OBJECTIVETo explore the effect of Bushen Huoxue Compound (BHC) on lactate dehydrogenase (LDH) leakage, expressions of vascular endothelial growth factor (VEGF) and VEGF mRNA in retinal Muller cells under high glucose condition or advanced glycosylation end products (AGEs) condition by using serum pharmacological method.

METHODSThe retinal Müller cells of 5-7 days post-natal Sprague Dawley (SD) rats were cultured with modified enzyme-digestion method. Purified retinal Muller cells were cultured in normal conditions, high glucose condition (50 mmol/L) or AGEs (50 mg/L and 100 mg/L) conditions, and BHC-containing serum was added to culture medium. The LDH leakage and VEGF expressions were measured by enzyme-linked immunosorbent assay (ELISA). In addition, the relative expression of VEGF mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the normal control group, expressions of VEGF and VEGF mRNA were significantly increased in the high glucose group, the low dose AGEs group and the high dose AGEs group (all P < 0.01). The LDH leakage was obviously increased in the high dose AGEs group, when compared with the normal control group and the high glucose group (P < 0.01). The LDH leakage, expressions of VEGF and VEGF mRNA were obviously decreased by BHC-containing serum both in high glucose and AGEs conditions (P < 0.05, P < 0.01). BHC-containing serum had no significant effect on the LDH leakage and expressions of VEGF and VEGF mRNA in normal conditions (P > 0.05).

CONCLUSIONSAGEs intervention could obviously lower the stability of Müller cell membrane. Up-regulated expressions of VEGF and VEGF mRNA in cultured Müller cells could be induced by AGEs or high glucose. BHC-containing serum could stabilize the stability of Müller cell membrane, inhibit the transcription of VEGF mRNA and decrease the protein expression of VEGF, which might be one of important mechanisms for preventing and treating diabetic retinopathy.

Animals ; Cells, Cultured ; Diabetic Retinopathy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Ependymoglial Cells ; Glucose ; L-Lactate Dehydrogenase ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A

Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of human spermatogenetic mechanism.

Objective:To explore the effect of the microenvironment induced by damaged mouse hepatic cells on the conversion of human umbilical cord blood-derived cells into hepatocyte-like cells. Methods: A hepatic injury-like microenvironment was mimicked using carbon tetrachloride damaged mouse hepatic cells, where mononuclear cells (MNC) from human umbilical cord blood were cultured in a compartment separated by trans-well membrane. Histochemical staining, reversed transcription-polymerase chain reaction (RT-PCR) and gene sequencing were performed for the information on the conversion of human umbilical cord blood MNC. Results: A number of PAS positive stained cells in MNC co-cultured with damaged mouse hepatic cells were observed after 72 h. Cells expressing mature hepatocyte markers, human albumin (hALB) and human GATA-4 (hGATA-4) mRNA were detected by RT-PCR, which was further confirmed with sequencing. Relevant control groups, MNC co-cultured with normal mouse hepatic cells and MNC cultured alone remained negative. Conclusion: The culture system using damaged mouse hepatic cells as stimulator could be a potential in vitro system for the conversion of human umbilical cord blood-derived cells into hepatocyte-like cells.

OBJECTIVETo observe the effect of Pinggan Qianyang Recipe (PQR) on inhibiting angiotensin II (Ang II) induced proliferation and migration of vascular smooth muscle cells (VSMCs) and changes of DNA methylation.

METHODSVSMCs were cultured using tissue explant method, and PQR containing serum was prepared. Primarily cultured VSMCs were divided into four groups, the normal group, the model group, the folate group (folic acid intervention) , and the PQR group. The proliferation and migration of VSMCs was duplicated by Ang II. After 24-h Ang II induced culture, 40 microg/mL folic acid was added to the folate group for 48 h, while 5% PQR containing serum was added to the PQR group for 48 h. The cell growth curve of VSMCs was drawn by using Cell Counting Kit (CCK-8). The proliferative activity of VSMC was determined by MTT assay. The migration of VSMCs was measured by Millicell chamber. The general level of cytosine methylation in cell nucleus was detected via 5-mC antibodies immunofluorescence, and mRNA expression levels of DNA methyltransferase 1 (DNMT1) were measured by Real-time q-polymerase chain reaction (q-PCR).

RESULTSVSMCs were promoted by Ang II at 10(-6) mol/L for 24 h. Compared with the normal group, the proliferative activity and migration quantity of VSMCs obviously increased, and DNA methylation level obviously decreased (P < 0.05, P < 0.01). Compared with the model group, the cell growth, proliferative activity and migration quantity of VSMCs obviously decreased and the general DNA methylation level increased in the folate group and the PQR group (P < 0.05, P < 0.01). Compared with the normal group, the mRNA expression of DNMT1 decreased in the model group (P < 0.01). Compared with the model group, mRNA expression of DNMT1 in Ang II induced VSMCs was obviously enhanced in the folate group and the PQR group (P < 0.01).

CONCLUSIONSPQR could inhibit Ang II induced proliferation and migration of VSMCs, and cause high genomic DNA methylation level. Changes of DNA methylation might be associated with DNMT1 expression.

Angiotensin II ; pharmacology ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects

BACKGROUND: Increasing studies have shown that cell-sheet engineering is a more promising method for the treatment of myocardial infarction. OBJECTIVE: To construct sandwich-like myocardial cell sheets with electrospun chitosan-collagen membrane as support membrane and with adipose-derived stem cells as seed cells. METHODS: In this study, we prepared electrospun chitosan-collagen membranes at different mass ratio (7:3, 5:5) as a support membrane. Adipose-derived stem cells at passage 4 that were cultured in a thermo-sensitive petri dish for 3 days were seeded onto the two kinds of electrospun membranes at 20 ℃ for 15 minutes until the cell sheets rolled up. Then, the electrospun membranes and cell sheets with the cell layers facing up were placed together onto another thermo-sensitive petri dish in which adipose-derived stem cells were confluent completely. After 30 minutes of culture, the cell sheets were cultured in myocardial cell medium. Two weeks later, cell morphology was observed using multi-photon microscopy. Immunocytochemistry, western blot and flow cytometry were used to detect the expression of TnI and Cx43 in the myocardium. Real-time quantitative PCR was used to detect cardiomyocyte-specific gene expression. RESULTS AND CONCLUSION: From the results of scanning electronic microscopy (SEM) and propidium iodide (PI) staining, we could found that ADSCs grew easily on the chitosan-collagen (5:5) membrane, and there were more dead cells on the chitosan-collagen (7:3) membrane than on the chitosan-collagen (5:5) membrane. After 2 weeks of differentiation with the myocardial cell medium, higher troponin I and Cx43 protein expressions were observed on the chitosan-collagen (5:5) membrane (P＜0.05). Moreover, the mRNA levels of α-skA, β-MHC, TnI, Cx43, ANP, GATA-4 and Nkx2.5 were higher in the chitosan-collagen (5:5) membrane group than the chitosan-collagen (7:3) membrane group. All these results indicate that the chitosan-collagen (5:5) membrane is better for the growth and differentiation of adipose-derived stem cells, as well as for sandwich-like cell-sheet construction.

OBJECTIVETo study the clinicopathologic features of different variants of primary focal segmental glomerulosclerosis (FSGS).

METHODSOne hundred and two cases of FSGS were retrieved from the archival files of Peking University First Hospital during the past 6-year period. The pathologic findings were reviewed and the degrees of active and chronic changes were assessed by morphometric analysis. The histopathologic patterns were then correlated with clinical manifestations.

RESULTSAmongst the 102 cases of primary FSGS studied, 55.9% belonged to the NOS (not other specified) variant, while the perihilar, cellular, tip and collapsing variants accounted for 6.9%, 25.5%, 4.8% and 6.9% respectively. The level of proteinuria in the cellular and tip variants were much higher than that in the NOS variant; and the incidence of nephrotic syndrome in the tip and collapsing variants was higher than that in the other three variants (chi(2) = 12.23, P < 0.05). The activity score of the cellular and collapsing variants was also higher than that of the other three variants (P < 0.05). The interval between disease onset and renal biopsy diagnosis in the perihilar variant was longer than that in the other variants. The chronicity score of this variant was higher than that of the tip and NOS variants (P < 0.05). On the other hand, the total scores of active and chronic changes of the tip variant was lower than that of the cellular and collapsing variants (P < 0.05); and its chronic score was lower than that of the NOS and perihilar variants (P < 0.05).

CONCLUSIONSThe NOS variant is the commonest morphologic pattern seen in primary FSGS. The cellular and collapsing variants are the patterns associated with active lesions, while perihilar variant is the pattern associated with chronic lesions. The tip variant shows mild pathological changes compared with the other patterns.

Adolescent ; Adult ; Creatinine ; blood ; Female ; Glomerulosclerosis, Focal Segmental ; blood ; classification ; pathology ; Humans ; Kidney Glomerulus ; pathology ; Male ; Serum Albumin ; metabolism ; Young Adult

The aim of this study was to investigate the expression of macrophage migration inhibitory factor (MMIF),hypoxia-inducible factor-1 α (HIF-1 αt) and vascular endothelial growth factor (VEGF) in the serum and endometrial tissues of patients with endometriosis (EM) and the clinical significance.Eighty EM patients [American Reproductive Association stage I (n=20),stage Ⅱ (n=22),stage Ⅲ (n=21) and stage Ⅳ (n=17)] were enrolled and divided into mild (10-14 points,n=28),moderate (16-24 points,n=27) and severe (26-30 points,n=25) dysmenorrhea groups.The control group included 40 healthy women of childbearing age who underwent routine healthcare examinations in the enrolment period.The expression of MMIF,HIF-1α and VEGF in the serum and endometrial tissues was measured by enzyme-linked immunosorbent assay and Western blotting,respectively.Meanwhile,the sensitivity and specificity of serum MMIF,HIF-1α,and VEGF when separately used as single indexes or jointly used as one index were examined as well.The results showed that serum concentrations of MMIF,HIF-1α,and VEGF were significantly higher in EM patients than in controls (P＜0.05).The expression of all three proteins in both serum and endometrial tissues increased significantly with the R-AFS stage (P＜0.05) and with dysmenorrheal severity (P＜0.05).The sensitivity and specificity of the combined detection of serum MMIF,HIF-1α,and VEGF levels were significantly higher than those of single index detection (P＜0.05).In conclusion,the expression of MMIF,HIF-1α,and VEGF in the serum and endometrial tissues may be used to assess the stage of EM and the severity of dysmenorrhea.Combined evaluation of MMIF,HIF-1α,and VEGF significantly improves the diagnostic sensitivity and specificity.

Either cetuximab or bevacizumab can improve the survival of patients with metastastic colorectal cancer (mCRC) if administered combided with cytotoxic agents. However, the effect of two or more target agents in combination is uncertain in these patients. Here, we reported a patient with mCRC successfully treated by a combination of target agents after the failure of chemotherapy. The patient received palliative resection of primary tumor followed by 9 cycles of postoperative XELOX regimen, cytokine-induced killer cell (CIK)-based biotherapy, traditional Chinese medicine, particle implantation in the lung metastatic lesions. The tumor progressed 20 months after the standard treatments. Then, the regimen cetuximab, bevacizumab and cefitinib was applied. During the treatment with targeted agents, grade IV acne-like rash and relatively severe parionychia of the toes occurred. Both of them recovered smoothly. The PET-CT reexamination at 40 days after the target treatment showed that the metabolism of mediastinal lymph nodes basically recovered to a normal level. The combination of multiple targeted agents obtained a progression-free survival(PFS) of 11 months and the patient with a good quality of life during this period.
Adenocarcinoma ; diagnostic imaging ; drug therapy ; pathology ; secondary ; Angiogenesis Inhibitors ; therapeutic use ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bevacizumab ; Catheter Ablation ; Cetuximab ; Cytokine-Induced Killer Cells ; immunology ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Drug Delivery Systems ; Fluorouracil ; analogs & derivatives ; therapeutic use ; Humans ; Immunotherapy, Adoptive ; Liver Neoplasms ; secondary ; surgery ; Lung Neoplasms ; secondary ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Multimodal Imaging ; Neoplasm Staging ; Positron-Emission Tomography ; Quality of Life ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Sigmoid Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; Tomography, X-Ray Computed

OBJECTIVETo investigate the analgesic and sedative effects of inhaling a mixture of nitrous oxide and oxygen on burn patient during and after dressing change.

METHODSA total of 240 burn patients hospitalized in the Institute of Burn Research of Changhai Hospital Affiliated to the Second Military Medical University, Department of Burns of the First People's Hospital in Zhengzhou, and Department of Burns and Plastic Surgery of General Hospital of Ningxia Medical University from October 2011 to September 2012 were enrolled in our study, and they were all in accordance with the inclusion criteria. The 240 patients were divided into control group (n = 60, treated with inhalation of oxygen during dressing change) and treatment group (n = 180, treated with inhalation of a mixture of 65% nitrous oxide and oxygen during dressing change) according to the computer-generated list of random number. The other treatments in control group and treatment group were the same. Before, during, and after dressing change, heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), oxygen saturation (SO2), and adverse effects were observed. The degree of pain and anxiety felt by the patients were respectively evaluated with the visual analogue scale (VAS) and Chinese version of the burn specific pain anxiety scale (C-BSPAS) at the same time points as above. Data were processed with analysis of covariance, chi-square test, analysis of variance, and rank sum test.

RESULTSThere were no significant differences between control group and treatment group in the levels of HR, SBP, DBP, and SO2 before dressing change (with F values respectively 0.76, 0.06, 1.11, 0.70, P values all above 0.05). Compared with those of control group, the levels of HR, SBP, DBP, and SO2 in treatment group were significantly ameliorated during dressing change (with F values respectively 81.78, 146.36, 226.44, 205.62, P values all below 0.01). After dressing change, the levels of DBP in the two groups were close (F = 0.31, P > 0.05), but the levels of HR, SBP, and SO2 showed statistical differences (with F values respectively 7.02, 8.69, 12.23, P < 0.05 or P < 0.01). Before dressing change, the VAS scores were approximate between control group and treatment group (Z = 0.21, P > 0.05). Compared with those in control group (9.4 ± 0.7, 1.7 ± 2.5), the VAS scores were significantly lowered in treatment group during and after dressing change (1.6 ± 1.3, 0.7 ± 1.1, with Z values respectively 11.84, 3.35, P values all below 0.01). There was no significant difference in C-BSPAS score between control group and treatment group before dressing change (Z = 0.62, P > 0.05). Compared with those in control group (75 ± 13, 73 ± 12), the C-BSPAS scores in treatment group were decreased during and after dressing change (9 ± 15, 9 ± 14, with Z values respectively 11.91, 12.28, P values all below 0.01). There were no obvious adverse effects in two groups before, during, and after dressing change.

CONCLUSIONSA mixture of nitrous oxide and oxygen seems to have obvious analgesic and sedative effects on burn patients during dressing change, and it can be widely used.

Administration, Inhalation ; Adolescent ; Adult ; Aged ; Analgesia ; methods ; Bandages ; Burns ; surgery ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Nitrous Oxide ; administration & dosage ; therapeutic use ; Oxygen ; administration & dosage ; therapeutic use ; Young Adult

Liver cancer is a common malignant tumor in China and a major health concern. We aimed to estimate the liver cancer incidence and mortality in China in 2010 using liver cancer data from some Chinese cancer registries and provide reference for liver cancer prevention and treatment. We collected and evaluated the incidence and mortality data of liver cancer in 2010 from 145 cancer registries, which were included in the 2013 Chinese Cancer Registry Annual Report, calculated crude, standardized, and truncated incidences and mortalities, and estimated new liver cancer cases and deaths from liver cancer throughout China and in different regions in 2010 from Chinese practical population. The estimates of new liver cancer cases and deaths were 358,840 and 312,432, respectively, in China in 2010. The crude incidence, age-standardized rate by Chinese standard population (ASR China), and age-standardized rate by world standard population (ASR world) were 27.29/100,000, 21.35/100,000, and 20.87/100,000, respectively; the crude, ASR China, and ASR world mortalities were 23.76/100,000, 18.43/100,000, and 18.04/100,000, respectively. The incidence and mortality were the highest in western regions, higher in rural areas than in urban areas, and higher in males than in females. The age-specific incidence and mortality of liver cancer showed a rapid increase from age 30 and peaked at age 80-84 or 85+. Our results indicated that the 2010 incidence and mortality of liver cancer in China, especially in undeveloped rural areas and western regions, were among high levels worldwide. The strategy for liver cancer prevention and treatment should be strengthened.
China ; epidemiology ; Female ; Humans ; Incidence ; Liver Neoplasms ; epidemiology ; mortality ; Male ; Registries ; Rural Population ; Sex Distribution ; Urban Population