BACKGROUND: Using bone marrow mesenchymal stem cells (MSCs) infected by an adenoviral vector cen-ying human hepatocyte growth factor (HGF) gene, which was independently developed by our laboratory and has been approved by SFDA in clinical trial to repair bone defects via cell therapy.OBJECTIVE: To investigate the bone restoration effect of mssenchymal stem cell and hepatocyte growth factor in the pathophysiological course of femoral head osteonecresis defects.DESIGN, TIME AND SETTING: An in vivo experiment of ceUular-rnatedal science. The experiment was performed at the Beijing Institute of Radiation Medicine and the 66400 Orthopaedic Hospital of Chinese PLA from September to December 2007. MATERIALS: Eighteen New Zealand rabbits with clean grade, aged 26-28 weeks, were supplied by Beijing Kaiyuan rabbit livestock farm. Bone matrix gelatin was purchased from Shanghai Xiaobo Technological Development Limited Co., Ltd. METHODS: MSCs from New Zealand white rabbit were isolated and culture-expanded by adhesion method and their in vitro ostengenesis and adipogenesis were identified. Eighteen New Zealand white rabbits were created femoral heads defect models and randomly divided into 3 groups, with 6 animals in each group. Animals in each group were treated with scaffold of bone matrix gelatin (27 mm3 per defect) only, scaffolds seeded with MSCs (1×107 per defect) or MSCs infected by an adenoviral vector carrying human hepatocyte growth factor gene (MSC/HGF, 1×107 per defect), respectively. Histological examination was conducted at 3 months post operation. MSCs or MSC/HGF were labeled with carboxyfluorescein diacetate succinmidyl ester dye, treated with cobalt chloride for 72 hours and their proliferative status was evaluated and compared by flow cytomatric techniques. MAIN OUTCOME MEASURES: The differentiation of MSCs, therapeutic effect of MSCs in treating femoral head necrosis, and potent ability of MSCs/HGF against hypoxia.RESULTS: The adherent cells could differentiate into osteoblasts and adipocytes under in vitro inductive condition. Histological examination revealed that the bone defects from both control and MSCs-treated groups were fille:d with fibrous tissue, though blood vessels were evident in MSCs-treated group, whereas new bony tissues were obvious in MSC/HGF group. The result was further confirmed by Lane-Sandhu scaling, which indicated that new bone formation was more evident in MSC/HGF-treated group compared with MSCs-treated or control group (P< 0.01). Flow cytometry analysis showed that the proportion of MSC/HGF-treated group that had experienced cell division was significantly higher than that of MSCs-treated group after cobalt treatment (P < 0.001).CONCLUSION: MSC/HGF exhibit greater osteogeneeis in vivo in this model compared their counterparts, which might be attributed to their resistance to hypoxic injury. The results here suggest that HGF gene modification might be an optional strategy for the application of MSCs in the management of avascular osteonecrosis.

Study the serum level of HGF, Cys C and TGF-beta1 in type 2 diabetic nephropathy (DN), the infect of Pingshen decoction on those index. Selected 69 cases of 2 type DN and randomly divided into therapy group (36 cases) and control group (33 cases). The therapy group were treated with Pingshen decoction 1 dose/d, bid po. The control group were treated with NephritisShu tablet, 6 tablet, tid po. 8 weeks was a course. Before and after treatment, we examine the serum level of HGF, Cys C and TGF-beta1 by ELISA and immunonephelometry, and compare with 30 cases of healthy control group. The study demonstrates that before treatment, the serum level of HGF in both groups were significantly lower than healthy control group (P < 0.01), but Cys C, TGF-beta1 were significantly higher (P < 0.01). After treatment, the serum level of HGF of both groups were increased. The serum level of HGF of therapy group were significantly higher than of control group (P < 0.01), but the serum level of Cys C and TGF-beta1 were significantly lower than control group (P < 0.01). The serum level of HGF was correlated negatively with Cys C,TGF-beta1. In control group, the UAER, urine beta2-MG and quantity of 24-hour urine protein were significantly decreased after treatment (P < 0.01). The index of urine of therapy group were significantly lower than control group (P < 0.01). Results indicate that test of serum level of HGF and Cys C,TGF-beta1 of diabetic nephropathy have important clinical significance. Pingshen decoction can effectively intervene in the serum level of HGF and Cys C, TGF-beta1 and index of urine.
Aged ; Aged, 80 and over ; Case-Control Studies ; Cystatin C ; blood ; Diabetic Nephropathies ; blood ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hepatocyte Growth Factor ; blood ; Humans ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo learn the normal values of exhaled nitric oxide (eNO) in children.

METHODSchool children in Beijing from 11 to 18 years of age were included in the study. All the students were assigned into two groups: normal group and abnormal group (with allergic disease) according to the International Study of Asthma and Allergy in Childhood questionnaires. eNO, peak expiratory flow rate and sensitization were measured.

RESULTTotally 395 students were screened out as normal subject (male: 177, female: 218). The eNO level was not significantly different between genders (P > 0.05), but was associated positively with age in both male and female group (P = 0.008 and P = 0.05 respectively) and associated with height in male students (P = 0.02). The geometric mean value of eNO was 11.22 ppb (parts per billion, ppb = 10(9)) in children aged from 11 to 14 years and 14.13 ppb in children aged from 14 to 18 years, with 95% confidence interval 4.17 - 30.20, 5.50 - 36.31 ppb. The eNO level was significantly increased in children who "ever had asthma or wheezing" (n = 68), and children who "ever had rhinitis" (n = 96) compared with normal subjects (P = 0.001 and P = 0.008). The geometric mean value of eNO was 16.98 ppb in children with positive skin prick test and was significantly increased as compared with children with negative skin prick test with eNO level at 11.75 ppb (P = 0.001).

CONCLUSIONeNO level varied between 10.72 ppb and 13.80 ppb in normal children 11 - 18 years of age, and was positively associated with age and height, but not with gender. eNO level increased significantly in children with wheezing and atopy.

Adolescent ; Asthma ; physiopathology ; Case-Control Studies ; Child ; China ; Exhalation ; physiology ; Female ; Humans ; Male ; Nitric Oxide ; analysis ; physiology

OBJECTIVETo explore the effect of ginkgo leaf extract (GLE) on vascular endothelial function (VEF) in patients with early stage diabetic nephropathy (DN).

METHODSSixty-four patients were randomized equally by a randomzing digital table into two groups, the treated group and the control group. They were all treated for 8 weeks with conventional therapy for diabetes, but GLE tablets were given to the treated group additionally. Changes in VEF were estimated before and after treatment by ultrasonic examination of the brachial artery. In the meantime, changes in plasma levels of the von Willebrand factor (vWF), nitric oxide (NO) and endothelin-1 (ET-1) were observed as well.

RESULTSThe brachial arterial endothelium dependent dilating function in the treated group increased from 4.91+/-2.31% before treatment to 6.78+/-3.89% after treatment (P<0.05), while the level of vWF decreased from 182.05+/-64.13% to 128.56+/-48.98%, and that of NO increased from 50.16+/-24.64 micromol/L to 70.65+/-28.71 micromol/L (P<0.01). However, these indexes were not significantly changed in the control group after treatment (P>0.05).

CONCLUSIONGLE could decrease the plasma level of vWF, raise the plasma NO level and improve the endothelium dependent vascular dilating function in DN patients.

Aged ; Brachial Artery ; drug effects ; pathology ; Diabetic Nephropathies ; blood ; drug therapy ; physiopathology ; Endothelium, Vascular ; drug effects ; Female ; Ginkgo biloba ; chemistry ; Humans ; Male ; Middle Aged ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Plant Leaves ; chemistry

OBJECTIVETo investigate the hematological characteristics of co-inheritance of α-thalassemia (α-thal) and β-thalassemia (β-thal) and to survey the incidence of co-inheritance of α-thal and β-thal in Guangxi.

METHODSDNA samples from 370 primary and middle school students who were β-thal carriers in Guangxi were further processed for the α-goblin gene mutation screening, and were grouped based on the genotype of β- and α-goblin gene. The hematological indexes to the different groups were compared by One-way ANOVA.

RESULTSOf the total 370 β-thal carriers, 79 were found to carry α-thal, which gave a frequency of 21.35% for β-thal carriers and 1.36% for coincidence of these two common disorders in the local population. As expected, the 79 patients presented very variable α-globin alterations in combination with β-globin mutations, showing 31 genotype combined with the coincidence of both Hb disorders. Except the genotypes of 3 β-thal heterozygotes combined with ααα(anti3.7) triplication and 2 β-thal carriers with IVS-II-654(C→T)/N combined-α(3.7)/αα presented the phenotype of thalassemia intermedia, and other 74 carriers with co-inheritance of α-thal and β-thal all presented the phenotype of β-thal trait. There were significant differences between β-thal heterozygotes and the carriers with a co-inheritance of both β+α(0) thal in MCH, MCV and Hb. In addition, there existed significant difference between the carriers with a co-inheritance of both β+α(+) thal and a co-inheritance of both β+α(0) thal in MCV, MCH and Hb.

CONCLUSIONCompared to that of β-thal heterozygotes, the carriers with a co-inheritance of α-thal and β-thal had slighter phenotype with hematological characteristics. It's difficult to distinguish the double heterozygotes with the co-inheritance of α-thal and β-thal from β-thal heterozygotes by hematological indexes, the molecular diagnosis should be performed.

Child ; China ; epidemiology ; Female ; Gene Frequency ; Genotype ; Humans ; Incidence ; Male ; alpha-Thalassemia ; blood ; epidemiology ; genetics ; beta-Thalassemia ; blood ; epidemiology ; genetics

A method of ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field Obitrap high-resolution mass spectrometry(UHPLC-Q-Exactive MS) was established to comprehensively identify the metabolites of carnosic acid in rats. After oral gavage of carnosic acid CMC-Na suspension in rats, urine, plasma and feces samples were collected and pretreated by solid phase extraction(SPE). Acquity UPLC BEH C_(18 )column(2.1 mm×100 mm, 1.7 μm) was used with 0.1% formic acid solution(A)-acetonitrile(B) as the mobile phase for the gradient elution. Biological samples were analyzed by quadrupole/electrostatic field Obitrap high-resolution mass spectrometry in positive and negative ion mode. Based on the accurate molecular mass, fragment ion information, and related literature reports, a total of 28 compounds(including carnosic acid) were finally identified in rat samples. As a result, the main metabolic pathways of carnosic acid in rats are oxidation, hydroxylation, methylation, glucuronide conjugation, sulfate conjugation, S-cysteine conjugation, glutathione conjugation, demethylation, decarbonylation and their composite reactions. The study showed that the metabolism of carnosic acid in rats could be efficiently and comprehensively clarified by using UHPLC-Q-Exactive MS, providing a reference for clarifying the material basis and metabolic mechanism of carnosic acid.
Abietanes ; Animals ; Chromatography, High Pressure Liquid ; Mass Spectrometry ; Rats ; Solid Phase Extraction

Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.

The metabolites of salvianolic acid A and salvianolic acid B in rats were analyzed and compared by ultra-high-perfor-mance liquid chromatography with linear ion trap-orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS). After the rats were administrated by gavage, plasma at different time points and urine within 24 hours were collected to be treated by solid phase extraction(SPE), then they were gradient eluted by Acquity UPLC BEH C_(18) column(2.1 mm×100 mm, 1.7 μm) and 0.1% formic acid solution(A)-acetonitrile(B) mobile phase system, and finally all biological samples of rats were analyzed under negative ion scanning mode. By obtaining the accurate relative molecular mass and multi-level mass spectrometry information of metabolites, combined with the characteristic cleavage law of the reference standard and literature reports, a total of 30 metabolites, including salvianolic acid A and B, were identified. Among them, there were 24 metabolites derived from salvianolic acid A, with the main metabolic pathways including ester bond cleavage, dehydroxylation, decarboxylation, hydrogenation, methylation, hydroxylation, sulfonation, glucuronidation, and their multiple reactions. There were 15 metabolites of salvianolic acid B, and the main biotransformation pathways were five-membered ring cracking, ester bond cleavage, decarboxylation, dehydroxylation, hydrogenation, methylation, sulfonation, glucuronidation, and their compound reactions. In this study, the cross-metabolic profile of salvianolic acid A and B was elucidated completely, which would provide reference for further studies on the basis of pharmacodynamic substances and the exploration of pharmacological mechanism.
Animals ; Benzofurans ; Caffeic Acids ; Chromatography, High Pressure Liquid ; Lactates ; Mass Spectrometry ; Rats ; Technology

OBJECTIVE: To investigate the involvement of endothelial cells (ECs)-derived exosomes in the anti-apoptotic effect of Danhong Injection (DHI) and the mechanism of DHI-induced exosomal protection against postinfarction myocardial apoptosis. METHODS: A mouse permanent myocardial infarction (MI) model was established, followed by a 14-day daily treatment with DHI, DHI plus GW4869 (an exosomal inhibitor), or saline. Phosphate-buffered saline (PBS)-induced ECs-derived exosomes were isolated, analyzed by miRNA microarray and validated by droplet digital polymerase chain reaction (ddPCR). The exosomes induced by DHI (DHI-exo), PBS (PBS-exo), or DHI+GW4869 (GW-exo) were isolated and injected into the peri-infarct zone following MI. The protective effects of DHI and DHI-exo on MI hearts were measured by echocardiography, Masson's trichrome staining, and TUNEL apoptosis assay. The Western blotting and quantitative reverse transcription PCR (qRT-PCR) were used to evaluate the expression levels of miR-125b/p53-mediated pathway components, including miR-125b, p53, Bak, Bax, and caspase-3 activities. RESULTS: DHI significantly improved cardiac function and reduced infarct size in MI mice (P<0.01), which was abolished by the GW4869 intervention. DHI promoted the exosomal secretion in ECs (P<0.01). According to the results of exosomal miRNA microarray assay, 30 differentially expressed miRNAs in the DHI-exo were identified (28 up-regulated miRNAs and 2 down-regulated miRNAs). Among them, DHI significantly elevated miR-125b level in DHI-exo and DHI-treated ECs, a recognized apoptotic inhibitor impeding p53 signaling (P<0.05). Remarkably, treatment with DHI and DHI-exo attenuated apoptosis, elevated miR-125b expression level, inhibited capsase-3 activity, and down-regulated the expression levels of proapoptotic effectors (p53, Bak, and Bax) in post-MI hearts, whereas these effects were blocked by GW4869 (P<0.05 or P<0.01). CONCLUSION DHI and DHI-induced exosomes inhibited apoptosis, promoted the miR-125b expression level, and regulated the p53 apoptotic pathway in post-infarction myocardium.
Mice ; Animals ; Tumor Suppressor Protein p53/metabolism* ; Endothelial Cells/metabolism* ; Exosomes/metabolism* ; bcl-2-Associated X Protein/metabolism* ; Myocardium/metabolism* ; Myocardial Infarction/drug therapy* ; Apoptosis ; MicroRNAs/metabolism*