Influence of enteral feeding initiation time on intestinal flora and metabolites in very low birth weight infants: a prospective study / 中国当代儿科杂志

OBJECTIVES@#To study the influence of enteral feeding initiation time on intestinal flora and metabolites in very low birth weight (VLBW) infants.@*METHODS@#A total of 29 VLBW infants who were admitted to the Department of Neonatology, Children's Hospital of Chongqing Medical University, from June to December, 2020, were enrolled as subjects. According to the enteral feeding initiation time after birth, the infants were divided into two groups: <24 hours (n=15) and 24-72 hours (n=14). Fecal samples were collected at weeks 2 and 4 of hospitalization, and 16S rDNA high-throughput sequencing and gas chromatography-mass spectrometry were used to analyze the microflora and short-chain fatty acids (SCFAs) respectively in fecal samples.@*RESULTS@#The analysis of microflora showed that there was no significant difference between the two groups in Chao index (reflecting the abundance of microflora) and Shannon index (reflecting the diversity of microflora) at weeks 2 and 4 after birth (P>0.05). The analysis of flora composition showed that there was no significant difference in the main microflora at the phylum and genus levels between the two groups at weeks 2 and 4 after birth (P>0.05). The comparison of SCFAs between the two groups showed that the <24 hours group had a significantly higher level of propionic acid than the 24-72 hours group at week 4 (P<0.05), while there was no significant difference in the total amount of SCFAs and the content of the other SCFAs between the two groups (P>0.05).@*CONCLUSIONS@#Early enteral feeding has no influence on the diversity and abundance of intestinal flora in VLBW infants, but enteral feeding within 24 hours can increase the level of propionic acid, a metabolite of intestinal flora.

Paeoniae Rubra Radix decreases the hepatotoxicity of Psoraleae Fructus in an immunologically stressed rat model: a metabolic network analysis / 药学学报

An immunologically stressed rat model was used in a metabolomics study on the ability of Paeoniae Rubra Radix to reduce the liver toxicity of Psoraleae Fructus. Different groups of rats were given the extracts of Psoraleae Fructus and Psoraleae Fructus together with Paeoniae Rubra Radix or combined with a non-toxic dose of lipopolysaccharide (LPS). The biochemical indices of liver function and pathological changes in liver tissue were used to evaluate histopathological changes. UHPLC-QTOF/MS was used to analyze the metabolic profile of serum samples, combined with principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) methods. The HMDB database and Metabo Analyst online tool were used for biomarker identification and metabolic pathway-enrichment analysis. The results show that the co-treatment Psoraleae Fructus and LPS resulted in significant liver injury, indicated by the elevation of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities, as well as obvious pathological changes. Liver injury was significantly decreased by treatment with Paeoniae Rubra Radix. Metabolomic analysis showed that the addition of Paeoniae Rubra Radix ameliorated the abnormal serum metabolism in rats mainly through regulation of arachidonic acid metabolism and glycerophospholipid metabolism pathways.

Effect of Dihydrotestosterone on CostimulatoryMolecules in a Mouse Model of Graves' Disease / 中国医学科学杂志(英文版)

Objective Graves' disease is the most common autoimmune thyroid disease and its prevalence and clinical manifestations are disparate between females and males. Costimulatory molecules play an essential role in regulating autoimmune responses. The objective of this study was to determine if expression of inhibitory molecules was correlated with treatment by dihydrotestosterone (DHT) in an BALB/c mouse model of experimental autoimmune Graves' disease.Methods Female BALB/c mice were immunized three times with thyroid stimulating hormone receptor A-subunit encoded by adenovirus to establish a Graves' disease model. Three different doses of DHT or a matching placebo were administered by implantation of slow-release pellets a week before the first immunization. Four weeks after the third immunization, the mice were euthanatized, and then the spleen and thymus were removed. Total thyroxine and free thyroxine levels in serum of mice were detected using a radioimmunoassay kit. Real-time polymerase chain reaction was performed to estimate the expression of costimulatory molecules in lymphocytes from the spleen and thymus. Flow cytometry was used to analyze the percentage of CD4 T cells in splenic lymphocytes. Quantitative data were compared with unpaired -tests. Correlation between two variables was analyzed using Analysis of Variance.Results Treatment with DHT can dramatically reduce total thyroxine and free thyroxine levels. Higher expression of programmed death-1 was found in the spleen of Graves' disease mice receiving 5 mg of DHT treatment (0.635±0.296 . 0.327±0.212; =2.714, =0.014), similarly, T-cell immunoglobulin domain and mucin domain 3 (TIM-3) in both the spleen (1.004±0.338 . 0.646±0.314; =2.205, =0.022) and the thymus (0.263±0.127 . 0.120±0.076; =3.221, =0.004) also increased after 5 mg of DHT treatment compared with the parallel placebo model mice. Moreover, the percentage of CD4 T cells declined in the splenic lymphocytes of Graves' disease mice treated with 5 mg of DHT (19.90%±3.985% . 24.05%±2.587%; =2.804, =0.012). A significant negative association was observed between expression of TIM-3 in the spleen and serum levels of total thyroxine (=-0.7106, =0.014) as well as free thyroxine (=-0.6542, =0.029).Conclusion This study demonstrates that DHT can ameliorate experimental autoimmune Graves' disease, which may occur by up-regulating expression of programmed death-1 and TIM-3 and inhibiting development of CD4 T cells.

A new phenylpropanoid glycoside from leaves of Platycladus orientalis / 中草药

Objective: To study the chemical constituents from the leaves of Platycladus orientalis, as well as their antioxidant and α-glucosidase inhibitory activities. Methods: The compounds were isolated and purified by silica gel, MCI, polyamide, and prep-HPLC chromatography, their structures were elucidated by spectral analysis. DPPH and ABTS methods were used to study the antioxidant activity, and pNPG method was used to study the α-glucosidase inhibitory activity. Results: Eleven compounds (1-11) were isolated from the 80% ethanol extract of the leaves of P. orientalis, and identified as 4-O-(1’,3’-dihydroxypropan-2’-yl)- dihydroconiferyl alcohol 9-O-β-D-glucopyranoside (1), myricetrin (2), 5,8,3’,4’-tetrahydroxy-flavone-7-O-β-D-xylopyranoside (3), isomassonianoside B (4), (-)-isopramine 9’-O-β-D-glucopyranoside (5), (7R,8S,7’S,8’R)-4,9,4’,7’-tetrahydroxy-3,3’-dimethoxy- 7,9’-epoxylignan 4-O-β-D-glucopyranoside (6), sugiol (7), totarol (8), 5,6-dehydrosugiol methyl ether (9), isopimara-8,15-dien-7-one (10) and ethanol α-L-rhamnopyranoside (11). Conclusion: Compound 1 is a new compound, named as platycloside A; In the known compounds, seven compounds (4-7, 9-11) are isolated from this plant for the first time, six compounds (4-6, 9-11) are isolated from the family Thujoideae for the first time, and four compounds (5, 6, 10, 11) are isolated from the family Cupresaceae for the first time. The compounds 2-6 showed a degree of antioxidant activities. The compounds 2 and 3 showed the α-glucosidase inhibitory activities.

Research Progress on Effect of Traditional Chinese Medicine on Blood-brain Barrier Permeability / 中国实验方剂学杂志

The blood-brain barrier is located between blood vessels and brain parenchyma and is a composite tissue composed of brain capillary endothelial cell,astrocyte end foot,pericyte,basement membrane and their tight connections.The blood-brain barrier acts as an important barrier between the blood-brain. It strictly restricts the exchange of blood and brain tissue. On the one hand,it allows the nutrients required by the brain tissue to enter the barrier. On the other hand,the substance that damages the brain tissue is restricted from entering the barrier to maintain the stability of the neuron microenvironment. The barrier function of the blood-brain barrier also limits the concentration of drugs that treat certain diseases into the brain or reduces the concentration of drugs into the brain,affecting the treatment of certain diseases.Studies have found that Chinese medicine has the effect of regulating the permeability of the blood-brain barrier. For example, Chinese herbal medicines such as Moschus,borneol,Styorax,and Benzoinum can increase the permeability of the blood-brain barrier,and can help its medicine into the brain. Its role is mainly related to the reduction of the blood-brain barrier tight junction,the inhibition of the blood-brain barrier transporter,and the inhibition of the active transport of ion channels.Orifice-opening medicinal such as borneol and Moschus can reduce the permeability of the blood-brain barrier.Certain tonifying and replenishing medicinal such as Ginseng Radix et Rhizoma,Astragali Radix,and Paeoniae Radix Alba can also reduce the permeability of the blood-brain barrier to protect the blood-brain barrier and brain tissue.Its role is mainly related to the regulation of inflammatory reactions,increased expression of tight junction-associated proteins,and promotion of vascular endothelial proliferation.The two-way regulation of blood-brain barrier permeability by traditional Chinese medicine(TCM) may be the basis for the prevention and treatment of brain diseases by TCM.In this paper,we systematically sort out the relevant literatures on the study of the permeability of blood-brain barrier and its mechanism of action in Chinese medicine in recent years,and carry out in-depth combing and summary,in order to provide a scientific basis for the clinical application of TCM to prevent and treat brain diseases.

A new sesquiterpene glycoside from Osmanthus fragrans var. aurantiacus / 药学学报

The chemical constituents were isolated and purified by column chromatography and semi-preparative reversed-phase high performance liquid chromatography with silica gel, MCI and polyamide in order to study the chemical constituents of dried flowers of Osmanthus fragrans var. aurantiacus. Their structures were identified by the physical and chemical properties and one-dimensional nuclear magnetic resonance (1H-, 13C-NMR, DEPT), two-dimensional nuclear magnetic resonance (1H-1H COSY, non-decoupled HSQC, HSQC, HMBC), UV, IR and high resolution mass spectrometry data. One new compound (1) and five known compounds (2-6) were isolated from 95% ethanol extract of dried broccoli. They were identified as (9S)-9-hydroxymengastigm-5-en-4-one-9-O-primeveroside (1), oleanolic acid (2), forsythiaside (3), 2-(4-hydroxyphenethyl)-ethanol-(6-acetyl)-β-D-glucopyranoside (4), salidroside (5), and acteoside (6). Compounds (2-6) were isolated from this plant for the first time.

Development of Predictive Quantitative Retention-Activity Relationship Models of Quinolones by Mixed Micellar Liquid Chromatography / 中国药学杂志

OBJECTIVE: To obtain the adequate QRAR models of the half-life (t1/2), clearance(CL), volume of distribution (Vd) and area under concentration-time curve (AUC) of quinolones and elucidate the advantages and limitations of using mixed micellar liquid chromatography for describing and estimating the biological parameters. METHODS: The BMCBrij35/SDSQRAR models using mixed micellar system of Brij35/SDS (85:15 ) as a mobile phase under adequate experimental conditions were developed for the biological parameter estimation of quinolones. The correlation between retention factors and biological activities was investigated using second order polynomial models. The predictive and interpretative ability of the chromatographic models was evaluated in terms of cross-validated data (RMSEC, RMSECV and RMSECVi). RESULTS: The BMCBrij35/SDSQRAR models of t1/2, CL, Vd and AUC were statistically significant and both interpolation and extrapolation of parameters were reasonably adequate. CONCLUSION: The mixed micellar liquid chromatography can simulate the resting membrane potential and the conformation of the long hydrophilic polyoxyethylene chains, which may become a simple, economic, and highly reproducible option for establishing QRAR model.

Effect of microecological therapy on rotavirus diarrhea and immunologic function in infants / 基础医学与临床

Objective To investigate the effect of microecological therapy on rotavirus ( RV)-induced diarrhea and infants’ immunologic function in infants. Methods 150 infants with RV diarrhea were randomly divided into the ex-perimental group and the control group. The control group was given the treatment of montmorillonite powder. The experimental group was given live combined bifidobacterium and lactobacillus tablets in addition to montmorillonite powder. The clinical efficacy and the improvement of clinical symptoms were compared between the two groups. RV antigen and immunologic function were detected with colloidal gold method. Immune function was detected by flow cytometry and ELISA. Results The total effective rate of the experimental group was 90.7%,and it was significant-ly higher than that of the control group which was 74.7%(P＜0.05). Compared with the control group, the antidi-arrheal time, fever clearance time and disappearance time of vomiting symptoms in the experimental group were shorter ( P＜0.05) . The negative conversion rate of RV virus antigen in the experimental group after 1 and 2 weeks treatment was significantly higher than that in the control group ( P＜0.05) . After 3 weeks treatment, RV virus antigen in the experimental group all turned negative, but the control group still had a positive rate of 12.0% ( P＜0.05) . The peripheral blood CD3+, CD4+and the ratio of CD4+/CD8+in the experimental group after treatment were significantly higher than those before treatment and those in the control group after treatment ( P＜0.05) , the same phenomenon was found in serum immunoglobulin A ( IgA) and IgM ( P＜0.05) . No adverse events occurred during the treatment period in the two groups. Conclusions Microecological therapy has an exact curative effect and high safety in the treatment of infantile RV diarrhea, it could effectively improve the immunologic function of the infants.

Clinical effect of OPD-Scan Ⅲ marker methods in Toric intraocular lens implantation / 眼科新进展

Objectivc To compare the accuracy and practicability of OPD-Scan Ⅲ marker methods and slit light narrow band marker methods in phacoemulsification and Toric IOL implantation.Methods A prospective randomized controlled study was conducted in 70 eyes of 70 patients with regular astigmatism (1.00-3.00 D).All patients underwent phacoemulsification and Toric IOL implantation and were randomly divided into two groups according to preoperative marker methods:observant group,in which OPD-Scan Ⅲ marker methods were used in 35 eyes of 35 patients,and control group,with application of slit light narrow band marker methods in 35 eyes of 35 patients.The postoperative visual acuity,residual astigmatism and deviation of lens axis (LAD) were observed and compared between the two groups.Results The uncorrected visual acuity (UCVA) between preoperative data and 3 months after surgery was statistically significant [(0.11 ± 0.09) vs.(0.59 ± 0.25)] (P =0.000).The difference in the postoperative 1 day and 3 months LAD was no statistically significant (P ＞ 0.05).The astigmatism between preoperation and 3 months after treatment was significant [(1.88 ± 0.49) D vs.(0.54 ± 0.30) D] (P =0.000).There was no significant difference in LAD between the two groups neither the first day nor 3 months after operation (both P =0.621).Conclusion OPD-Scan Ⅲ marker methods and slit light narrow band marker methods are both highly accurate and practical,but the former requires less for operator and is more easily accepted by patients.

Acute massive gastrointestinal bleeding in the elderly

An 82-year patient presented with nausea, coffee-ground emesis, melena and hematochezia on July 10, 2011. The patient received blood transfusion on July 11, and continued to bleed from July 12 to 17. The patient underwent upper gastrointestinal endoscopy on July 16. There were no abnormalities in the esophagus, stomach and duodenum. Then the patient presented with shortness of breath, extreme fear, fatigue, hypotension, sweating, and cold limbs. Dopamine, as well as pressurised infusion of packed red blood cells and fresh frozen plasma were given to the patient to maintain blood pressure. CT angiography [CTA] revealed no aortic fistula, and enteroscopy revealed active bleeding in the vicinity of the ligament of Treitz. The retrograde exploration of gastroscopy revealed a 5×4 cm diverticulum on the posterior wall of the duodenum under the ligament of Treitz. Active bleeding of the small artery in the diverticulum was observed via incision of the duodenal wall, and the diverticulum was isolated. Hemostasis was achieved after ligation of blood vessels, and diverticulectomy was performed. Enteroscopy is important for the diagnosis of duodenal and upper small intestinal diseases. Repeated endoscopic exploration of multiple sites in the small intestine revealed the cause of the bleeding. The multidisciplinary team finally found the cause of the bleeding and its proper management

The Lung Function Impairment in Non-Atopic Patients With Chronic Rhinosinusitis and Its Correlation Analysis

OBJECTIVES: Chronic rhinosinusitis (CRS) is common disease in otorhinolaryngology and will lead to lower airway abnormality. However, the only lung function in CRS patients and associated factors have not been much studied. METHODS: One hundred patients with CRS with nasal polyps (CRSwNP group), 40 patients with CRS without nasal polyps (CRSsNP group), and 100 patients without CRS were enrolled. The difference in lung function was compared. Meanwhile, CRSwNP and CRSsNP group were required to undergo a bronchial provocation or dilation test. Additionally, subjective and objective outcomes were measured by the visual analogue scale (VAS), 20-item Sino-Nasal Outcome Test (SNOT-20), Lund-Mackay score, Lund-Kennedy endoscopic score. The correlation and regression methods were used to analyze the relationship between their lung function and the above parameters. RESULTS: The forced expiratory volume in 1 second (FEV1) and forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75) of CRSwNP group were significantly lower than other groups (P<0.05). On peak expiratory flow, there was no difference between three groups. In CRSwNP group, FEV1 was negatively correlated with peripheral blood eosinophil count (PBEC) and duration of disease (r=–0.348, P=0.013 and r=–0.344, P=0.014, respectively), FEF25-75 negatively with VAS, SNOT-20 (r=–0.490, P=0.028 and r=–0.478, P=0.033, respectively) in CRSsNP group. The incidence of positive bronchial provocation and dilation test was lower in CRSwNP group (10% and 0%, respectively), with both 0% in CRSsNP group. The multiple linear regression analysis indicated that change ratio of FEV1 before and after bronchial provocation or dilation test were correlated with PBEC in CRSwNP group (β=0.403, P=0.006). CONCLUSION: CRS leading to impaired maximum ventilation and small airway is associated with the existence of nasal polyp. Lung function impairments can be reflected by PBEC, duration, VAS, and SNOT-20. In CRSwNP patients, PBEC is independent predictor of FEV₁ change ratio.

Expression of CD163 in children with Epstein-Barr virus infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical significance of CD163 in the diagnosis and the evaluation of severity and prognosis of childhood hemophagocytic lymphohistiocytosis (HLH).</p><p><b>METHODS</b>Ninety-four children were classified into Epstein-Barr virus (EBV)-positive (n=55) and EBV-negative groups (n=39; control group). The EBV-positive group was subgrouped into infectious mononucleosis (IM; n=47) and HLH (n=8). Serum levels of soluble CD163 were measured using ELISA. Expression of CD163 on mononuclear cells was detected by flow cytometry.</p><p><b>RESULTS</b>The serum levels of soluble CD163 were>10 000 ng/mL in all eight HLH patients (>30 000 ng/mL in 3 cases). The mean serum levels of soluble CD163 in the HLH group were significantly higher than in the control and IM groups (P<0.05). The serum levels of soluble CD163 in EBV-positive children were positively correlated with EBV-DNA copies and serum levels of ferritin and LDH, but were negatively correlated with white blood cell count, neutrophil count, hemoglobin and platelet count. The follow-up after treatment for three HLH patients showed that serum levels of soluble CD163 were significantly reduced, but the soluble CD163 levels rebounded in one patient who was complicated by fungal pneumonia infection.</p><p><b>CONCLUSIONS</b>The levels of serum soluble CD163 may be related to the severity in children with HLH. The EBV-positive children with soluble CD163 levels >10 000 ng/mL should be considered the possibility of HLH.</p>

Influence of recombinant lentiviral vector encoding miR-15a/16-1 in biological features of human nasopharyngeal carcinoma CNE-2Z cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the influence of recombinant lentiviral vector encoding miR-15a/16-1 on biological features of human nasopharyngeal carcinoma CNE-2Z cells and underlying mechanisms.</p><p><b>METHODS</b>GFP-positive CNE-2Z cells transfected with recombinant lentiviral vector were selected. The experiment was divided into control group, transfected group, radiotherapy group, transfected-radiotherapy group. Cell proliferation was analyzed by MTT. Apoptosis was detected by flow cytometry. Radiotherapy sensitivity of the cells in control group and transfected group was evaluated by colony forming experiment. The expressions of miR-15a, miR-16-1 and bcl-2 mRNA were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The expression of bcl-2 protein was detected by Western blot. The activation of Caspase-2 and Caspase-3 was evaluated by spectrophotometry.</p><p><b>RESULTS</b>Relative expression quantities of miR-15a and miR-16-1 in infected group were 524.80 ± 40.79 (t = 494.611, P = 0.000) and 466.11 ± 40.96 (t = 386.8, P = 0.000), respectively. The proliferation of the cells in transfected-radiotherapy group was the most obvious, followed by the cells in radiotherapy group and transfected group (F = 424.3, P = 0.000). The apoptosis rates of control group, transfected group, radiotherapy group and transfected-radiotherapy group were (2.2 ± 1.4)%, (9.6 ± 0.8)%, (2.9 ± 1.1)%, and (18.6 ± 0.7)% respectively(F = 158.5, P = 0.000). Clonogenic assay showed that the values of SF2, Do (1.473) and Dq (1.581) in transfected group were lower than those in control group. The relative expression levels of bcl-2 mRNA in transfected group, radiotherapy group, and transfected-radiotherapy group had no significant difference (P > 0.05). Decrease in the expression of bcl-2 protein in transfected-radiotherapy group was most significantly, followed by that in transfected group. The percentages of activated Caspase-2 in control group, radiotherapy group, transfected group and transfected -radiotherapy group were 0.12 ± 0.01, 0.24 ± 0.04, 0.35 ± 0.02, and 0.44 ± 0.04, respectively (F = 115.500, P = 0.000). The percentages of activated Caspase-3 in the groups were 0.13 ± 0.01, 0.27 ± 0.01, 0.43 ± 0.02, and 0.83 ± 0.06, respectively (F = 439.921, P = 0.000).</p><p><b>CONCLUSIONS</b>Recombinant lentiviral vector LV-miR15a/16-1 could improve the expression of miR-15a and miR-16-1 in CNE-2Z cells, inhibit the proliferation of CNE-2Z cells, promote apoptosis and enhance the sensitivity of the cells to radiotherapy probably by inhibiting bcl-2 expression, activating Caspase-2 and Caspase-3.</p>

Clinical variables affecting the pregnancy rate of intracervical insemination using cryopreserved donor spermatozoa: a retrospective study in China

The aim of this study was to investigate whether several clinical variables can affect the pregnancy rate of intracervical insemination [ICI] using cryopreserved donor spermatozoa. In this retrospective study, age, years of infertility, cervicitis, urinary luteinizing hormone [LH] surge, insemination number, uterus position, endometrial thickness and morphology, maximal follicle diameter, and the number of dominant follicles on the day of human chorionic gonadotropin [HCG] administration were retrospectively analyzed in 501 women who underwent their first ICI cycle using cryopreserved donor spermatozoa. Increased age, length of infertility [>5 years], retroverted uterine position, and endometrial thickness [<7 mm or >14 mm] were associated with lower rates of pregnancy. In older women with infertile periods longer than five years, especially those with a retroverted uterus, intrauterine insemination [IUI] combined with ovarian stimulation should be recommended. In vitro fertilization with donor spermatozoa [IVFD] should be offered earlier to achieve a much higher success rate.

Detecting phospho-signaling protein of bone marrow leukemia cells by phospho-signaling flow cytometry / 中国实验血液学杂志

The purpose of this study was to establish the phospho-specific flow cytometry (phospho-flow) to detect the phosphorylated signaling proteins of leukemia cells and to evaluate its useful value in leukemia study. The bone marrow of leukemia children was collected, and treated by phospho-flow of extracted mononuclear cells (MNC) and phospho-flow of directly fixed bone marrow (BM) respectively. In phospho-flow of extracted MNC, the MNC extracted from BM were fixed and permeabilized, then were cultured with P-AKT and P-ERK1/2, finally were analyzed by flow cytometry. In phospho-flow of directly fixed BM, the BM was treated with fixation/lysis buffer and 90% methanol, then were incubated with the surface CD antibody, P-AKT and P-ERK1/2, finally the treated BM cells were analyzed by flow cytometry. The results showed that the positive rates of P-AKT and P-ERK1/2 in MNC treated by phospho-flow of extracted MNC of 26 leukemia children were 46.2% and 30.8% respectively, while the positive rates of P-AKT and P-ERK1/2 in BM treated by phospho-flow of directly fixed BM were 50.0% and 38.5% respectively. The comparison of positive rates of P-AKT and P-ERK1/2 between the 2 treatment protocol showed no difference (p > 0.05). It is concluded that the phospho-flow of directly fixed BM established by our laboratory can be used to analyze the signaling proteins of leukemia cells.

Change of CD4(+) CD25(+) regulatory T cells and NK Cells in peripheral blood of children with acute leukemia and its possible significance in tumor immunity / 中国实验血液学杂志

This study was purposed to investigate the changes of CD4(+) CD25(+) regulatory T cells and NK cells in peripheral blood of acute leukemia children at different stages, the function of immune system and the possible roles of the CD4(+) CD25(+) regulatory T cells as well as NK cells in leukemia immunity. The number and proportion of CD4(+) CD25(+) regulatory T cells and NK cells were detected by flow cytometry in the peripheral blood of 53 acute leukemia children, including 25 patients in new diagnosis and 28 patients in continuous complete remission (CCR), and were compared with that of 20 normal children. The results indicated that the mean proportion of CD4(+) CD25(+) CD127(+) in CD4(+) T cells of peripheral blood in newly diagnosed patients, patients with CCR and normal children were (9.55 +/- 2.41)%, (8.54 +/- 2.51)% and (6.25 +/- 0.85)% respectively, the mean proportions of CD4(+)CD25(+)CD127(+) in newly diagnosed patients and patients with CCR were higher than that in normal children, the mean proportion of CD4(+)CD25(+)CD127(+) in newly diagnosed patients were higher than that in patients with CCR (p < 0.05). At the same time, the NK cell count in patients with acute leukaemia decreased as compared with normal control, while after achieving CCR, the NK cell count in patients were also less than that in normal control (4.11 +/- 3.87% and 10.41 +/- 7.20% vs 14.06 +/- 5.95%, p < 0.05). It is concluded that the application of CD4(+), CD25(+) and CD127(+) to detect regulatory T cells is a simple, reproductive and accurate method, and the CD4(+) CD25(+) CD127(+) T cells can better reflect the proportion of CD4(+)CD25(+) regulatory T cells. The increase of regulatory T cells and decrease of NK cells in pediatric patients with acute leukemia indicate that the function of NK cells may be depressed. Treg T cells play a role in occurrence and development of leukemia, and are involved in down-regulating NK cell function.

Test of transient expression of inducible nitric oxide synthase gene transferred to rabbit periodontal ligament cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This article tests a recombinant plasmid pEGFP-iNOS encoding human inducible nitric oxide synthase (iNOS) transient expression in rabbit periodontal ligament cells, and this may contribute to transfer iNOS gene to animal periodontal tissue in vivo.</p><p><b>METHODS</b>Rabbit periodontal ligament cells were transfected with pEGFP-iNOS by means of lipofectamine media methods. Transient transfection were evaluated by fluorescent microscope. After transferring 12 h and 48 h, the transcription of iNOS gene was tested by quantitative real-time PCR and the expression product of pEGFP-iNOS was identified by Western blot. Blank plasmid-transfected group and non-transfected group were used as control.</p><p><b>RESULTS</b>The expression of green fluorescence protein was detected in 12 h after transferring. The transcription of iNOS mRNA and expression of iNOS protein in cells transfected with pEGFP-iNOS were detected by real-time PCR and Western blot.</p><p><b>CONCLUSION</b>The iNOS gene was transfected successfully, the exogenetic iNOS mRNA can be correctly transcribed and expressed in rabbit periodontal ligament cells. In vivo gene transfer of iNOS to rabbit periodontal tissue is feasible.</p>

A proteomic analysis of human dental pulp cells undergoing odontoblast differentiation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To analyze the difference in protein profiles between human dental pulp cells (DPC) and odontogenic differentiated DPC by using proteomic approach.</p><p><b>METHODS</b>Human DPC were induced to odontoblast differentiation and total proteins in the cell lysates before and after induction were prepared. Proteins spots were isolated by two-dimensional gel electrophoresis. DeCyder V6.0 software was applied to gel image analysis. Differential protein spots were identified by peptide mass fingerprinting technique.</p><p><b>RESULTS</b>Forty-six protein spots were determined to be differentially expressed with twenty identified protein spots. Expression changes of the identified proteins revealed the involvement of various regulation mechanisms in odontoblast differentiation, such as cell cycle, cellular energy regulation and signal transduction.</p><p><b>CONCLUSIONS</b>The proteomic approach is a high throughput method to screen the candidate proteins involved in odontoblast differentiation of DPC.</p>

A differential expression proteomic study of human periodontal ligament cell during osteogenic differentiation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To obtain regulating proteins during human periodontal ligament cell (hPDLC) osteogenic differentiation and investigate its underlying molecular mechanism.</p><p><b>METHODS</b>Two-dimension difference in gel electrophoresis (2D-DIGE) coupled with matrix-assisted laser adsorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis was used to identify regulating proteins during hPDLC osteogenic differentiation.</p><p><b>RESULTS</b>A differently expressed protein profile was obtained 7 days after osteogenic induction, and 61 protein spots with significant differences, 29 protein spots were identified by MALDI-TOF-MS, including cytoskeleton, cell membrane-bounding, nuclear regulation, matrix synthesis, and metabolic enzymes and signal transduction and other proteins. A functional class of cytoskeleton proteins showed co-regulation, these proteins were intimately involved in the reorganization of the cytoskeleton, cytokinesis and migration during PDLC differentiation.</p><p><b>CONCLUSIONS</b>A database of differently displayed proteins during hPDLC osteogenic differentiation was established, which may be helpful to understand and further study molecular mechanism of PDLC osteogenic differentiation.</p>

Biological characteristics of bone marrow-derived mesenchymal stem cells in children with acute leukemia / 中国实验血液学杂志

This study was aimed to investigate the conditions of culturing in vitro mesenchymal stem cells (MSCs) derived from bone marrow of children with acute leukemia and the biological characteristics of MSCs from leukemia children. The bone marrow MSCs of acute leukemia children were isolated by density gradient centrifugation combined with adherent segregating method and cultured in DMEM/F12. The morphology of Wright stained MSCs was observed under inverted microscope. Cell surface markers were analyzed with flow cytometry. The growth characteristic features of cultured MSCs was measured with MTT method. Induced adipogenic and osteogenic differentiation of MSCs in appropriate induction media was observed. The results indicated that BM-MSCs of acute leukemia children could be successfully cultured in vitro in appropriate conditions. At 24 hours of culture the MSCs began to adhere to wall, grew in colony and appeared in different shapes. As the culture lasted, the MSCs proliferated continuously and shaped in fusiform. After 2 - 3 weeks of culture, MSCs covered the bottom of culture flask. The analysis of growth feature showed that MSCs were in latency for 3 days, and then entered into growth period. After 8 days of culture the growth of MSCs showed to be in plateau stage. The shape of MSCs in 1st and 2nd generation showed to be heterogeneous but the 3rd generation to be homogeneous with long-fusiform. Cells were arranged in shape of whirlpool or radiation. The surface marker analysis showed that the MSCs were positive for CD105, CD29, CD13, but negative for CD34, CD45, CD14 and HLA-DR. The MSCs from leukemia children could be induced into adipocytes and osteocytes in appropriate conditions. It is concluded that (1) MSCs derived from children with acute leukemia can be successfully cultured and passaged in vitro; (2) MSCs from leukemia children not received chemotherapy are more successfully cultured in vitro than those received chemotherapy; (3) the common biological characteristics of MSCs from children with acute leukemia are same as the MSCs from healthy person.