ％ CI:0.532-0.935,P =0.044).Based on the principle of Youden index,the cutoff value of △NLR3 to predict the death risk of H7N9 avian influenza was 5.453 with sensitivity of 0.700 and the specificity of 0.722.The mortality was higher when △NLR3 was higher than 5.453.Conclusions Dynamic monitoring NLR,especially △NLR3 may reflect the condition and prognosis of H7N9 infection,which is an independent predictor of death.

To explore the immunomodulatory effects of interleukin-17 (IL-17) on acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods Thirty-six SPF-class C57BL/6 mice were divided into normal saline control group (NS group) and LPS-induced ALI model group (LPS group, LPS 5 mg/kg intratracheal drip) according to random number table method, with 18 mice in each group. Six mice were sacrificed at 2, 6 and 24 hours after model reproduction, and peripheral blood, lung and spleen tissues were harvested. After staining with hematoxylin-eosin (HE), the pathological changes of lung tissue were observed under microscope and the infiltration level of lymphocytes, neutrophils and macrophages in the alveolar wall and tracheal wall were detected. Immunohistochemistry was used to detect the protein expression of IL-17 in alveolar wall and tracheal wall, and the correlation between IL-17 expression and lymphocytes, neutrophils and macrophages infiltration in alveolar wall and tracheal wall were analyzed. The level of IL-17 in lung tissue homogenate was determined by enzyme linked immunosorbent assay (ELISA). Flow cytometry was used to detect the proportion of CD4+IL-17+ helper T cells (Th17 cells) in CD4+ T cells in peripheral blood, lung tissue and spleen tissue. Results ① Microscopy showed that the lung tissue structure of NS group was basically normal at each time after model reproduction, and there was no obvious inflammatory cell infiltration, while the lung tissue edema and inflammatory reaction were gradually aggravated in the LPS group, and the lung injury score was significantly higher than that in NS group at each time (2 hours: 4.47±1.42 vs. 1.10±0.55, 6 hours: 7.93±2.14 vs. 1.23±0.50, 24 hours:12.67±2.67 vs. 1.20±0.61, all P < 0.01). ② Immunohistochemistry showed that the protein expression of IL-17 in alveolar wall and tracheal wall of LPS group increased gradually with time, while that in NS group was negative or weak positive. Quantitative analysis showed that the immunohistochemical staining score of IL-17 protein in alveolar wall and tracheal wall of LPS group were higher than those of NS group (alveolar wall: 2.70±1.40 vs. 0.90±0.37 at 2 hours, 5.10±1.76 vs. 1.17±0.59 at 6 hours, 9.67±1.32 vs. 1.10±0.45 at 24 hours; tracheal wall: 2.87±0.89 vs. 0.90±0.39 at 2 hours, 4.97±1.48 vs. 1.10±0.41 at 6 hours, 8.67±1.54 vs. 1.03±0.29 at 24 hours; all P < 0.05). ③ Correlation analysis showed that the protein expression of IL-17 in alveolar wall and tracheal wall were positively correlated with the degree of lymphocyte, neutrophil and macrophage infiltration (alveolar wall: r value was 0.632, 0.550, 0.466; tracheal wall: r value was 0.695, 0.662, 0.575, respectively; all P < 0.01). ④ IL-17 content (μg/L) in lung tissue homogenate was significantly higher than that in NS group at each time after model reproduction (2 hours: 1.37±0.14 vs. 1.01±0.18, 6 hours: 1.65±0.19 vs. 1.11±0.18, 24 hours: 1.92±0.36 vs. 1.17±0.24, all P < 0.01). ⑤ The proportion of Th17 cells in the peripheral blood, lung tissue and spleen tissue of the LPS group were higher than those of the NS group at each time after model reproduction [peripheral blood: (2.62±0.62)% vs. (1.42±0.40)% at 2 hours, (3.74±0.43)% vs. (1.27±0.32)% at 6 hours, (4.44±0.65)% vs. (1.59±0.45)% at 24 hours; lung tissue: (2.32±0.44)% vs. (1.50±0.25)% at 2 hours, (3.66±0.36)% vs. (1.33±0.24)% at 6 hours, (4.60±0.54)% vs. (1.60±0.27)% at 24 hours; spleen tissue: (1.49±0.36)% vs. (0.69±0.21)% at 2 hours, (2.58±0.55)% vs. (0.59±0.18)% at 6 hours, (3.76±0.57)% vs. (0.65±0.26)% at 24 hours; all P < 0.01]. Conclusion IL-17 is involved in the inflammatory immune regulation of ALI mice.

Objective:To investigate a rapid protocol for the acquisition of a three-dimensional (3D) image of corneal nerve distribution and its various parameters.Methods:Four SPF female C57BL/6 mice were selected and four corneal samples with complete limbi were obtained using a dissecting microscope after the sacrifice of mice euthanized by ether.After conventional fixation, permeabilization, and immunostaining by an anti-β-Ⅲ tubulin fluorescent-conjugated antibody, a whole-nerve image of the whole-mount cornea was captured under a 60X oil lens using a scientific complementary metal-oxide-semiconductor detector and a high-resolution deconvolution microscope.The 3D image of the corneal nerve fiber was obtained after 3D deconvolution processing, Z-axis data projection, and automatic stitching using the self-contained image processing software of the microscope system.The corneal nerve density in different areas was analyzed using the automatic detection mode in the Filament Tracer module and the manual Autopath module of the interactive microscopic image analysis software Imaris.The use and care of animals complied with the statement of the Association for Research in Visual and Ophthalmology, and the study protocol was approved by an Experimental Animal Welfare Committee of Jinan University (No.JN-A-2002-01).Results:It was found that stromal nerve fibers in a dense network entered the Bowman membrane near the limbus, and branches of stromal nerve fibers formed subbasal nerve plexus, which stretched toward the center of the cornea to form a dense neural network-like structure and converged into a vortex-like structure at the apex of the cornea.Some subbasal nerves entered the epithelial layer vertically and some branches of nerve endings were found.Through the automatic detection mode of Imaris software, a gradual increase of the density from (2 488.88±282.84)μm/μm 2 at the limbus to (5 766.66±298.55)μm/μm 2 at the center of the cornea of the subbasal nerve branches, and a decrease of the density from (40.99±0.99)μm/μm 2 at the limbus to (34.57±1.28)μm/μm 2 at the center of the stromal nerves were found.It was also found that the stromal nerves at the limbus entered the Bowman membrane for about 151 μm and then began to branch to form subbasal nerves. Conclusions:The high-resolution deconvolution microscope system can be used to study the 3D distribution of the whole corneal nerve.Additionally, Imaris can be used for obtaining various parameters of the corneal nerves automatically and quickly.

OBJECTIVE: To establish a non-invasive method for beta-thalassemia by detecting parental CD41-42 mutation in cell-free DNA derived from maternal plasma with droplet digital PCR (ddPCR). METHODS: Beta-actin gene and beta-thalassemia gene CD41-42 mutation were respectively set as the reference and target sequences. A novel method was established based on Bio-Rad ddPCR technique with specific primers and TaqMan probes for the two genes. The accuracy, sensitivity and detective linearity range of the developed method were evaluated by detection of the target gene gradient concentration samples. The applicability was also evaluated by testing 20 maternal plasma samples. RESULTS: The ddPCR method could accurately detect the beta-thalassemia CD41-42 mutation in cell-free DNA derived from maternal plasma. Within the target sequence concentration ratio of 5.00%-0.50%, the relative errors were all < 0.05, the linear regression equation was Y=1.0101-X-0.0071 and R=0.9994. The results of 20 maternal plasma cell-free DNA samples were all consistent with those of the follow-up testing. CONCLUSION A ddPCR method for detecting parental CD41-42 mutation in cell-free DNA from maternal plasma was developed. The method is simple, rapid, accurate, and can be applied for non-invasive prenatal diagnosis for couples simultaneously carrying the CD41-42 mutation.
Cell-Free Nucleic Acids ; DNA ; blood ; Female ; Humans ; Mutation ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; methods ; beta-Thalassemia ; diagnosis ; genetics

Objective:To analyze the influencing factors of death in patients with acute respiratory distress syndrome (ARDS) combined with acute kidney injury (AKI) in intensive care unit (ICU) after continuous renal replacement therapy (CRRT).Methods:The demographic and clinical data of ICU patients with ARDS combined with AKI after CRRT at Henan Provincial People's Hospital from January 1, 2018 to December 31, 2018 were collected. According to the final treatment results of this hospitalization, the patients were divided into death group and survival group. Survival was defined as the improved patient's condition and hospital discharge. Death was defined as the patient's death during the ICU hospitalization or confirmed death after abandoning treatment and automatically being discharged from the hospital in the follow-up. The basic clinical characteristics and CRRT status between the two groups were compared. Multivariate logistic regression method was used to analyze the influencing factors of death in patients.Results:A total of 132 patients were enrolled, of which 90 patients (68.2%) died, with 84 males (63.6%) and median age of 59(45, 73) years. Compared with the survival group, the death group had higher age, proportion of malignant tumors, sequential organ failure assessment (SOFA) score, number of organ dysfunction and proportion of positive balance of fluid accumulation at 72 hours, longer time from entering ICU to CRRT, and lower mean arterial pressure (minimum value) and oxygenation index (all P<0.05). Multivariate logistic regression analysis results showed that the age≥60 years old ( OR=4.382, 95% CI 1.543-12.440, P=0.006), large number of organ dysfunction ( OR=1.863, 95% CI 1.109-3.130, P=0.019), high SOFA score ( OR=1.231, 95% CI 1.067-1.420, P=0.004) and long time from ICU admission to CRRT ( OR=1.224, 95% CI 1.033~1.451, P=0.020) were independent influencing factors of death in patients with ARDS combined with AKI after CRRT, and high oxygenation index ( OR=0.992, 95% CI 0.986-0.998, P=0.010) was an independent protective factor for patients' prognosis. Conclusions:The mortality of patients with ARDS combined with AKI after CRRT is still high. The age≥60 years old, large number of organ dysfunction, high SOFA score and long time from ICU admission to CRRT are independent influencing factors for death, and high oxygenation index is an independent protective factor for prognosis in patients with ARDS combined with AKI after CRRT.