Residual behaviors and metabolic pathway of ethylparaben in Drosophila melanogaster.

OBJECTIVE: Parabens are commonly used as preservatives in foodstuffs, cosmetics, and pharmaceutical products. The widespread use of parabens has led to their leaking into the environment. Concerns about the safety of parabens have recently increased due to their potential endocrine-disrupting effects as an emerging contaminant. Thus, it is necessary to study the metabolism of parabens in vivo. METHODS: In this study, Drosophila melanogaster in males and females were exposed to ethylparaben (EP) concentration group (300 mg/L, 700 mg/L, and 1000 mg/L), and control group (0 mg/L) by the capillary feeding assay (CAFE). We quantified the activity of the detoxification-related carboxylesterase (CarE). The contents of EP metabolites in D. melanogaster, including p-hydroxybenzoic acid (PHBA), methylparaben (MP), and intact EP were carried out by high-performance liquid chromatography (HPLC). The regression model between EP metabolites (PHBA and MP) and CarE was developed using the Fourier series fitting method. RESULTS: The general level of EP metabolites (PHBA, MP, and intact EP) accumulation was accounted for 5.6-11.5% in D. melanogaster. As EP accumulated, the activity of CarE increased, and the activity of CarE in females was higher than males, which is inconsistent with the result of EP intake dose. Additionally, there were significant differences in the proportion of EP metabolites between female and male flies, and the results of sex comparison were different depending on the EP treated groups and EP metabolites. In general, PHBA of EP hydrolytic product and MP of EP transesterification product in D. melanogaster were 41.4-63.9% and 10.4-24.6%, respectively. In terms of the rest of the EP existed in intact form and ranged from 22.4% to 34.0%. Moreover, the EP metabolites in the conjugated form were higher than those in the free form. The regression model between EP metabolites and CarE was established, showing that the CarE activity can be used to estimate the content of PHBA and MP. CONCLUSION: The result indicates that the EP can accumulate in the body through food. Hydrolysis is the main metabolic pathway of EP in D. melanogaster, and transesterification is another metabolic pathway of EP. Additionally, the EP metabolites in flies mainly exist in conjugated form. Furthermore, the Fourier series fitting method model between EP metabolites and CarE, providing theoretical support to study the dose-effect relationship between metabolites of parabens and CarE. This study not only provides a mathematical basis for the safety evaluation of parabens, but also provides support for the further study of the toxicological effects of parabens.

Hypoxia-inducible factor prolyl hydroxylase inhibitor roxadustat (FG-4592) protects against renal ischemia/reperfusion injury by inhibiting inflammation.

Hypoxia-induced inflammation is the critical pathological feature of acute kidney injury (AKI). Activation of hypoxia-inducible factor (HIF) signaling is considered as a central mechanism of body adapting to hypoxia. Hypoxia-inducible factor prolyl hydroxylase inhibitor FG-4592 (Roxadustat) is a first-in-class HIF stabilizer for the treatment of patients with renal anemia. The current study aimed to investigate whether FG-4592 could protect against ischemia/reperfusion (I/R)-induced kidney injury via inhibiting inflammation. Here, efficacy of FG-4592 was evaluated in a mice model of I/R-induced AKI. Interestingly, improved renal function and renal tubular injuries, combined with reduced kidney injury molecule-1 were observed in the mice with FG-4592 administration. Meanwhile, inflammation responses in FG-4592-treated mice were also strikingly attenuated, as evidenced by the decreased infiltration of macrophages and neutrophils and down-regulated expression of inflammatory cytokines. In vitro, FG-4592 treatment significantly protected the tubular epithelial cells against hypoxia-induced injury, with suppressed inflammation and cell injuries. In summary, FG-4592 treatment could protect against the I/R-induced kidney injury possibly through diminishing tubular cells injuries and suppression of sequence inflammatory responses. Thus, our findings definitely offered a clinical potential approach in treating AKI.

Viral metagenomics reveals sapoviruses of different genogroups in stool samples from children with acute gastroenteritis in Jiangsu, China.

Sapoviruses (SAVs), including several genogroups (GI to GV), are one of the causes of acute gastroenteritis (AGE). In this study, viral metagenomics revealed the presence of sapoviruses of different genogroups in stool from children with AGE. Eight different complete SAV genomes were determined, of which five belonged to GI and the other three belonged to GII, GIV and GV, respectively. Although they were highly similar to published sequences, the GIV and GV were the first complete genome sequences of these SAVs found in China. In a prevalence investigation, 19% of subjects with AGE were positive for SAVs, while none of the control group was positive.

Gender, Age, and Concomitant Diseases of Colorectal Diverticulum in China: A Study of 7,964 Cases.

AIM: To analyze the epidemiological features of colorectal diverticulum (CRD) in China. METHODS: We retrospectively analyzed CRD patients in 8 tertiary hospitals located in 5 regions of China from 2000 to 2016. The detection rates, number and distribution, demographic information, concomitant disorders, and their associations were investigated. RESULTS: Of 3,446,118 cases, 7,964 (2.3%) were CRD with a mean age of 56 years (11-92 years). The detection rate increased yearly and with increasing age. Males had a higher detection rate than females (3.0 vs. 1.47%, p < 0.01) and 1.8-times higher increase rate. The detection rate increased with age; however, females of > 60 years had a 2.8-times increasing rate than males. CRD occurred most frequently in the right-side colon, followed by rectum. Multiple diverticula were common in males and increased with age, with a 3-times higher increase rate than single lesion. Single-segment CRD occurred more frequently in males than in females (80.1 vs. 76.4%, p < 0.01). Concurred colon polyps were seen in 51.05% cases. CONCLUSION: CRD detection rates increased annually and with age, particularly in senior females in China. Multiple diverticula were common in males and increased with age. CRD was predominant in the right-side colon. Polyps are the most common comorbidity associated with CRD.

Aged men share the sperm protein PATE1 defect with young asthenozoospermia patients.

STUDY QUESTION: Does a defect in the human sperm-located protein prostate and testis expressed 1 (PATE1) exist in both aged men and young asthenozoospermia patients? SUMMARY ANSWER: A defect in sperm PATE1 exists in both aged men and young asthenozoospermia patients, and an antibody against PATE1 can decrease human sperm motility and zona-free hamster oocyte penetration. WHAT IS KNOWN ALREADY: Both aged men and young asthenozoospermia patients have poor sperm quality. The PATE1 protein seems to mediate sperm-egg interactions; however, the mechanisms are still unknown. STUDY DESIGN, SIZE, DURATION: This was a case-control study including 60 young fathers (aged 28-32 years) and 60 aged fathers (68-72 years old) who donated semen by masturbation after 7 days of sexual abstinence. Comparative sperm proteome analysis from the young fathers and aged fathers was performed to discover key proteins. The target protein PATE1 was chosen and validated by western blotting and immunohistochemistry. Quantitative assessment of sperm PATE1 protein was performed on sperm from 60 young fathers, 60 aged fathers and 110 young asthenozoospermia patients. Furthermore, an antibody against PATE1 assay was used to test whether PATE1 participated in sperm motility and penetration of zona-free hamster egg. PARTICIPANTS/MATERIALS, SETTING, METHODS: Samples were pooled and separated by two-dimensional gel electrophoresis followed by identification by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Western blotting and immunohistochemistry were used to validate the confidence of proteomic data. Sperm immunofluorescence quantification experiments disclosed whether the aged men indeed shared the same PATE1 defect with 110 young asthenozoospermia patients. The sperm motility test and penetration of zona-free hamster egg assay were performed for PATE1. MAIN RESULTS AND THE ROLE OF CHANCE: Twenty-two sperm proteins with significant differential expression between young adults and aged men were identified (P < 0.05, mean ratio >1.5), including 13 proteins with decreased expressions with aging. Based on bioinformatics, PATE1 was chosen for further study, and exhibited similar changes in expression level and localization on sperm from aged men and young asthenozoospermia patients. Antibody blocking revealed that PATE1 was involved in sperm-egg penetration and sperm motility. LIMITATIONS, REASONS FOR CAUTION: Before any clinical application of PATE1 as a biomarker for the diagnosis of male infertility, more cases should be used to evaluate confidence in this approach. WIDER IMPLICATIONS OF THE FINDINGS: This study revealed a common molecular basis underlying the decline in sperm quality in the natural aging process and in young men with asthenozoospermia. The data should greatly contribute to the development of molecular evaluation of sperm quality, and the diagnosis and treatment of asthenozoospermia. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grants from the National Natural Science Foundation of China (NO. 81300533, 81370013 and 81000277) and Shandong Provincial Natural Science Foundation, China (ZR2013HQ002, ZR2014HQ068). The authors declare no competing financial interests.

[The Optimal Storage Condition and Storage Time of Umbilical Cord Blood from Collection to Preparation].

OBJECTIVE: To explore the optimal storage condition and time of umbilical cord blood from collection to preparation. METHODS: Collect cord blood samples from 30 healthy newborns, with each new born's umbilical cord blood was divided into two parts on average. One part was stored in cold storage (4 ℃) and the other was stored at room temperature (20-24 ℃). Samples were taken at 24, 36, 48, 60 and 72 h, respectively, total nucleated cells (TNC) count and TNC viability was analyzed. Flow cytometry was used to detect the ratio of viable CD34+ cells to viable CD45+ cells and viability of CD34+ cells, and colony-forming unit-granulocyte-macrophage (CFU-GM) count was performed by hematopoietic progenitor cell colony culture. The change trend of each index over time was observed, and the differences in each index was compared between cold storage and room temperature storage under the same storage time. RESULTS: The TNC count (r 4 ℃=-0.9588， r 20-24 ℃=-0.9790), TNC viability (r 4 ℃=-0.9941， r 20-24 ℃=-0.9970), CD34+ cells viability (r 4 ℃=-0.9932， r 20-24 ℃=-0.9828) of cord blood stored in cold storage (4 ℃) and room temperature storage (20-24 ℃) showed a consistent downward trend with the prolongation of storage time. The percentage of viable CD34+ cells (r 4 ℃=0.9169， r 20-24 ℃=0.7470) and CFU-GM count (r 4 ℃=-0.2537， r 20-24 ℃=-0.8098) did not show consistent trends. When the storage time was the same, the TNC count, TNC viability, CD34+ cells viability and CFU-GM count of cord blood stored in cold storage were higher than those stored at room temperature. Under the same storage time (24, 36, 48, 60 or 72 h), TNC viability in room temperature storage was significantly lower than that in cold storage (P <0.001), but TNC count, percentage of viable CD34+ cells and CFU-GM count were not significantly different between room temperature storage and cold storage. When stored at room temperature for 24 h and 36 h, the viability of CD34+ cells was significantly lower than that in cold storage (P <0.001, P <0.01), when the storage time for 48, 60 and 72 h, there was no significant difference in the CD34+ cells viability between room temperature storage and cold storage. CONCLUSION: It is recommended that cord blood be stored in cold storage (4 ℃) from collection to preparation, and processed as soon as possible.

Therapeutic evaluation of rosuvastatin on lipids and endothelial cell functionalities in coronary artery lesions coinciding with hyperlipidemia.

OBJECTIVE: To analyze the changes of blood lipids and endothelial cell function in patients with coronary heart disease complicated with hyperlipidemia after treatment with rosuvastatin. METHODS: A total of 120 patients with coronary heart disease and hyperlipidemia diagnosed from December 2020 to December 2021 were retrospectively included. Depending on the differences of their treatment strategies, patients were divided into the study group (60 patients were treated with rosuvastatin using the conventional treatment) and the control group (60 patients were treated with the conventional treatment). Dynamic blood lipid level monitoring was performed on the two groups of patients. The changes of cardiac function and hemorheology indexes were evaluated before and after the treatment. Analyze the difference of vascular endothelial function index between the two groups before and after the treatment. Count the occurrence of adverse reactions during the intervention period of the two groups. RESULTS: Before the treatment, there was no significant difference between the two groups in total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C), left ventricular ejection fraction (LVEF), left ventricular end-systolic diameter (LVDS), left ventricular end-diastolic diameter (LVEDD), fibrinogen content, plasma viscosity, nitric oxide (NO), and endothelin (ET) levels (P>0.05). At 60 days of treatment, there was no significant difference between the two groups in TC, TG, LDL-C, LVDS, and LVEDD. The fibrinogen content, plasma viscosity, and ET level were lower than those in the control group (P<0.05). The HDL-C, LVEF, and NO levels were higher than those in the control group (P<0.05). There was no significant difference in the total incidence of adverse reactions between the two groups (8.33% vs 13.33%) (P>0.05). CONCLUSION: Resuvastatin can reduce the level of blood lipids in patients with coronary heart disease and hyperlipidemia and improve the hemorheology indexes and cardiac function of patients. Its mechanism may be related to the regulation of vascular endothelial cell function in patients with coronary heart disease.

Dose study of rosuvastatin calcium in the treatment of coronary heart disease and hyperlipidemia.

OBJECTIVE: To study the clinical efficacy of different doses of rosuvastatin to treat elderly patients with senile coronary heart disease and hyperlipidemia. METHODS: By means of retrospective analysis, 150 elderly patients with coronary heart disease and hyperlipidemia who were treated in Zhangjiakou First Hospital from January 2020 to December 2020 were selected as the study subjects. They were divided into three groups (50 patients in each group) according to the different treatment methods. All patients were given routine treatment for coronary heart disease and hyperlipidemia. At the same time, group A got 5 mg of rosuvastatin calcium per day, group B got 10 mg and group C got C, 20 mg. After 4 months of continuous treatment, changes of blood lipid level, inflammatory factors, and cardiac function in the three groups were compared before and after treatment. Finally, the incidence of adverse reactions in the three groups was statistically compared. RESULTS: After 4 months of treatment, the levels of TC, LDL, and TG in group B were significantly lower than those of group A, and the levels of HDL were significantly higher than those in group A (P<0.05). There was no significant difference of the above indicators between groups B and C after 4 months of treatment (P>0.05). Using 2 months, 3 months, and 4 months of therapy as time points, the blood lipid levels of the B and C groups was lower than in group A (P<0.05); Serum hs-CRP and TNF of patients in group B and group C after 4 months of treatment were significantly lower than those of group A (P<0.05); The LVEF comparison between groups showed that C was higher than A (P<0.05); The occurrence rate between adverse reactions during the 4 months of medication did not have statistical significance (P>0.05). CONCLUSION: Rosuvastatin calcium canimprove the clinical symptoms of elderly patients with coronary heart disease complicated by hyperlipidemia, and can improve the blood lipid level, cardiac function and the level of inflammatory factors in the body, but the clinical effect is not significantly improved by increasing the application dose. This suggests that the daily application dose should be 10 mg.

Comparing high-dose dual therapy with bismuth-containing quadruple therapy for the initial eradication of Helicobacter pylori infection on Hainan Island: A randomized, multicenter clinical trial.

BACKGROUND: Traditional bismuth-containing quadruple therapy, as a first-line eradication treatment for Helicobacter pylori (H. pylori), has several disadvantages, including drug side effects, low medication adherence, and high costs. Trials of high-dose dual treatment have demonstrated its advantages, which include good safety and adherence profiles. In this study, we investigated the efficacy, safety, and compliance of a high-dose dual therapy when compared with bismuth-based quadruple treatment for the initial eradication of H. pylori infection on Hainan Island, China. METHODS: We randomized 846 H. pylori-infected patients into two groups. A bismuth-containing quadruple therapy group was administered the following: esomeprazole 20 mg, amoxicillin 1000 mg, and clarithromycin 500 mg twice daily, and colloidal bismuth pectin in suspension 150 mg three times/day for 2 weeks. A high-dose dual therapy group was administered the following: esomeprazole 20 mg four times/day and amoxicillin 1000 mg three times/day for 2 weeks. Patients were given a 13C urea breath test at 4 weeks at treatment end. Adverse effects and compliance were evaluated at follow-up visits. RESULTS: Eradication rates in the high-dose dual therapy group were: 90.3% (381/422, 95% confidence interval [CI]: 87.1%-92.9%) in intention-to-treat (ITT) and 93.6% (381/407, 95% CI: 90.8%-95.8%) in per-protocol (PP) analyses. Eradication rates were 87.3% in ITT (370/424, 95% CI: 83.7%-90.3%) and 91.8% in PP analyses (370/403, 95% CI: 88.7%-94.3%) for quadruple therapy, with no statistical differences (P = 0.164 in ITT and P = 0.324 in PP analyses). Adverse effects were 13.5% (55/407) in the dual group and 17.4% (70/403) in the quadruple group (P = 0.129). Compliance was 92.4% (376/407) in the dual group and 86.6% (349/403) in the quadruple group (P = 0.007). CONCLUSIONS: High-dose dual therapy had high eradication rates comparable with bismuth-based quadruple treatment, with no differences in adverse effects, however higher adherence rates were recorded.

Geographical Pattern Evolution of Health Resources in China: Spatio-Temporal Dynamics and Spatial Mismatch.

(1) Background: The rational allocation of limited medical resources is the premise of safeguarding the public health. Especially since the outbreak of COVID-19, the evolution dynamics and spatial mismatch of medical resources have been a focal and frontier issue in academic discussions. (2) Methods: Based on the competitive state model and spatial mismatch index, this paper uses GIS and Geodetector spatial analysis methods and three typical indicators of hospitals, doctors, and beds to conduct an empirical study on the evolutionary characteristics and degree of mismatch in the geographic pattern of health resources in China from 2010 to 2020 (the data are from official publications issued by the National Bureau of statistics in China), in two dimensions of resource supply (economic carrying capacity) and demand (potential demand or need of residents). (3) Results: The spatial pattern of health resources at the provincial level in China has been firmly established for a long time, and the children and elderly population, health care government investment, and service industry added value are the key factors influencing the geographical distribution of health resources. The interaction between the different influence factors is dominated by bifactor enhancement, and about 30-40% of the factor pairs are in a nonlinear enhancement relationship. Hospital, doctor, and bed evolution trends and the magnitude and speed of their changes vary widely in spatial differentiation, but all are characterized by a high level of geographic agglomeration, heterogeneity, and gradient. Dynamic matching is the mainstream of development, while the geographical distribution of negative and positive mismatch shows strong spatial agglomeration and weak spatial autocorrelation. The cold and hot spots with evolution trend and space mismatch are highly clustered, shaping a center-periphery or gradient-varying spatial structure. (4) Conclusions: Despite the variability in the results of the analyses by different dimensions and indicators, the mismatch of health resources in China should not be ignored. According to the mismatch types and change trend, and following the geographic differentiation and spatial agglomeration patterns, this paper constructs a policy design framework of "regionalized governance-classified management", in line with the concept of spatial adaptation and spatial justice, in order to provide a decision making basis for the government to optimize the allocation of health resources and carry out health spatial planning.

Modulating gut microbiota as an anti-diabetic mechanism of berberine.

Berberine, one of the main constituents of a Chinese traditional herb used to treat bacterial diarrhea, has an effect of lowering glucose, which has been recently confirmed by many studies. However, the mechanism of berberine's antidiabetic effect has not yet been well explained. Recent evidence suggests that the gut microbiota composition is associated with obesity and type 2 diabetes, which are closely associated with a low-grade inflammatory state. The protective effect against diabetes of gut microbiota modulation with probiotics or antibiotics has been confirmed in recent observations. Berberine has significant antimicrobial activity against several microbes through inhibiting the assembly function of FtsZ and halting the bacteria cell division. Because berberine acts topically in the gastrointestinal tract and it is poorly absorbed, berberine might modulate gut microbiota without systemic anti-infective activity. Our hypothesis is that gut microbiota modulation may be one mechanism of the antidiabetic effect of berberine. Our hypothesis may provide a novel explanation for berberine's therapeutic effect in patients with diabetes mellitus.

Gender, age, and concomitant diseases of melanosis coli in China: a multicenter study of 6,090 cases.

BACKGROUNDS AND AIMS: Melanosis coli (MC) is a noninflammatory, benign, and reversible colonic disorder, but its detection rates in China are unclear. We therefore aimed to analyze the epidemiological characteristics of MC in China. METHODS: We assessed the detection rates, associated factors and concomitant diseases of MC in the patients who underwent colonoscopy at eight medical centers across five regions of China between January 2006 and October 2016. All data were procured from the electronic database established at each participating institutions. RESULTS: Among the 342,922 included cases, MC was detected in 6,090 cases (detection rate = 1.78%, 95% confidence interval, 1.73%-1.82%) at a mean age of 60 years. The detection rate gradually increased yearly, and along with the increasing age regardless of gender, while a rapid increase presented in the patients ≥60 years of age (0.58% for ≤25 years, 1.22% for 25-59 years, and 3.19% for ≥60 years). The detection rate was higher in females than in males; however, the rate of per-year increase was higher in males than in females at age of ≥60 years, which was 1.85-fold of that in females. Among cancer, polyp, inflammation, and diverticula, polyp was the most common concomitant disease of MC and identified in 41.72% of MC patients. CONCLUSIONS: MC detection rates were increased annually and elevated in older patients, particularly in male patients. Males in the elderly population of ≥60 years were most likely to have MC. Colonic polyp is the most common concomitant disease of MC.

Novel fluoroimmunoassays for detecting ochratoxin A using CdTe quantum dots.

Novel direct and indirect competitive fluorescence-linked immunosorbent assays (cFLISA and icFLISA) for detection of ochratoxin A (OTA) were described using CdTe quantum dots (QDs) as fluorescent label. CdTe QDs were successfully synthesized, which had an emission wavelength of 615 nm. The high purity monoclonal antibody against OTA was prepared through cell thawing and the octylic acid-ammonium sulfate method. The OTA MAbs were successfully coupled with CdTe QDs, and which also retained the original biological activity. The 50% inhibition values (IC50 ) of the cFLISA and icFLISA were 0.630 ng/mL, 0.234 ng/mL, the limits of detection (LOD) were 7.06 × 10-3 and 4.15 × 10-3  ng/mL, and detection ranges were 7.06 × 10-3 - 18.34 ng/mL and 4.15 × 10-3 - 4.88 ng/mL, in-order. The recoveries were 96.0-104.7% along with coefficients of variation (CVs) below 10%. The FLISA provided novel method for determination of OTA and the potential of MAb-CdTe QDs for the establishment of fluorescent immunochromatographic test strip.

[The effect of matrix metalloproteinase-9 in granulocyte colony stimulation factor-induced stem cell mobilization].

OBJECTIVE: To investigate the effects of matrix metalloproteinase-9 (MMP-9) on granulocyte colony stimulation factor (G-CSF)-induced hematopoietic stem/progenitor cell (HSPC) mobilization in healthy donors of hematopoietic stem cells. METHODS: Peripheral blood (PB) samples and bone marrow (BM) blood samples were collected from 12 healthy donors of hematopoietic stem cell before and 5 days after G-CSF-induced mobilization. CD34(+) cells were isolated and purified. ELISA was used to detect the protein expression of MMP-9 in the peripheral blood and BM blood of the healthy donors. The protein expression of MMP-9 in the BM blood was detected by ELISA and immunohistochemistry, and the stromal cell-derived factor-1 (SDF-1) level in the BM blood was detected by ELISA. The mRNA expression of MMP-9 in the BM blood samples was detected by RT-PCR. HT1080 cells rich in MMP-9 were cultured. CD34(+) cells were co-cultured with the supernatant of HT1080 cell culture fluid. CD34(+) cells cultured in Iscove's modified Dulbecco's medium were used as control group. Fluorescence-activated cell sorter was used to detect the CXCR4 expression on the surface of the CD34(+) cells. In the transwell experiment CD34(+) cells were divided into 4 groups: control group, o-phenanthroline (MMP-9 chemical inhibitor, MPI) group, HT1080 sup group, and HT1080 + MPI group to be co-cultured with buffer, o-phenanthroline, supernatant of culture fluid of HT1080 cells, or supernatant of culture fluid of HT1080 cells Flow cytometry was used to calculate the cell migration capacity. RESULTS: The MMP-9 level of BM and PB of the healthy donors 5 days after G-CSF mobilization were 278 ng/ml +/- 34 ng/ml and 392 ng/ml +/- 284 ng/ml respectively, both significantly higher than those before G-CSF mobilization (42 ng/ml +/- 17 ng/ml and 27 ng/ml +/- 12 ng/ml respectively (P < 0.01 and P < 0.05). Western blotting showed that the SDF-1 level in the supernatant 5 days after G-CSF mobilization was 5.9 ng/ml +/- 1.0 ng/ml, significantly lower than that before G-CSF mobilization (7.2 ng/ml +/- 0.7 ng/ml, P < 0.05). The CXCR4 levels of the CD34(+) cell from both PB and BM blood were up-regulated after co-culture with the supernatant of HT1080 cells (both P < 0.05). The migration capacity of CD34(+) cells cultured in the supernatant of HT1080 cells was increased significantly (P < 0.05), however, this effect could be inhibited by MIP (P < 0.05). The PB WBC numbers of the G-CSF group and G-CSF + MPI group were 14.9 x 10(6)/L +/- 4.3 x 10(6)/L and 12.3 x 10(6)/L +/- 1.2 x 10(6)/L respectively, the PB WBC numbers of the G-CSF + MPI group was significantly lower than that of the G-CSF group (P < 0.05), however, significantly higher than that of the negative control group (6.8 x 10(6)/L +/- 2.5 x 10(6)/L, P < 0.05). The CFU of the G-CSF group was (84 +/- 10) U/2 x 10(5) MNC, significantly higher than that of the G-CSF + MPI group, (69 +/- 3) U/2 x 10(5) MNC (P < 0.05). The BM MNC number of the G-CSF group was 12.7 x 10(6)/L +/- 0.7 x 10(6)/L, not significantly different from that of the G-CSF + MPI groups (13.1 x 10(6)/L +/- 1.3 x 10(6)/L; P > 0.05). CONCLUSION: MMP-9 probably facilitates HSPC mobilization by degrading SDF-1, up-regulating CXCR4 expression on the CD34(+) cells, and increasing the migration ability of CD34(+) cells.

Efficient inhibition of human B-cell lymphoma xenografts with an anti-CD20 x anti-CD3 bispecific diabody.

Bispecific antibodies have been exploited both as cancer immunodiagnostics and as cancer therapeutics, and have shown promise in several clinical trials in cancer imaging and therapy. A number of bispecific antibodies against B-cell markers have been shown to be effective in vitro in mediating tumor cell lysis and in vivo in inhibiting tumor growth in animal models. We have constructed a bispecific diabody from the variable genes encoding two hybridoma-derived monoclonal antibodies directed against human CD20 on B cells and CD3 on T cells. The anti-CD20 x anti-CD3 diabody was expressed in a single Escherichia coli host and purified by a one-step affinity chromatography. The bispecific diabody bound as efficiently to both CD20- and CD3-positive cells as the respective parental antibodies, and was capable of cross-linking CD20-positive tumor cells and human T lymphocytes as shown by cellular rosetting. The diabody effectively lysed human B-lymphoma cells in the presence of T-enriched human peripheral blood lymphocytes (PBL). Further, when combined with human PBL and interleukin-2, the diabody significantly prolonged the survival of nude mice inoculated with human B-lymphoma cells. Taken together, our results suggest that an anti-CD20 x anti-CD3 diabody may have significant clinical application in the treatment of human CD20-positive B-cell malignancies.

[Effect of serum from patients with Graves' disease on colony forming unit-granulocyte monocyte].

OBJECTIVE: To investigate the effects of serum component from patients with Graves's disease (GD) on the growth of colony forming unit-granulocyte monocyte (GM-CFU). METHODS: Monocytes were obtained from 11 normal persons and 11 GD patients with leukopenia and culture together with sera from 11 GD patients with leukopenia, 11 GD patients without leukopenia, and 11 normal controls for 10 days. Inverted microscopy was used to count the number of colony. RESULTS: Thyroid stimulating immunoglobulin (TSI) and serum from GD patients with leukopenia significantly inhibited the formation of GM-CFU (P < 0.01) while methimazole, thyroxine and serum from GD patients without leukopenia did not have any effect on the formation of GFU-GM (P > 0.05). CONCLUSION: TSI and serum from GD patients with leukopenia remarkably inhibit GM-CFU growth. Autoimmune abnormality may play an important role in the pathogenesis of leukopenia in patients with GD.

[The effects of the short-term ex vivo expansion on the adhesion characteristics and chemotactic function of expanded ex vivo CD34(+) cells].

OBJECTIVE: To study the effect of ex vivo expansion on the adhesion activities and chemotactic function of umbilical cord blood (UCB) hematopoietic stem and progenitor cells (HSPCs). METHODS: CD34(+) cells isolated from fresh UCB samples were cultured in serum-free and stroma-free culture system. After 7, 10 and 14 days' culture, CD34(+) cells were re-selected from the expanded products. Stromal cell- derived factor-1 (SDF-1) 100 ng/ml was added into the experimental CD34(+) cells and the absorbance at 570 nm of all groups was examined. 20 micro g/ml fibronectin (Fn) was added and the spontaneous adhesion between CD34(+) and FN was detected by MTT method. The homing-related functions including expression of homing-related adhesion molecules (CAM), adhesion activity and chemotactic function of the re-selected CD34(+) cells were evaluated and compared with those of the initial fresh CD34(+) cells. RESULTS: (1) The expression of CD49d, CD44, CD11a and CD49e on expanded CD34(+) cells increased or sustained the same levels as those of the fresh isolated UCB CD34(+) cells, while the expression of CD62L, CD54 and CD31 on expanded CD34(+) cells declined during the culture. (2) The spontaneous adhesion between CD34(+) and FN and SDF-1-induced adhesion continuously increased in the course of the first 10-day culture. The spontaneous adhesion rate and SDF-1-induced adhesion rate on day 0, day 7 and day 10 were 28% and 63%, 60% and 70%, 63% and 90% respectively. (3) The migration efficiency of re-selected CD34(+) cells on day 7 was almost the same compared to that of fresh CD34(+) cells. CONCLUSION: The expanded HSPCs sustain most of the homing-related characteristics and activities during one-week culture while extended culture may partly impair their intrinsic homing potential.

[The effects of cytokines mediated ex vivo expansion on the cell adhesion molecule expression of cord blood hematopoietic stem and progenitor cells].

OBJECTIVE: To compare the expression of cell adhesion molecules (CAMs) among VLA-4 (CD49 d), VLA-5 (CD49e), LFA-1 (CD11a), L-selectin (CD62L), and PECAM-1 (CD31) which are more related to the homing of hematopoietic stem and progenitor cells (HSPC) on the ex vivo expanded CD34+ subset with that of fresh isolated AC133+ cells. METHODS: AC133+ cells selected from fresh cord blood (CB) samples were cultured in QBSF-60 serum-free media in the presence of Flt-3 ligand + SCF + TPO (FST), with initial addition of IL-3 for up to 2 week. Expansion potential and the expression of above CAMs were evaluated at day 0, day 7, day 10 and day 14. RESULTS: (1) Simultaneously numerical expansion of various HSPC was constantly observed during the culture, and the fold expansion of AC133+ cells and CD34+ cells on day 14 were 33.50 and 64.56 respectively; (2) The number of CD34+ subsets expressing the above adhesions were all increased at different degrees (from 20 fold to 160 fold). (3) The expressions of CD11a, CD49d, and CD49e on ex vivo expanded CD34+ cells were increased as compared to their baseline levels, but the percentage of CD62L+ and CD31+ subpopulations in CD34+ cells were decreased. CONCLUSIONS: Our short-term culture system can not merely support the simultaneous expansion of CB derived AC133+ cells, but the expanded hematopoietic progenitors may well sustained the expression of homing-related adhesion molecules.

Intestinal microbiota and type 2 diabetes: from mechanism insights to therapeutic perspective.

The incidence of type 2 diabetes (T2DM) is rapidly increasing worldwide. However, the pathogenesis of T2DM has not yet been well explained. Recent evidence suggests that the intestinal microbiota composition is associated with obesity and T2DM. In this review, we provide an overview about the mechanisms underlying the role of intestinal microbiota in the pathogenesis of T2DM. There is clear evidence that the intestinal microbiota influences the host through its effect on body weight, bile acid metabolism, proinflammatory activity and insulin resistance, and modulation of gut hormones. Modulating gut microbiota with the use of probiotics, prebiotics, antibiotics, and fecal microbiota transplantation may have benefits for improvement in glucose metabolism and insulin resistance in the host. Further studies are required to increase our understanding of the complex interplay between intestinal microbiota and the host with T2DM. Further studies may be able to boost the development of new effective therapeutic approaches for T2DM.

Infusion of human umbilical cord­derived mesenchymal stem cells effectively relieves liver cirrhosis in DEN­induced rats.

Cirrhosis is the long­term outcome of chronic hepatic injury and no effective therapy is currently available for this disease. Mesenchymal stromal cells (MSCs) are multipotent cells that are easily acquired and amplified, and may be potential candidates for cell therapy against cirrhosis. This study aimed to determine the therapeutic effects of human umbilical cord­derived MSCs (hUCMSCs) for the treatment of liver cirrhosis and identify an effective method for engrafting MSCs. The model of liver cirrhosis was established by induction of diethylnitrosamine (DEN) in rats. The isolated hUCMSCs were identified by morphology, flow cytometry and multilineage differentiation; they were injected into the vein of DEN­induced rats at varied cell doses and infusion times. Biochemical analyses of the serum and histopathological analysis of the liver tissues were performed to evaluate the therapeutic effects of hUCMSCs in all treatment groups. The results indicated that isolated hUCMSCs were capable of self­replication and differentiated into multiple lineages, including osteoblast­, adipocyte­ and hepatocyte­like cells. Compared with the control group, administration of hUCMSCs at different cell doses and infusion times relieved DEN­induced cirrhosis to varying degrees. The therapeutic effects of hUCMSCs on liver cirrhosis gradually improved with increased cell dose and infusion times. The improvement of cirrhosis was due to the capacity of hUCMSCs to breakdown collagen fibers in the liver. It was demonstrated that infusion of hUCMSCs effectively relieved liver cirrhosis by facilitating the breakdown of collagen fibers in a dose­dependent manner and multiple infusions caused a relatively greater improvement in cirrhosis compared with a single infusion of hUCMSCs.