Network pharmacological analysis and experimental study of Pulsatilla chinensis against inflammatory injury caused by pneumonia in mice infected with influenza virus FM_1 / 中国中药杂志

Network pharmacology and the mouse model of viral pneumonia caused by influenza virus FM_1 were employed to explore the main active components and the mechanism of Pulsatilla chinensis against the inflammatory injury of influenza virus-induced pneumonia. The components and targets of P. chinensis were searched from TCMSP, and the targets associated with influenza virus-induced pneumonia were searched from GeneCards. The common targets between P. chinensis and influenza virus-induced pneumonia were identified with Venn diagram established in Venny 2.1. The herb-component-disease-target(H-C-D-T) network was constructed by Cytoscape 3.7.2. The above data were imported into STRING for PPI network analysis. Gene Ontology(GO) enrichment and KEGG pathway enrichment were performed with DAVID. BALB/cAnN mice were infected with the influenza virus FM_1 by nasal drip to gene-rate the mouse model of pneumonia. Immunohistochemistry was adopted to the expression profiling of inflammatory cytokines in the lung tissues of mice in the blank group, model group, and P. chinensis group 1, 3, 5, and 7 days after infection. The pathological changes of lung and trachea of mice in blank group, model group, and P. chinensis group were observed with light microscope and scanning electron microscope at all the time points. The network pharmacological analysis indicated that 9 compounds of P. chinensis were screened out, with a total of 57 targets, 22 of which were overlapped with those of influenza virus-induced pneumonia. A total of 112 GO terms(P<0.05) were enriched, including 81 terms of biological processes, 11 terms of cell components, and 20 terms of molecular functions. A total of 53 KEGG signaling pathways(P<0.05) were enriched, including TNF signaling pathway, influenza A signaling pathway, NF-κB signaling pathway, MAPK signaling pathway and other signaling pathways related to influenza/inflammation. In the P. chinensis group, the expression of TNF-α and IL-1 in the lung tissue was down-regulated on the 3 rd day after infection, and that of IL-6 in the lung tissue was down-regulated on the 5 th day after infection. Light microscopy and scanning electron microscopy showed that P. chinensis significantly alleviated the pathological damage of lung and trachea compared with the model group. This study reflects the multi-components, multi-targets, and multi-pathways of P. chinensis against influenza virus-induced pneumonia. P. chinensis may reduce the production of proinflammatory cytokines and mediators and block the pro-inflammatory signaling pathways to alleviate viral pneumonia, which provides reference for future research.

Tetramethylpyrazine inhibits PM2.5-induced vascular smooth muscle cell proliferation by down-regulating JNK phosphorylation / 中国药理学与毒理学杂志

OBJECTIVE To investigate the inhibitory effect and the possible mechanism of tetra-methylpyrazine (TMP) in preventing vascular smooth muscle cells (VSMCs) proliferation induced by fine particulate matter (PM2.5). METHODS PM2.520, 200 and 400 mg · L-1 was added to VSMCs for 24 h, the survival of VSMCs was measured by MTT assay, the protein levels of p-c-Jun N-terminal kinase (JNK) and fibroblast growth factor receptor-1 (FGFR-1) in the VSMCs were detected by Western blotting, while the levels of vascular cell adhesion molecule-1 (VCAM-1), endothelin-1 (ET-1) and nitric oxide (NO) in the VSMCs were analyzed by ELISA, radioimmunoassay and nitrate reductase method, respec-tively. TMP 20, 200 and 2000 mg·L-1 or a specific inhibitor of JNK SP60012510μmol·L-1 was added into the VSMCs to observe the effect of TMP. RESULTS Compared with the normal control group, PM2.5200 and 400 mg·L-1 significantly increased the A570 nm vaule, the protein levels of p-JNK and FGFR-1,the levels of VCAM-1 and ET-1, but decreased the level of NO (P<0.01), while there were no significant changes in PM2.520 mg·L-1 group. Compared with the PM2.5200 mg·L-1 group, TMP 200 and 2000 mg·L-1 pre-treatment markedly decreased the A570 nm vaule, the protein levels of p-JNK and FGFR-1, the levels of VCAM-1 and ET-1, but increased the level of NO (P<0.01), while there were no significant changes in TMP 20 mg · L-1 pre-treated group. Moreover, the effects of TMP were significantly enhanced by the co-incubation of TMP 2000 mg · L-1 with SP60012510 μmol · L-1, compared to the TMP 2000 mg · L-1 pre-treated group (P<0.05, P<0.01). CONCLUSION TMP displays a significant inhibitory effect against VSMC proliferation induced by PM2.5. The mechanism may be related to the inhibition of JNK phosphor-ylation, and the regulation of FGFR-1 protein expression and VCAM-1, ET-1 and NO levels.

Treating ISR after PCI by LIU Zhong-yong from Turbidity Syndrome / 中国中医药信息杂志

It is a thorny problem of modern medicine that the in-stent restenosis (ISR) after percutaneous coronary intervention (PCI). Combining with the etiology and pathogenesis of TCM, Professor LIU Zhong-yong believes that the root cause of ISR after PCI is the deficiency syndrome: menstruation gradually dying up, the heart yang qi deficiency; and the direct cause is excess syndrome: endogenous turbidity syndrome, heart vessel blockage. The cause for the formation of turbidity syndrome is cold, phlegm, blood stasis, poison, and dampness. Depending on the clinical manifestations, five kinds of syndromes were divided: cold turbidity stagnation, phlegm turbidity resistance, blood stasis blockage veins, poison turbidity, and dampness turbidity resistance. Professor LIU Zhong-yong also proposed relevant treatment for both symptoms and root causes, which provided new ideas and experience in the integrated TCM and Western medicine for ISR after PCI.

Clinical application of combined detection of tumor molecular markers in lung cancer / 肿瘤研究与临床

Objective To explore the values of combined detection of carcino embryonic antigen (CEA),carcinoma antigen 125 (CA 125),cytokerain 19 fragment 21-1 (CYFRA2 1-1),carcinoma antigen Scc-Ag (Scc-Ag) and neuron specific enolase (NSE) for diagnosis and treatment of lung cancer.Methods The concentrations of CEA,CA125,CYFRA21-1,Scc-Ag and NSE in 198 patients with lung cancer (lung cancer group),50 with benign pulmonary disease (benign disease group) and 50 healthies (health control group) were detected by electrochemiluminescence immunoassay (ECLIA).The concentration of Pro-GRP31-98 was detected by enzyme linked immunoassay.Results Six tumor markers in lung cancer group showed significant diffcrence compared with those of benign disease group and health control group (P ＜ 0.01).The concentrations of serum tumor molecular markers were significantly higher in Ⅲ,Ⅳ stage group than those in [,Ⅱ stage group.Before the treatment,the concentrations of serum tumor molecular markers in lymph node metastasis group and lymph node recurrence group were significantly higher than those in no lymph node metastasis group and no lymph node recurrence group after the treatment (P ＜ 0.01).The positive rate of CEA,CA125 in lung adenocarcinoma group,CYFRA21-1 and Scc-Ag in lung squamous cell carcinomas group and NSE and Pro-GRP31-98 in small cell lung carcinoma group were significantly different from the other two kinds of pathological types (P ＜ 0.01).The positive rates of CEA and CA125 combinated detection in lung adenocareinoma,CYFRA21-1 and Scc-Ag combinated detection in lung squamous cell carcinomas,NSE and Pro-GRP31-98 combinated detection in small cell lung carcinoma was significantly higher than the individual detection respectively.The sensitivitv rates of CEA,CA125,CYFRA21-1,Scc-Ag,NSE and Pro-GRP31-98 were 52.52 ％,51.01％,48.99 ％,53.03 ％,49.49 ％ and 50.50 ％,respectively.And the combinated detection sensitivity rate of the six tumor markers was 96.46 ％,which significantly higher than that of a single marker (P ＜ 0.01).Conelusions The tumor molecular markers of CEA,CA125,CYFRA21-1,Scc-Ag,NSE and Pro-GRP31-98 may be the indicators in diagnosis of lung cancer.The combined detection of serum tumor molecular markers would significantly improve the sensitivity of diagnosis for lung cancers.At the same time,it should be valuablc for determination of clinical stage,histological type and follow-up studies of lung cancer.

Research and clinical application on combined detection of serum tumor markers in early diagnosis of primary hepatocellular carcinoma / 中国医师进修杂志

Objective To investigate the value on clinical application of dynamic and combined detection of serum tumor markers alpha-fetoprotein (AFP),Golgi protein 73 (GP73),vascular endothelial growth factor (VEGF) and alpha-L-fucosidase (AFU) in early diagnosis and monitoring treatment of primary hepatocellular carcinoma.Methods Serum levels of AFP,GP73,VEGF,AFU in 98 patients with primary hepatocellular carcinoma (the malignant group) before and 3 months after treatment were detected by electrochemiluminescence immunoassay and enzyme-linked immunosorbent assay,which compared with the levels of 50 liver benign lesions (the benign group) and 50 healthy subjects (the normal group).Results The levels of serum AFP,GP73,VEGF,AFU in the malignant group before treatment were (219.16 ± 56.89) μg/L,(355.42 ± 109.26) μg/L,(88.21 ±24.22) μg/L,(276.51 ±83.20) U/L.These indexes in the benign group were (14.95 ± 3.26) μ g/L,(56.43 ± 15.72) μ g/L,(2.68 ± 1.27) μ g/L,(25.38 ± 11.17)U/L.These indexes in the normal group were (14.67 ± 3.07) μ g/L,(55.06 ± 14.21) μ g/L,(2.36 ± 1.14) μ g/L,(24.29 ± 10.10) U/L.There were statistical differences between the malignant group and the benign group,normal group (P ＜ 0.01).The levels of these indexes after treatment in the malignant group were (25.66 ±8.17) μg/L,(65.32 ±24.15) μg/L,(4.67 ± 1.89) μg/L,(35.23 ± 12.36) U/L,there were statistical differences between before treatment and after treatment (P ＜ 0.01).The sensitivity and accuracy of combined detection of these tumor markers in diagnosis of primary hepatocellular carcinoma were 95.92％ (94/98) and 93.24％(138/148),which were statistically higher than single detection(P＜ 0.05).Conclusion The dynamic and combined detection of serum tumor markers could be applied as supplementary means in early diagnosis and monitoring treatment of primary hepatocellular carcinoma.

The value of dynamic combined detection of serum tumor biomarkers in diagnosing primary hepatic carcinoma / 肿瘤研究与临床

Objective To investigate the value of combined detection of serum tumor biomarkers including alpha-fetoprotein (AFP),tumor specific growth factor (TSGF),golgi protein 73 (GP73) and osteopontin (OPN) in diagnosis of primmy hepatic carcinoma.Methods AFP,TSGF and GP73 levels were measured by electro-chemiluminescence immunoassay,and OPN levels by enzyme-linked immunosorbent assay (ELISA) in 122 cases of primary hepatic carcinoma,50 cases of liver benign lesions and 50 cases of healthy control.The biological parameters and the levels of AFP,TSGF,GP73 and OPN were studied.Results The serum levels of AFP,TSGF,GP73 and OPN in primary hepatic carcinoma were higher than those in the liver benign disease group and the normal control group (all P ＜ 0.01).Sex and age of the patients at diagnosis showed no significant association with the levels of the four serum tumor biomarkers in the primary hepatic carcinoma groups (all P ＞ 0.05).But the tumor size,amount,tumor stage,metastasis and recurrence showed significant association with the levels of those in the primary hepatic carcinoma groups (all P ＜ 0.05).The sensitivity,specificity and veracity of serum AFP,TSGF,GP73 and OPN as individual diagnostic marker was only 57.38 ％,68.85 ％,70.49 ％ and 69.67 ％,respectively.The sensitivity of combined detection of AFP and TSGF was 80.33 ％,and that of combined detection of AFP,TSGF and GP73 was 85.25 ％.While,the sensitivity of the four serum tumor markers in combination was 98.36 ％,the accuracy was 95.65 ％.The sensitivity and accuracy of combined detection with the four serum tumor markers were significantly higher than those of the individual markers and other combination detection methods (all P ＜ 0.05).Conclusions Serum markers of AFP,TSGF,GP73 and OPN can serve as a means for diagnosis of primary hepatic carcinoma.Combined detection of the four serum tumor biomarkers can improve the sensitivity,accuracy and the negative predictive value,which is benefit to early diagnosis and interference.

Mechanism of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture against influenza immune inflammatory injury / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on mRNA expression of lung inflammatory cytokines and pulmonary pathological injury of mice infected by influenza virus, in order to discuss the mechanism of tonifying Qi traditional Chinese medicines against pulmonary immune inflammatory injury of infected mice.</p><p><b>METHOD</b>In different time phases after mice were infected with influenza virus FM1, the RT-PCR method was adopted to observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on five inflammatory cytokines TNF-α, IL-1, IL-6, IL-10 and IFN-γ, and the changes in pulmonary pathological injury of mice with viral pneumonia after intervention with tonifying qi traditional Chinese medicines.</p><p><b>RESULT</b>(1) Tonifying Qi traditional Chinese medicines significantly reduced the mRNA expression of TNF-α at 1-5 d and IL-1 mRNA expression at 7 d, may increase IL-1 mRNA expression in mouse lung at 3 d, significantly reduced IL-6 mRNA expression in mouse lung and increased IL-10 mRNA expression at 3-7 d, and significantly increased IFN-γ mRNA expression at 1 d. (2) Tonifying Qi traditional Chinese medicines could significantly inhibited and repaired pulmonary immune inflammatory injury of mice infected by FM1, which was most remarkable at 3-7 d after the infection with influenza virus FM1.</p><p><b>CONCLUSION</b>Tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture could resist pulmonary immune inflammatory injury and repair inflammatory injury by regulating the mRNA expression of imbalance inflammatory cytokines of organisms infected with influenza virus.</p>

The influence of representative herbs of clearing and detoxifying drugs effect on inflammatory cytokines expression of mice lung homogenate infected by influenza virus FM1 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In order to screen out a certain kind of traditional medicine which has a better role in immune regulatory, the influence of representatives of heat clearing and detoxicating herb on inflammatory cytokines protein expression of mice lung homogenate infected by FM1 have been observed.</p><p><b>METHOD</b>Modeling mice infected by FM1. On the first, third, fifth and seventh day after FM1 infection, tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-10 (IL-10), and gamma-interferon (IFN-gamma) expression in mice lung homogenate of normal control group, model control group, scutellari group, isatidis group, pulsatilla group, polygonum cuspidatum group and oldenlandia group have been tested by ELISA method.</p><p><b>RESULT</b>The expression of TNF-alpha, IL-6, IFN-gamma and IL-10 in mice lung homogenate reaches its peak on the third day after FM1 infection, significantly higher than the control group (P < 0.05). Scutellari and isatidis are two representatives of heat clearing and detoxicating herb, which can decrease the expression of TNF-alpha, IL-6 and IL-1 and increase the expression of IL-10, IFN-gamma. The effect are more pronounced and statistically significant (P < 0.05) on the third and fifth day after infection, pulsatilla, polygonum cuspidatum and oldenlandia can also regulate the inflammatory cytokines, but the effect are not so obvious as scutellari and isatidis.</p><p><b>CONCLUSION</b>Scutellari and isatidis, two representatives of heat clearing and detoxicating herb, have a good intervention on immune damage caused by influenza virus through adjusting the balance of inflammatory cytokines and anti-inflammatory cytokines.</p>

Study on the effect of realgar nanoparticles anti-adenovirus in vitro / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was to establish a model that adenovirus type 3 (HAdV-3) infected on Hep-2 cell in order to explore anti-adenovirus3 (HAdV-3) effect of Chinese medicine realgar in vitro.</p><p><b>METHOD</b>Use high-energy ball milling with distilled water to prepare realgar nanoparticles. The concentration of nanometer realgar was tested by molybdenum blue staining method and realgar nanoparticles' particle size was tested on Nano Series. The technique of cell culture with ribavirin as positiv control was to observe anti-adenovirus effect through prevention, treatment and direct inactivation of three kinds of drug delivery.</p><p><b>RESULT</b>This drug was found to be a potential inhibitor of HAdV-3 in a concentration-dependent manner with the median toxic concentration (TC50) of 0.649 microg/ml in Hep-2 Cell culture. The median inhibition concentration (IC50) was 0.255 microg/ml when drug was added before infection. The IC50 was 0.142 microg/ml when drug was added after virus infection, and it was 0.117 microg/ml as the drug was added after it mixed with virus. The therapeutic index (TI) was 2.55, 4.57 and 5.55 respectively.</p><p><b>CONCLUSION</b>The direct inactivation effect of realgar nanoparticles is the most evident in three drug deliveries manner with the same concentration in vitro.</p>

Polymorphism of killer cell immunoglobulin-like receptor gene and its correlation with leukemia / 中国实验血液学杂志

The study was purposed to investigate the polymorphism of killer cell immunoglobulin-like receptor (KIR) gene of the patients with leukemia and to explore the correlation between the KIR gene and susceptibility of leukemia. The KIR genotype of 50 patients with leukemia and 60 healthy controls in northern. Hans were analyzed by PCR-SSP. The results indicated that the present known 18 KIR genes were detected and identified. The frequencies of KIR 3DL3, 3DL2 and 2DL4 were 100% in all subjects, with the most frequent genotype KIR 3DP1 (0.86) followed by 2DP1, 2DL3, 3DL1, 2DL1, 3DS1, 2DL5, 2DS4, 2DS2, 1D, 2DS5, 2DL2, 2DS1, 2DS3 and 3DP1v in leukemia successively. Compared with the control, the KIR 3DL1 (0.60) and 2DL1 (0.57) were significantly lower in the leukemia patient group than that in the control group (1.00) (P < 0.01). It is concluded that the polymorphism of KIR gene is associated with susceptibility of leukemia in Hans. There may be a negative correlation between pathogenesis of leukemia and KIR 3DL1, KIR 3DS1, KIR 2DL1, KIR 2DL5 genes.

The dynamic effect of yiqi qingwen jiedu heji on the protein expressions of cytokine IFN-gamma, TNF-alpha, IL-10 and IL-6 in the lungs of mice infected by IV FM1 / 中国中药杂志

<p><b>OBJECTIVE</b>To find the function and functioning mechanism of Yiqi Qingwen Jiedu Heji in resisting influenza immune damage by studying its effect on the expressions of cytokine.</p><p><b>METHOD</b>Taking IV FM1 infected mice as its model and doing ELISA (double antibody sandwichenzyme linked immunosorbent assay), we dynamically observed the change of cytokine TNF-alpha, IL-6, IFN-gamma and IL-10 after giving Yiqi Qingwen Jiedu Heji treatment.</p><p><b>RESULT AND CONCLUSION</b>After the mice are infected by influenza virus, their protein expressions of the model group are higher than those of the control group, of which TNF-alpha, IL-6, IFN-gamma reach the peak in three days. The three expressions of Yiqi Qingwen Jiedu Heji treated group are decreased and the decrease becomes remarkable on the third day, compared with those of the model group. However, the expression of IL-10 of the treated group is remarkably increased. It indicates that Yiqi Qingwen Jiedu Heji can resist the expressions of TNF-alpha, IL-6 and IFN-gamma pro-inflammatory cytokine,increase the expression of IL-10, and thus, alleviate inflammatory injury. So the clinical application of such medicine can shorten the course of disease.</p>

Prediction of acute GVHD after umbilical cord blood transplantation by HLA three-dimensional structure matching / 中国实验血液学杂志

To explore a method of predicting acute graft versus host disease (aGVHD) after unrelated cord blood transplantation (UCBT), the HLA-A, -B, -DRB1 molecular three-dimensional structures in 25 patients with blood disorder who underwent UCBT and their donors were modeled by using molecular modeling technique. First, full amino acid sequences of each HLA antigen from HLA data banks were loaded down, and then amino acid sequence of extracellular antigen binding region was chosen. Third step, SPDBV software of SWISS-MODEL server was used to modeling the three-dimensional structures of each different allele of HLA-A, -B and -DRB1 between patients and donors and the parameter "root mean square deviation" (RMSD) was used to indicate the structure differences. Last, RMSD of each different HLA allele of each donor-patient pair were added together to get total RMSD. The 25 patients were divided into 3 groups: the first group did not develop aGVHD; the second group developed aGVHD graded I-II and the third group developed aGVHD graded III-IV. The results showed that in the 25 patients divided into three groups, 8 patients in the first group did not develop aGVHD (32%); 13 patients in the second group developed grade I-II of aGVHD (52%) and 4 patients in the third group developed aGVHD III-IV (16%). The total RMSDs of each group were 0.24 +/- 0.15, 0.25 +/- 0.14 and 0.47 +/- 0.22 respectively. The total RMSD of the third group was significantly higher than that of the other two groups. In conclusion, utilization of modeling HLA molecular three-dimension can predict the severe aGVHD after UCBT quickly, simply and accurately. It provides scientific basis in choosing a optimal cord blood donor to avoid severe aGVHD for physicians and the cord blood banks. And it is instructive too to direct the application of immunosuppressive agents after transplantation in clinic.

Distribution features of HLA-DRB1 allele frequencies in normal cord blood donors of 3 438 Shandong Hans / 中国实验血液学杂志

The objective of the study was to research the distribution features of HLA-DRB1 alleles in Shandong Hans' population and explore the possibility of finding the cord blood donor of HLA-DR matched to perform the stem cell transplantation for more patients from larger region of China and even other areas in the world. The subjects of the study were drawn from 3 438 Shandong Hans donors in Shandong Umbilical Cord Blood Bank and were tested by PCR-SSP technique for HLA-DR low resolution typing. The result indicated that the most prevalent five alleles of HLA-DRB1 locus were DRB1 * 15 (0.1817), * 07 (0.1369), * 09 (0.1221), * 04 (0.1084) and * 12 (0.1038). The DR18 has the lowest gene frequency 0.0003, while DRB1 * 10, * 16 and * 01 showed lower gene frequencies (GF), which GF were 0.0151, 0.0262, and 0.0322 respectively. As compared the HLA-DRB1 GF of Shandong Hans with those of other Han Chinese and other ethnic populations, there were unique distributed features of DRB1 alleles among various races populations, and those among the studied population groups from various regions with the same race origin. The difference from various regions in the same race was less than that among different races. In conclusion, a patient of Han Chinese is easier to search a DR-matched cord blood donor in Shandong Umbilical Cord Blood Bank, especially from northern Hans. No DRB1 allele is unique to single racial group and majority of DRB1 low-resolution phenotypes are common to all studied groups. It is reasonable for some patients from other races including Caucasian and Japanese to receive a transplant of cord blood stem cell matched with HLA-DR in Shandong Umbilical Cord Blood Bank.

Statistical analysis of 4000 umbilical cord blood units preserved in shandong cord blood bank / 中国实验血液学杂志

The current study analyzed the data of 4 000 umbilical cord blood (UCB) units collected in Shandong Cord Blood Bank from the end of 1999 to March 2001. The averages of nucleated cells and CD34(+) cells were more than 1.2 x 10(9) and 3.9 x 10(6) per UCB unit respectively, and more than 1.5 x 10(9) nucleated cells per UCB unit were obtained in 768 UCB units. These UCB units are suitable for transplantation in patients with a body weight greater than 40 kg. The analysis of HLA gene frequency showed that A2, A24, A11, B13, B51, DR15, DR7 and DR9 are the common halotypes in Shandong population and similar to those in the other areas of China. 40% patients could search out at least 1 UCB unit with 1 mismatched HLA locus in Shandong Cord Blood Bank.

Rich hematopoietic stem/progenitor cells in the human placenta tissue and placenta blood / 中国实验血液学杂志

Clinical transplantation evidence has indication that umbilical cord blood (UCB) can be useful in the hematopoietic reconstitution in the children, but not well in the adult patients because of the low cell count. The purpose of our study was to evaluate a new method for collection of blood cells from human placenta and umbilical cord. We have simultaneously harvested blood cells from umbilical cord (UCB), placenta blood (UPB) and placenta tissue (UPT) for their content of nucleated cells, CD34 (hematopoietic stem progenitor marker) positive cells. Result showed that the nuclear cell (NC) from UPB and UPT has three to four times than that from umbilical cord blood only, (8.3 +/- 1.04) x 10(8) (UCB), (16.33 +/- 5.54) x 10(8) (UPB), and (8.01 +/- 2.64) x 10(8) (UPT). CD34(+) cells are (0.77 +/- 0.01) x 10(6), (1.25 +/- 0.55) x 10(6) and (4.21 +/- 1.90) x 10(6) respectively. The cells from UPB and UPT have more survival ability than the cells from UCB in the long-term cell culture condition. It is clear that the blood stored in the liquid nitrogen did not show large loss of total nucleated cell count and CD34(+) cells. It was observed that UPT and UPB contained more suppressor lymphocytes, which may be important in prevention of graft-versus-host disease. In conclusion, our data may have implications for the development of placental blood collection together with umbilical cord blood banking for the stem cell transplantation.

Operating Procedure of Collection, Processing and Preservation of 3000 Units Umbilical Cord Blood in Shangdong Cord Blood Bank / 中国实验血液学杂志

The experience with the umbilical cord blood (UCB) stem cells for unrelated transplantation from our 3 000 UCB storage was described. UCB, collected from closed blood bags, were mixed with hydroxyethyl starch for nucleated cell (NC) enrichment. After finishing CD34 analysis, culture of hematopoietic progenitors (CFU-GM and CFU-GEMM) assays, microbial culture, HLA Class I (A, B) serology and class II (DR) low resolution SSP typing, cord blood units are stored in the liquid nitrogen for clinical applicatoin. Cord blood contained an average of nuclear cell (NC) (1.2 +/- 0.6) x 10(9), CD34(+) cells (3.0 +/- 3.7) x 10(6), CFU-GM (1.1 +/- 0.7) x 10(6) and CFU-GEMM (1.1 +/- 1.2) x 10(6) for storage and the recovery rates were 91%, 88%, 85% and 82%, respectively. The recovery rates for red blood cell and Hb were (39 +/- 9)% and (40 +/- 8)%, respectively. The storage volume was (35.1 +/- 7.1) ml in a 50 ml storage bags. The mean time from collection to processing of 15 hours (range 4 - 24 hours) had no influence on cell viability. The cell viability before processing is more than 95% and 92% after UCB thawing. The recovery rates of NC, CD34(+) cells and CFU-GM post-thawing were 96%, 90% and 91%, respectively. There were no HIV antibody (HIVAb) positive in all of UCB units. For an incidence of processed samples, infection with syphilis, HBsAg, HBcAb, HCVAb, CMV, bacterial contamination and abnormal hemoglobin were 0.1%, 0.8%, 3.2%, 0.2%, 87.1%, 1.2% and 0.1%, respectively. More than 3 HLA loci matched can be found for random patients in our cord blood bank and 6 HLA loci matched have 5%. For transplantation with nucleated cell counts of > 2.7 x 10(7) cells/kg, our cord blood bank will be able to provide all of the umbilical cord blood stem cell samples for children and 50% of units can be used for some of adult recipients transplantation in the country. It is concluded that: (1) The large cord blood banking for 20 000 UCB storage is feasible in China. (2) Our system of whole procedure and methods is functionable for supplying qualified cord blood units in transplantation. (3) The volume for collection is critical to the yield of CD34(+) cells or hematopoietic progenitor cells, however cord blood NC is also important and proportional with CD34(+) cells. Only the units containing more than 8 x 10(8) cells and more than 60 ml of cord blood can be in the procession for storage.