BACKGROUND: The relationship between cellular morphosis and function is undetachable, but there is little investigation about ultrastructure of human umbilical cord mesenchymal stem cells (hUCMSCs).OBJECTIVE: To study the relationship between the functions of hUCMSCs and their ultrastructure obtained by atomic force microscope (AFM). METHODS: hUCMSCs were isolated, cultured, expanded after enzyme digestion. P3 cells were observed under AFM. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified using Oil red O staining and alkaline phosphatase staining. RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for hematopoietic marker CD34. Cells in G_0/G_1 phase accounted for 80%. Proliferation index was 19.9%. Following adipogenic induction, alkaline phosphatase staining demonstrated brown cytoplasm in cube and polygonal cells. AFM showed hUCMSCs were spindle shape, obvious cytoskeletal filament that connected into nets, which fit for the strong capacities for proliferation, migration and differentiation.

Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.

Objective To explore an effective treatment for hypertensive intracerebral hemorrhage.Methods By the method of random and control,patients with hypertensive intracerebral hemorrhage were randomly divided into two groups:the treatment group(32 patients)was treated with integrated traditional Chinese and western medicine therapy,including super-early operation,conventional western medicine and Chuanxiongqin injection treatment.The control group(40 patients)was treated with operation and conventional western medicine treatment.The effect was evaluated on 28 th day after treatment.Results The effect of the treatment group was superior to that of the control group(χ~2=4.15,3.26,P＜0.05).The treatment group had lower mortality rate(χ~2=8.04,P＜0.05)and lower morbidity of complications(pulmonary infection:6/16 cases,χ~2=11.37,P=0.01;upper-congestive hemorrhage:8/16 cases,χ~2=4.10,P=0.04)statistical data indicated that there was significant difference between treatment group and control group.Conclusion Super-early operation with traditional Chinese medicine in treatment of hypertensive intracerebral henorrhage has a better effect than the treatment without traditional Chinese medicine.

Objective To explore the values of diffusion-weighted imaging(DWI)and apparent diffusion coefficient(ADC)measurements in various lesions of multiple sclerosis(MS).Methods Sixty patients with clinically diagnosed remitting-relapsing MS(RRMS)were included to undergo conventional brain MRI and DWI scans.the lesions were included when the diameter was more than 5 mm.mean ADC values were measured for various lesions of MS.The statistical analyses were performed to determine the differences of mean ADC values among various lesions of MS.and to compare the correlation between ADC values of lesions and Expanded Disability Status Scale(EDSS)scores.Results (1)The ADC value of hypointense lesions was significantly higher than that of isointense lesions(F=55.90,P<0.05),the ADC values were(127.5 ±9.3)×10-5mm2/s and(95.7 ±6.3)×10-5mm/s respectively.The nodular enhancing lesions had a significantly lower ADC value than the ring-enhancing lesions(F=64.18,P<0.01).the ADC values were(114.7 ±12.3)×10-5mm2/s and(140.7 ±11.0)×10-5mm2/s respectively.The ADC value of confluent lesions was substantially higher that of discrete lesions(t=9.04,P<0.01).the ADC values were(141.4±6.5)×10-5mm2/s and(105.4±13.9)×10-5mm2/s respectively.(2)No correlation between ADC of lesions and EDSS scores was found(r=0.35,P>0.05).Conclusion DWI and quantitative ADC are useful to explain the pathological changes in different lesions and to monitor the disease duration of MS.

Objective To investigate the brain magnetic resonance imaging(MRI)findings in 33 patients with neuromyelitis optica(NMO).Methods Patients who fulfilled the latest diagnostic criteria of NMO and whose brain MRI did not satisfied with diagnostic criteria of multiple sclerosis(MS)were enrolled.All the patients underwent brain MRI and spinal cord scannings and subsequent images analysis.Results Thirty-three patients with NMO were included to study.Five out of 33(15.2%)patients did not have brain parenchymal abnormalities,28 out of 33 patients(84.8%)were detected to have brain abnormal findings.Brain parenchymal lesions were well-defined in 22 patients(66.7%),no non-specific or atypical brain parenchymal lesions were found in the supratentorium or infratentorium in the other 6 cases(18.2%).However,brain MRI disclosed macroscopic,symmetrical diffuse FLAIR and T2-visible hyperintensity in deep white matter.Fifteen cases had more than one lesion(≥2 lesions),and the other 7 cases had single lesion.Supratentorial lesions were mostly punctate or small dots in nonspecific hyperintensity in juxtacortical,subcortical and deep white matter regions,a few were atypical patches.In the infratentorium,brainstem was an easily involved region(14/33,42.4%),especially in medulla(7/33,21.2%).Conclusions Brain MRI abnormalities are common in Chinese NMO,and brain lesions do not exclude the diagnosis of NMO.The observations of brain lesions are helpful to improve and revise diagnostic criteria of NMO.

OBJECTIVETo study responses of physiological ecology and quality evaluation of Rehmannia glutinosa in continuous cropping.

METHODThe potted plant R. glutinosa which consists of first cropping, 1 year continuous cropping and 2 year continuous cropping were used as experimental materials. The photosynthetic activity, descending axis vitality, the protective enzymes system and MDA content were measured, the quality was evaluated by FTIR and HPLC.

RESULTContinuous cropping reduced the content of chlorophyll in the non-first cropping R. glutinos, the photosynthetic activity and descending axis vitality were weakened. Because of the increase of the free radical in the R. glutinos due to the continuous cropping, the activity of protective enzymes including POD, SOD and CAT were enhanced and MDA content were increased, more importantly the medical potency declined . And along with the increasing years of the continuous cropping, this effect becomes even stronger.

CONCLUSIONContinuous cropping affects the descending axis ability of absorbing water and nutrition and photosynthesis are inhibited R. glutinosa, at the same time, it also causes the disorders of antioxidation systems in R. glutinos, resulting in continuous cropping obstacle and decline of the medicinal materials quality.

Antioxidants ; metabolism ; Ecology ; Photosynthesis ; physiology ; Rehmannia ; enzymology ; growth & development ; metabolism ; physiology ; Superoxide Dismutase ; metabolism

OBJECTIVETo explore effects on allelopathic autotoxicity of Rehmannia glutinosa of different abatement measures.

METHODThe experiments for abating allelopathic effects of R. glutinosa were designed with the reducing rate as a indication, three measures of the treating soil with lime nitrogen, and water washing were employed, R. glutinosa- Achyranthes bidentata rotation system and using Achyranthes bidentata as green manure, and seedling transplant were also studied for the abatement effect.

RESULTEffect of seedling transplant was the best, reducing rate of transplant and bare root transplant on the 20th day after emergence was 76.80%, 71.70%, respectively. The reducing rate of the treating soil with microorganism reached 54.25%. The effects of water washing and R. glutinosa-A. bidentata rotation system and using A. bidentata as green manure were not satisfied. Especially only using A. bidentata as green manure without rotation system worsened the allelopathic effects of R. glutinosa.

CONCLUSIONAll measures can abate allelopathic effects of R. glutinosa to some extent except only using A. bidentata as green manure.

Crops, Agricultural ; Rehmannia ; growth & development ; toxicity ; Seedlings ; physiology ; Soil Microbiology

Effects of the leaf-clipped treatment on growth and physiological properties of Rehmannia glutinosa were studied. Result showed that with the increase of the cutting leaves degree, growth rates of shoots and roots were decreased, sugar contents declined, chlorophyll contents decreased and root activities also inhibited. Compared with the normal plant (CK), the root inhibit rates in T1, T2, T3 were 17.53%, 33.41%, 59.47%, respectively. Physiological indexes including chlorophyll contents, root activities and sugar contents also were impacted by the leaf-clipped treatment. The results indicate that to balance source-sink relationship is a very essential method for improving production of R. glutinosa.
Agriculture ; Carbohydrate Metabolism ; Carbohydrates ; analysis ; Chlorophyll ; analysis ; metabolism ; Plant Leaves ; chemistry ; growth & development ; physiology ; Rehmannia ; chemistry ; growth & development ; physiology

This experiment was designed to investigate the feasibility of transplanatation of using porous PHB as cell carrier for the transplanatation of adrenocortical cells. Adrenocortical cells from rat adrenal gland were separated and cultured in vitro. The effect of PHB on the proliferation and secretory function of adrenocortical cells were evaluated by MTT and RIA methods. Then adrenocortical cells were seeded into porous PHB. After the cells were cultured in vitro for about seven days, they were implanted into the rats having undergone bilateral adrenalectomy. The changes of blood corticosterone and aldosterone and the local histological changes in these rats were observed. Adrenocortical cells were able to grow and survive on PHB. No effect on the proliferation and secretory function of adrenocortical cells were observed. Most bilateral adrenalectomized rats bearing the transplanted adrenocortical cells within PHB (study group) survived longer than did the adrenalectomized rats in control group. The blood corticosterone level and aldosterone level of study group were higher than those of control group. It was found that PHB has no effects on the survival, proliferation and secretory function of adrenocortical cells. Adrenocortical cells within PHB can survive a period of time and can secrete corticosterone and aldosterone which can meet the needs of the adrenalectomized rats. PHB can degrade slowly in vivo. It is feasible to perform transplantation of adrenocortical cells using porous PHB as cell carrier.
Adrenal Cortex ; cytology ; Aldosterone ; blood ; urine ; Animals ; Biodegradation, Environmental ; Cell Proliferation ; Cell Transplantation ; Cells, Cultured ; Coculture Techniques ; Corticosterone ; blood ; urine ; Hydroxybutyrates ; chemistry ; pharmacology ; Male ; Polyesters ; chemistry ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo explore the molecular mechanism of continuous monoculture problem by constructing the cDNA libraries of continuous monoculture Rehmannia glutinosa.

METHODTo use the suppression subtractive hybridization (SSH) technique to construct the forward and reverse subtractive cDNA libraries of continuous monoculture R. glutinosa to adopt blue-white colony screening and PCR to detect the positive clones which would be sequenced and analyzed by bioinformatics.

RESULTThe subtracted cDNA libraries of continuous monoculture R. glutinosa. were successfully constructed, and the result showed that the forward and reverse subtracted libraries obtained 300 positive clones, respectively. The forward and reverse libraries got different ESTs, and produced 232 (forward library) and 214 (reverse library) unique ESTs by sequencing. Based on homology search of BLASTX and BLASTN in NCBI, 200 and 195 of unique ESTs were homologous to known genes in the forward and reverse libraries, respectively. Categories of orthologous group (COG) showed that the forward and reverse libraries got 60 and 61 ESTs with the corresponding gene annotation, involving 21 metabolic pathways.

CONCLUSIONThe information of differential expression genes in continuous monoculture R. glutinosa, and their functional annotation of differentially expressed genes indicate that continuous monoculture has a profound effect on expression of the genes in R. glutinosa. Furthermore, the research analyzed several key genes in response to replant problem, which provided a foundation for revealing the molecular mechanism of continuous monoculture R. glutinosa.

Expressed Sequence Tags ; Gene Expression Profiling ; Gene Expression Regulation, Plant ; Gene Library ; Polymerase Chain Reaction ; Rehmannia ; genetics ; metabolism ; Sequence Analysis, DNA