Mechanism of Tongxie yaofang reducing colon hypermotility in IBS-D rats by regulating gut microbiota-ENS-MM crosstalk / 中国药房

OBJECTIVE To study the effects of Tongxie yaofang reducing colon hypermotility in irritable bowel syndrome with diarrhea （IBS-D） rats by regulating gut microbiota-enteric nervous system （ENS）-muscularis macrophages （MM） crosstalk. METHODS Forty newborn male SD rats were randomly divided into control group， model group， TCM group [Tongxie yaofang 2.68 g/（kg·d）， calculated by raw material]， and positive control group [Live combined bifidobacterium and lactobacillus tablets 0.27 g/（kg·d）]， with 10 rats in each group. Except for the control group， the IBS-D model of liver stagnation and spleen deficiency syndrome was induced in the other 3 groups with the method of mother-child separation+chronic restraint+Folium Sennae- induced diarrhea. After modeling， the administration groups were given relevant drug liquid intragastrically， once a day， for consecutive 2 weeks. At the end of modeling and after administration， the fecal properties （the incidence and the rate of loose stools）， colonic motility （colon emptying time）， and visceral sensitivity [abdominal withdrawal reflex （AWR） scores under different pressures] of rats were observed in each group. The concentration of lipopolysaccharide （LPS） in serum was detected after medication， and the expressions and distribution of bone morphogenetic protein 2 （BMP2）， colony stimulating factor 1 （CSF1） and Toll-like receptor 4 （TLR4） in colon tissue were detected； the diversity， species composition and differences of gut microbiota were also determined. RESULTS At the end of modeling， compared with the control group， all rats of the other three groups suffered from loose stools （100%）， the rate of loose stools and AWR scores at different pressures increased significantly， and colon emptying time was shortened significantly （P＜0.01 or P＜0.05）. The incidences of loose stools were 20% in TCM group and 80% in the positive control group； the rate of loose stools and AWR scores at different pressures， serum concentration of LPS and protein expressions of CSF1 and TLR4 in muscle layer of colon tissue in TCM group significantly decreased， compared with the model group； colon emptying time， the average optical density of BMP2 protein in muscle layer of colon tissue， and the indexes of Chao 1， Shannon and Faith’s PD and Simpson E-mail：772699670@qq.com index of rats in TCM group were all prolonged or increased significantly， compared with the model group （P＜0.01 or P＜ E-mail：aiwangzi312@163.com 0.05）. The relative abundance ratio of Firmicutes/Bacteroidetes， from low to high， was in the order of TCM group， control group， positive control group and model group； the species composition of gut microbiota in TCM group was closer to control group， with dominant bacterial genera including Prevotella and Blautia. CONCLUSIONS Tongxie yaofang can regulate the expressions of BMP2 and CSF1， the key proteins of gut microbiota-ENS-MM crosstalk， by changing the gut microbiota， thus alleviating the abnormal hyperfunction of colon motility in IBS-D rats.

Molecular mechanism of sulforaphane promoting the differentiation of bone marrow stem cells into osteoblasts / 局解手术学杂志

Objective To investigate the molecular mechanism of sulforaphane(Sul)promoting bone marrow stem cells(BMSCs)differentiating into osteoblasts.Methods BMSCs were divided into the control group(without any treatment),induction group(induction of osteogenic differentiation),and induction+Sul group(induction of osteogenic differentiation with the addition of 40 μmol/L of Sul).The adenovirus-shRNA-Mock,-shRNA-TET1,-shRNA-TET2,and-shRNA-TET3 were transfected into BMSCs as the shRNA-Mock group,shRNA-TET1 group,shRNA-TET2 group,and shRNA-TET3 group.BMSCs were cultured in cell culture medium containing osteogenic differentiation induction medium and 40 μmol/L of Sul,and then transfected with adenovirus-shRNA-TET1,-shRNA-TET2,-shRNA-TET3,and-shRNA-Mock as the induction+Sul+shRNA-TET1 group,induction+Sul+shRNA-TET2 group,induction+Sul+shRNA-TET3 group,and induction +Sul+shRNA-Mock group.The mRNA and protein expression levels of Runx2 after BMSCs differentiated into osteoblasts were determined by qPCR and Western blot.The DNA content of Runx2 promoter region bound to Histone H3 after BMSCs differentiated into osteoblasts was determined by chromatin immunocoprecipitation(ChIP).The methylation level of Runx2 promoter region of BMSCs differentiated into osteoblasts was determined by HpaⅡenzyme and MspⅠenzyme digestion combined with qPCR.The degree of BMSCs differentiated into osteoblasts was determined by alizarin red staining.Results Compared with the induction group,the mRNA and protein expression levels of Runx2 in the induction+Sul group were significantly increased(P<0.05);the content of DNA in the Runx2 promoter region bound to Histone H3 was increased(P<0.05),the methylation level of Runx2 promoter region was reduced(P<0.05),and the alizarin red staining score was elevated(P<0.05).Compared with the induction+Sul group,the content of DNA in the Runx2 promoter region bound to Histone H3 in the induction+Sul+shRNA-TET1 group was decreased(P<0.05),the methylation level of Runx2 promoter region was increased(P<0.05),and the alizarin red staining score was decreased(P<0.05).While there was no significant change among the induction+Sul+shRNA-TET2 group,induction+Sul+shRNA-TET3 group,induction+Sul+shRNA-Mock group(P>0.05).Conclusion Sul can promote the differentiation of BMSCs into osteoblasts through promoting DNA demethylation of Runx2 promoter region by TET1.

Transfer of polyfoliate perforator flap of fibular hallux in reconstruction of multiple digit-tip defects / 中华显微外科杂志

Objective:To evaluate the clinical effectiveness of the reconstruction of multiple digit-tip defects with transfer of polyfoliate perforator flaps of the fibular hallux.Methods:From January 2019 to June 2022, 15 patients had undergone reconstruction surgery for multiple digit-tip defects using polyfoliate perforator flaps of ipsilateral fibular hallux, with the first dorsal metatarsal artery as the pedicle, in the Department of Upper Limb Repair and Reconstruction Surgery, Guizhou Hospital of Beijing Jishuitan Hospital. The patients were 10 males and 5 females and aged 20 to 45 years old. Eight patients had the defects of thumbs and index fingers, 4 of thumbs and middle fingers, 2 of thumb, index and middle fingers and 1 of thumb, index and ring fingers. All the 15 digit injuries had nail bed defects to which reconstructive surgery were required. For the flaps of dorsal artery, flaps were 1.8 cm×2.0 cm-2.0 cm×3.1 cm in size and for those of plantar artery, the flaps sized 1.5 cm×2.0 cm-2.5 cm×3.0 cm. Donor site defects in the hallux were reconstructed with free superficial circumflex iliac perforator flaps. Postoperative follow-up lasted until 30th June 2023 and included visits to the outpatient clinic, WeChat and telephone reviews to assess the appearance, function and sensation recovery of the digits.Results:All the 15 flaps survived. During the 6 to 24 months (16 months in average) of postoperative follow-up, the appearance and texture of all flaps were found close to the healthy digits, with good nail growth and without deformity. TPD were found between 8.0 mm and 12.0 mm. The donor sites on the great toes that reconstructed with superficial circumflex iliac artery flaps were all survived well, and the incisions were satisfactorily healed without the functions of walking, running or jumping being significantly affected.Conclusion:The use of polyfoliate perforator flaps of fibular hallux for reconstruction of multiple digit-tip defects is an ideal surgical method due to the consistency of vascular anatomy, ease with flap harvest, similarity in the normal digital skin texture, and the capability to include a nail bed with the flap. A single donor from the hallux can simultaneously reconstruct two defects of digit-tip, making it an excellent treatment in the reconstruction of small-to medium-sized composite tissue defects in multiple digits.

Diagnostic value of ultrasound-based hemodynamic examination of superior mesenteric artery for acute suppurative appendicitis / 医疗卫生装备

Objective To investigate the characteristics of hemodynamic changes in superior mesenteric artery(SMA)of patients with acute suppurative appendicitis(ASA)to facilitate the diagnosis of ASA.Methods Ninety-three ASA patients diagnosed by ultrasound and confirmed by pathology in some hospital from January 2015 to December 2022 were enrolled into an ASA group,and 88 soldiers without abnormalities in the annual physical examination at the hospital in 2021 were divided into a control group,and the two groups were compared in terms of SMA hemodynamic indexes including peak systolic velocity(PSV),end diastolic velocity(EDV),resistance index(RI)and systolic/diastolic ratio(S/D).The statistically significant data of the above indexes were selected to draw the ROC curve and obtain the cut-off values.SPSS 22.0 software was used for statistical analysis.Results The PSV was(212±69)cm/s in the ASA group and(118±26)cm/s in the control group,with statistically significant differences(P＜0.05),and the two groups had the differences in EDV,RI,and S/D not statistically significant(P＞0.05).The ROC curve was plotted based on the PSV data,and a cutoff value of 238 cm/s was obtained with an AUC of 0.714.Conclusion Ultrasound examination of SMA hemodynamic changes in suspected ASA patients facilitates a definitive diagnosis and is of guidance for clinical treatment.[Chinese Medical Equipment Journal,2023,44(10):64-67]

Dihydromyricetin improves cardiac insufficiency by inhibiting HMGB1 in diabetic rats / 南方医科大学学报

OBJECTIVE@#To investigate the effect of dihydromyricetin (DHM) on cardiac insufficiency in diabetic rats and explore the underlying mechanism.@*METHOD@#Twenty-four male SD rats were randomized equally into normal control group, type 2 diabetes (T2DM) group fed on a high-glucose and high-fat diet for 6 weeks with low-dose streptozotocin (STZ) injection, metformin (MET) group with daily intragastric administration of MET (150 mg/kg) for 8 weeks after T2DM modeling, and dihydromyricetin (DHM) group with daily intragastric administration of DHM (250 mg/kg) for 8 weeks after modeling. The levels of fasting blood glucose, low density lipoprotein (LDL-C), triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL-C) and glycosylated hemoglobin (HbA1c) of the rats were measured, and plasma levels of insulin and high mobility group protein-1 (HMGB1) were detected with ELISA. The cardiac function of the rats was assessed using color echocardiography, ECG was measured using a biological signal acquisition system, and myocardial pathology was observed with HE staining. The protein expressions of HMGB1, nuclear factor-κB (NF-κB) p65 and phospho-NF-κB p65 (p-NF-κB p65) in the myocardial tissue were detected using Western blotting.@*RESULTS@#Compared with the control group, the rats in T2DM group showed significant anomalies in cardiac function after modeling with significantly increased plasma HMGB1 level and expressions of HMGB1, NF-κB p65 and p-NF-κB p65 proteins in the myocardial tissue (P < 0.05 or 0.01). Treatment with DHM significantly improved the indexes of cardiac function of the diabetic rats (P < 0.05 or 0.01), decreased plasma HMGB1 level and down-regulated the protein expressions of HMGB1 and p-NF-κB p65 in the myocardial tissue (P < 0.05 or 0.01).@*CONCLUSION@#DHM treatment can improve cardiac function in diabetic rats possibly by down-regulation of HMGB1 and phospho-NF-κB p65 expressions in the myocardium.

Molecular polymorphism Analysis on CD36 Deficiency among Platelet Blood Donors in Shenzhen / 中国实验血液学杂志

OBJECTIVE@#To analyze the molecular polymorphisms of CD36 among 58 blood donors with CD36 deficiency and compare with CD36 positive controls.@*METHODS@#A total of 58 donors with CD36 deficiency during a screening conducted in the laboratory from September 2019 to December 2020 were enrolled as the test group, including 39 males and 19 females, while 120 platelet donors with CD36 positive were randomly selected as the controls, including 76 males and 44 females. All of the subjects were Han nationality. The PCR-SBT method was used to detect coding region of CD36 gene, and molecular mutations were compared with those CD36 positive controls.@*RESULTS@#Among the 58 donors with CD36 deficiency, mutations appears in 32 individuals. The detection rate for type I was 71.43% (5/7), and type II was 51.92% (27/52), while among the 120 controls, mutations appears in 12 donors (10%). In the CD36 antigen-deficient donors, 16 variations were found, in which 329-330 del AC with the highest frequency accounted for 20.69%, followed by 1228-1239 del ATTGTGCCTATT(15.52%) and 1156 C>T(10.34%). Two variations, 198-205 del GATCTTTG and 220 C>T, led to premature termination of translation; four mutations, 329-330 del AC, 560 ins T, 1011-1049 39bp dupl and 1343-1344 ins TCTT, caused translation frame shift; 1228-1239 del ATTGTGCCTATT led to deletion of four amino acids (Ile-Val-Pro-Ile) at sites 410-413 of the peptide chain. The 1140 T>A and 1275 G>A were synonymous mutations, and the other 7 mutations resulted in the substitution of single nucleotide. The platelet expression in the donors of CD36 positive with 329-330 del AC or 1228-1239 del ATTGTGCCTATT mutation (heterozygote) was lower than those CD36 positive individuals without mutations (homozygote).@*CONCLUSION@#Multiple gene mutations in the CD36 coding region may cause CD36 deficiency, and the heterozygous individuals with mutations may lead to CD36 antigen reduction or deletion. Mutation is not detected in 44.83% of CD36 deficient individuals, there may be some other reasons for the CD36 antigen deficiency.

Solutions to cross-matching incompatibility in clinical: retrospective study of 1 770 cases in Shenzhen / 中国输血杂志

【Objective】 To analyze the blood samples sent by hospitals in Shenzhen to solve ABO cross-match incompatibility during 2011 to 2020, so as to find corresponding solutions to improve the efficacy of blood transfusion. 【Methods】 The clinical data of 1 770 cases of cross-match incompatibility in our laboratory from January 2011 to December 2020 were collected and reviewed. The causes of cross-match incompatibility were analyzed, the types of unexpected antibodies were determined. The overall incidence of antibodies was evaluated by statistical method of classified variables. The safety of blood transfusion was safeguarded by ABO homotype plus cross-matching compatibility. 【Results】 1) The 1 770 samples, presenting cross-matching incompatibility, involved 956 patients. The average number of cross-matching per patient from 2011 to 2015 was 1.32(307/232), which increased from 1.27(103/81) in 2016 to 2.23(286/128) in 2018, and remained stable in 2019 and 2020. 2) Among 956 patients, auto-and/or allo-antibody in plasma were yielded in 90.38%(864/956), including auto-antibody plus alloantibody in 42.26%(404/956), solo auto-antibody in 20.71%(198/956) and solo allo-antibody in 27.41%(262/956). Up to 20 kinds of specific allo-antibodies were detected, belonging to 8 blood groups. Among them, 70.82%(551/778) were Rh blood group, such as anti-E(37.15%)>anti-c(20.95%)>anti-C(5.27%)=anti-e(5.27%)>anti-D(2.19%), followed by MNS [11.40%(112/778)], Kidd [5.66%(44/778)], Leiws [3.21%(25/778)], Duffy [1.80%(14/778)], Diego [1.03%(8/778)], P1 [0.39%(3/778)] and H [0.26%(2/778)]. 3) 86%(37/43) of multiple transfusion recipients, aged below 20 years old, were thalassemia, and 1-4 kinds of allo- and/or auto-antibody were yielded. 【Conclusion】 The cross-matching incompatibility were mainly caused by allo- and/or auto-antibodies, which may be induced by blood transfusion, pregnancy or autoimmune diseases such as autoimmune hemolytic anemia.Those suspicious blood samples in clinical should be sent to blood group reference laboratory for further determination, in order to ensure the safety and efficacy of blood transfusion.

Investigation of Volume Load in Outpatients with Hypertension and Analysis of Influencing Factors / 中山大学学报(医学科学版)

@#【Objective】To assess the difference in volume load between different blood pressure control levels in outpatients with hypertension. To investigate the effect of clinical factors on volume load in outpatients with hypertension. To analyze the clinical baseline characteristics and use of antihypertensive drugs in outpatients with hypertension. 【Methods】 A total of 514 outpatients with hypertension were included from July to November 2017. Clinical indicators including gender，age，height，weight，years of hypertension，blood pressure levels，treatment options for hypertension and comorbidities of these patients were recorded. The body volume load was evaluated by detecting the ratio of extracellular fluid（ECW）to total body water（TBW）using a Bioimpedance Analyzer. Whole body ECW/TBW≥0.39 was defined as high volume load. The effects of clinical factors on the volume load of hypertensive patients and whether there was a difference in volume load between different antihypertensive therapy were analyzed. 【Results】The blood pressure compliance rate of outpatients with hypertension was 15.37% ，which was still very low. Male patients had lower blood pressure compliance rate than female patients. Blood pressure was more difficult to control in patients with older age ，higher body mass index （BMI），higher waist-to-hip ratio（WHR）or longer duration of hypertension. The higher the blood pressure grading，the higher the proportion of combination medication. Diuretics were still the most widely used antihypertensive drugs. Age ，gender and different hypertension grades were the main factors affecting the volume load of hypertensive patients. Volume load was higher in female，older or higher systolic blood pressure（SBP）patients. Among them，age was the most important factor affecting the volume load of hypertensive patients.【Conclusion】The blood pressure compliance rate of outpatients with hypertension was still low. Effectively reducing the volume load was one of the important means to control blood pressure，which was more important in female，older or higher SBP patients.

Preliminary investigation of Blomia tropicalis breeding status in ground dust rice in Haikou City / 中国血吸虫病防治杂志

Objective To investigate the Blomia tropicalis breeding status in the ground dust rice collected in Haikou City. Methods Totally 17 samples of ground dust rice were collected from 10 sampling sites in Haikou City. Then 10 g of dust rice was taken from each individual sample for isolation of the mites that were made of slide specimen, and the mites were identified and classified under a microscope. Results B. tropicalis mites were found in all the 17 samples with the detection rate of 100%. A total of 1 176 heads of B. tropicalis were isolated, with an average breeding density of 6.91 heads/g. Conclusion B. tropicalis breeding status is severe in Haikou City. The effective measures should be taken to prevent and control the harm associated with B. tropicalis contamination.

Correlation between LDL-C/HDL-C Ratio and the Severity of Coronary Artery Lesion in Patients with Coronary Heart Disease / 中山大学学报(医学科学版)

[Objective]To explore the correlations between different indices of lipoprotein cholesterol and apolipopro-tein of coronary heart disease(CHD)patients,especially that between low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)and that between apolipoprotein A1(apoA1)and apolipoprotein B100 (apoB100),as well as the correlations between these indices,indices ratios and the severity of coronary artery lesion.[Methods]301 coronary heart disease patients hospitalized to accept percutaneous coronary intervention(PCI)in the Third Affiliated Hospital of Sun Yat-sen University during 2013-2014 were recruited in the study. Fasting serum lipid indices including triglycerides(TG),total cholesterol(TC),LDL-C,HDL-C,apoA1 and apoB100 were examined before surgery.Gensini score was calculated to evaluate the severity of coronary artery lesion. 153 patients whose Gensini score was less than 50 were assigned to Group A,while 148 patients with Gensini score greater than or equal 50 were distributed to Group B.[Results]Positive correlations were found between LDL-C and HDL-C(r=0.161,P=0.005), apoA1 and apoB100(r=0.358,P<0.001),apoB100 and LDL-C(r=0.487,P<0.001),apoA1 and LDL-C(r=0.178, P=0.002)by linear correlation analysis. No significant correlation was found between apoB100 and HDL-C. None of LDL-C,HDL-C,TC was correlated with Gensini score. However,LDL-C/HDL-C ratio was positively correlated with Gensini score(r=0.148,P=0.01). The results showed no significant correlations between apoB100,apoB100/apo A1 ratio and Gensini score but negative correlation between apoA1 and Gensini score(r=-0.129,P=0.025). The positive correlation between HDL-C/LDL-C ratio and Gensini score was still valid after multi-factors adjustment (β=5.071,P=0.018).[Conclusion]Of patients with coronary heart disease,there exist some correlations between LDL-C and HDL-C,apoA1 and apoB100,while the correlation between LDL-C and HDL-C is relatively weak.The LDL-C/HDL-C ratio,weakly positively correlated with the severity of coronary artery lesion,is a risk factor of coronary artery lesion,while the level of apoA1,negatively correlated with the severity of coronary artery lesion,could play a protective role.

Effects of Upper Limb Robot-assisted Therapy on Motor Function and Activities of Daily Living in Patients with Convalescent Stroke / 中国康复理论与实践

Objective To explore the effects of upper limb robot-assisted therapy on motor function and activities of daily living in con-valescent stroke patients. Methods From June to September, 2016, 12 chronic stroke patients at their first-ever stroke were enrolled and ran-domized into experimental group (n=6) and control group (n=6). Both groups received routine rehabilitation. Additional robot-assisted thera-py was provided to the experimental group, and additional repetitive movement training was provided to the control group, 20 minutes a day, five days a week for four weeks. Fugl-Meyer Assessment-Upper Extremities (FMA-UE), modified Ashworth Scale (MAS) and Func-tional Independent Measure (FIM) were used to assess the motor function of the upper limbs and hands, the muscular tension of shoulder and elbow, and activities of daily living (ADL) before and after treatment. Results After treatment, the scores of FMA-UE and FIM were bet-ter in both groups (Z>2.032, P0.05), however, the scores were a little bit higher in the experimental group than in the control group. After treatment, for the experimental group, the MAS scores of shoulder abduction/adduction and elbow flexion/extension improved (Z>2.121, P2.000, P0.05). There was no significant difference in the MAS scores of shoulder abduction/adduction and elbow flexion/extension between two group (Z0.05). The moving trail recorded by the computer, gradually became a regular pattern from the mass, saying the motor control ability became better. Conclusion Upper limb robot-assisted therapy can promote the recovery of the motor function of upper limbs and ADL in convalescent stroke patient, similar to the repetitive movement training.

Development of HPLC-MS/MS method for determination of linezolid in human plasma / 国际药学研究杂志

Objective To determine the concentration of linezolid in human plasma by high performance liquid chromatagra?phy-tandem mass spectrometry(HPLC-MS/MS)so as to provide evidence for the rational administration in clinic. Methods The plas?ma was extracted by acetonitrile with telmisartan as internal standard(IS). The mobile phase was 1 mmol/L ammonium acetate with 0.1%formic acid-acetonitrile with gradient elution. Quantitative analysis of[M+H]+ions was m/z 338.1→m/z 296.3(linezolid)and m/z 515.3→m/z 276.3(telmisartan,the IS). Results Excellent linear relationship was obtained from 0.23 to 46μg/ml with correlation co?efficient value of 0.9993. The lowest limit of quantitation was 46 ng/ml,the extraction recovery was from 87%to 109%. The intra-and inter- day precisions(relative standard deviations)ranged from 2.5% to 8.6%. Conclusion The method is specific,sensitive and simple and can be used to determine the concentration of linezolid in clinic.

Bacterial Succession on Rat Carcasses and Applications for PMI Estimation / 法医学杂志

UNLABELLED@#Abstract:@*OBJECTIVE@#To investigate the bacterial succession on rat carcasses and to evaluate the use of bacterial succession for postmortem interval (PMI) estimation.@*METHODS@#Adult female SD rat remains were placed in carton boxes. The bacterial colonization of circumocular skin, mouth and vagina was collected to be identified using culture-dependent biochemical methods. The changes in community composition were regularly documented.@*RESULTS@#The bacterial succession in three habitats showed that Staphylococcus and Neisseria were predominated in early PMI, especially Staphylococcus aureus and Neisseria lactamica in 6 hours after death. Lactobacillus casei developed on the 3-4 days regularly, and kept stable at a certain level in late PMI.@*CONCLUSION@#The involvement of normal and putrefactive bacteria in three body habitats of rat remains can be used for PMI estimation.

Tuberculous abdominal aortic aneurysm with alimentary tract hemorrhage: a case report with medico-legal implications / 法医学杂志

An autopsy case of sudden death induced by alimentary tract hemorrhage was presented, which was caused by the unexpected rupture of clinically unrecognized tuberculous abdominal aortic aneurysm (TAAA). The initial diagnosis was made of the syndrome of coronary heart disease and hypertensive disease. The detailed autopsy showed that the alimentary tract hemorrhage was caused by a sudden rupture of the mass after posture changing was ascertained as the cause of death. The diagnosis of TAAA was determined by the autopsy findings. Analysis for the medical dispute of TAAA was described, and the difficulty of the diagnosis and medico-legal implications were also discussed.

TGF-β1-induced synthesis of collagen fibers in skeletal muscle-derived stem cells / 华中科技大学学报（医学）（英德文版）

The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells (MDSCs). Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups: group A (control group), group B (treated with TGF-β1, 10 ng/mL), group C (treated with TGF-β1 and anti-connective tissue growth factor (CTGF), both in 10 ng/mL), and group D (treated with anti-CTGF, 10 ng/mL). The expression of CTGF, collagen type-I (COL-I) and collagen type-III (COL-III) in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL-I and COL-III was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL-I and COL-III mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-II induced by TGF-β1 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL-I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL-I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.

TGF-β1-induced synthesis of collagen fibers in skeletal muscle-derived stem cells / 华中科技大学学报（医学）（英德文版）

The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells (MDSCs). Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups: group A (control group), group B (treated with TGF-β1, 10 ng/mL), group C (treated with TGF-β1 and anti-connective tissue growth factor (CTGF), both in 10 ng/mL), and group D (treated with anti-CTGF, 10 ng/mL). The expression of CTGF, collagen type-I (COL-I) and collagen type-III (COL-III) in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL-I and COL-III was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL-I and COL-III mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-II induced by TGF-β1 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL-I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL-I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.

Reflection of estimating postmortem interval in forensic entomology and the Daubert standard / 法医学杂志

Estimating postmortem interval (PMI) is always the emphasis and difficulty in forensic practice. Forensic entomology plays a significant indispensable role. Recently, the theories and technologies of forensic entomology are increasingly rich. But many problems remain in the research and practice. With proposing the Daubert standard, the reliability and accuracy of estimation PMI by forensic entomology need more demands. This review summarizes the application of the Daubert standard in several aspects of ecology, quantitative genetics, population genetics, molecular biology, and microbiology in the practice of forensic entomology. It builds a bridge for basic research and forensic practice to provide higher accuracy for estimating postmortem interval by forensic entomology.

Complications in children after ventriculo-peritoneal shunt / 中华神经医学杂志

Objective To explore the treatments of complications in children after ventriculo-peritoneal shunt (VPS) and its complication appearance reasons.Methods The clinical data of 22 patients occurred complications after VPS,admitted to and performed VPS in our hospital from March 2005 to March 2011,were reviewed retrospectively.The reasons for the complications and treatment results were summarized.Results Obstruction of proximate tube appeared in 6 cases,obstruction of distal end in 4 cases,postoperative infection in 6 cases,subdural hydroma in 4 cases,and silt ventricle syndrome in 2 cases.The short-term response was 955％ after proper treatment.None died for related complications and one child with postoperative infection gave up treatment.Conclusion High risk of complications is noted in children performed VPS; active management based on the reasons of complications can acquire satisfactory curative effects.

The effects of atorvastatin on C-reactive protein induced Toll-like receptor 4 expression on CD14+ monocyte / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of atorvastatin on C-reactive protein (CRP) induced Toll-Like receptor 4 (TLR4)expression on CD14+ monocyte, and the production of proinflammatory cytokines tumor necrosis factor alpha (TNFalpha), interleukin-6 (IL-6), matrix metalloproteinases-9 (MMP-9), and to study the anti-inflammatory mechanisms of statins.</p><p><b>METHODS</b>The monocytes were isolated from blood of healthy volunteers by the Ficoll density gradient and stimulated by CRP with different doses (5, 25, 50, 100 microg/ml) and different exposure time (6, 12, 24, 48 h). Cells were also incubated with atorvastatin of different doses (1.0, 2.5, 5.0, 7.5, 10.0 micromol/L) in the presence of CRP 50 microg/ml. The protein expression of TLR4 was measured by flow cytometry, mRNA expression of TLR4 and of myeloid differentiation protein (MD2)was detected by quantitative PCR. TNFalpha, IL-6, MMP-9 concentrations in supernatants of cultured medium were measured by ELISA.</p><p><b>RESULTS</b>(1) Compared with the un-stimulated control group, enhanced TLR4 protein expression was already detected at a concentration of 5 microg/ml of CRP and increased in a dose-dependent manner (32.22 +/- 2.80)%, (49.94 +/- 5.58)%, (74.82 +/- 3.24)% and (90.82 +/-2.88)% at 5, 25, 50 and 100 microg/ml CRP. (2) TLR4 protein expression on 50 microg/ml CRP stimulated cells also increased in a time-dependent manner (29.80 +/- 2.70)%, (47.44 +/- 4.41)%, (81.71 +/- 2.92)% and (50.57 +/- 3.34)% after 6 h, 12 h, 24 h, 48 h. (3) When monocytes were incubated with CRP 50 microg/ml and atorvastatin (1.0, 2.5, 5.0, 7.5, 10.0 micromol/L), protein expression [(68.17 +/- 1.71)%, (52.43 +/- 1.38)%, (27.72 +/- 4.55)%, (17.46 +/- 3.20)%, (9.99 +/- 2.81)%] and mRNA expression (82.72%, 67.34%, 48.16%, 30.88%, 13.85%) of TLR4 as well as mRNA expression of MD2 (81.78%, 71.04%, 47.85%, 27.06%, 18.30%) were reduced in a dose-dependent manner. (4) Level of TNFalpha, IL-6 and MMP-9 in supernatants was significantly reduced by atorvastatin (2.5 micromol/L) compared with control group (P < 0.01). When monocyte incubated with CRP 50 microg/ml and atorvastatin 10.0 micromol/L, the level of TNFalpha, IL-6, MMP-9 decreased to (25.8 +/- 2.5) microg/ml, (128.2 +/- 14.7) pg/ml, (65.2 +/- 12.3) ng/ml, respectively.</p><p><b>CONCLUSION</b>CRP increased the protein expression of TLR4 on CD14+ monocyte in a dose-dependent and time-dependent manner. Atorvastatin can inhibit the signal transduction of TLR4 and reduce proinflammatory cytokines release induced by CRP on CD14 monocyte, and this might be one of the anti-inflammatory mechanisms of atorvastatin.</p>

Expression levels of RhoA in bronchoalveolar lavage fluid of asthma patients allergic to dust mites / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of RhoA in the bronchoalveolar lavage fluid (BLF) from asthma patients allergic to dust mites and study its clinical significance.</p><p><b>METHODS</b>BLF was collected from 47 mite-allergic asthma patients, 15 asthma patients non-allergic to dust mites and 7 healthy volunteers to detect RhoA expression by immunohistochemistry.</p><p><b>RESULTS</b>The total positivity rates of RhoA expression were 66.7% in mite-allergic asthma patients, 70.2% in asthma patients non-allergic to dust mites and 0 in healthy volunteers, showing significant difference between the 3 groups (P<0.01). No statistical difference was observed in RhoA expression levels between male and female asthma patients allergic to dust mites (P>0.05). The total positivity rate was 83.3% in asthma children allergic to dust mites, higher than that in other patients with different ages, but their RhoA expression levels showed no significant differences (P>0.05). The total positive rates were 33.3%, 57.9%, 88.2% and 100.0% in patients with clinical grades I, II, III and IV, showing significant differences (P<0.05).</p><p><b>CONCLUSIONS</b>The positivity rate of RhoA in the BLF is higher in asthma patients allergic to dust mites than in normal healthy subjects, increases with the clinical grade.</p>