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Objective: To understand the epidemiologic characteristics of endemic typhus in Baoshan city., Methods: Epidemiological data were collected and characteristics were analyzed. IgG antibody (Ab) of Rickettsia mooseri and Orientia tsutsugamushi in serum of patients were tested using both Weil-Felix and IFA method. The Rickettsia mooseri gltA gene, Rickettsia prowazekii gltA gene, Orientia tsutsugamushi 56 kDa protein gene, SFGR ompA gene, Ehrlichia sp. 16S rRNA gene and Anaplasma sp. 16S rRNA gene in spleen of mice were examined by PCR., Results: Fifty-eight endemic typhus cases were found in Longyang district of Baoshan city, during July to August, 2009. Among them, 48 cases were confirmed by clinical diagnosis and 10 cases by laboratory tests. The Ab of Orientia tsutsugamushi Karp serotype was detected in 3 cases from laboratory diagnosis. The spleen samples from 85 Rattus flavipectus were tested using PCR. Of them, 3 samples for Rickettsia mooseri gltA gene showed positive (positive rate was 3.5%), and the homology of 3 Rickettsia mooseri and Rickettsia mooseri Wilmington strain (GenBank U59714.1) was 100% through comparing gene sequence. The results of PCR for detecting Rickettsia prowazekii, Orientia tsutsugamushi, SFGR, Anaplasma sp. and Ehrlichia. sp were all negative., Conclusion: The outbreak of endemic typhus was confirmed in Longyang district of Baoshan city through epidemiological data, clinical diagnosis and laboratory tests. Rickettsia mooseri DNA was detected in the dominant Rattus flavipectus, suggesting that endemic typhus did exist in the local areas.

The aim of this study was to detect DNA damage during expansion ex vivo of umbilical cord blood (UCB) hematopoietic cells and explore the optimal harvest time for culture of CB hematopoietic cells. Mononuclear cells (MNCs) separated from UCB were cultured in a serum-free system supplemented with cytokines and colony forming units were assessed by semisolid culture at the same time. On day 0, 7, 14 and 21 cells were collected for single cell gel electrophoresis (SCGE) analysis and CFUs were also assayed by SCGE, CD34+ cells and CD133+ cells were quantitated by fluorescence-activated cell sorting (FACS). The results showed that the percentage of CD34+ and CD133+ cells was found to be highest after short-term culture (<14 days) and the cord blood DNA damage rate was observed to be less than 5.0% at earlier time points, but at day 21 the DNA damage rate was 28.2%, which was higher than that at day 0 (p=0.000), the tail length of the DNA comet was longer than that at day 0 (p=0.000). The tail lengths of DNA damage on other time points were not significantly different from that at day 0. It is concluded that the DNA damage rate is less than 5.0% after short-term (<14 days) culture of UCB cells ex vivo by using this method. After 14 days DNA damage rate increases significantly. The optimal harvest time of cord blood cells after culture ex vivo would be within 14 days.

Objective: To assess whether preservative-free 1% lidocaine is capable of destroying the LECs in age-related cataract (ARC) in order to provide scientific basis for pursuing safe and effective drugs to eliminate LECs in cataract surgery., Methods: Lens anterior capsule (LAC) specimens were collected from 75 patients (82 eyes) with age-related cataract (ARC), including forty males (44 eyes) and thirty-five females (38 eyes). The age range was 41 - 85 years, the mean age was 67.97 years. There were 34 cortical cataracts, 22 nuclear cataracts and 26 subcapsular cataracts. Capsule specimens were divided into 4 groups: balanced salt solution (BSS) group I and group II (exposed to BSS for 1 minute), lidocaine group (exposed to preservative-free 1% lidocaine for 1 minute) and the control group. Specimens were stained with trypan blue and alizarin red. Photomicrographs of each capsule were taken to observe the viability of LECs and to count the number of necrosis LECs. The pathologic changes of LECs were evaluated by histological methods (11 LAC, 22 pieces) as well as transmission and scanning electron microscopes (5 LAC, 10 pieces). In the control and BSS group I (23 LAC), one half of each capsule specimen was used for the control group and the other half was used for BSS group I. In lidocaine group and BSS group II (43 LAC), one half of each capsule specimen was used for lidocaine group and the other half was used for BSS group II., Results: The rate of necrosis LECs of the anterior capsules in the control group and BSS group I was (56.19 +/- 2.71)% and (57.23 +/- 1.98)%, respectively. The rate of necrosis LECs of the capsules in lidocaine group and BSS group II was (99.86 +/- 8.22)% and (57.64 +/- 7.00)%, respectively. Matching t-test showed that the rate of necrosis LECs in lidocaine group was greater than that in the BSS group II (t = 27.6781, P = 0.0000). There was no significant difference in the number of necrosis LECs between the control group and BSS group I (t = 2.0693, P = 0.0505). There was also no significant difference between males and females; between different cataract, types and between varying age groups (P > 0.05). After irrigated with lidocaine, LECs showed vacuoles and detached from the capsule, and many cavities appeared between the LECs and the capsule. The capsules of BSS and control group showed a normal layer of LECs attached to the capsule. Under transmission and scanning electron microscopes, in lidocaine group, the junction between the LECs and between the cells and the capsule were destroyed; many cells detached from the capsule and the rest arranged loosely. Some LECs dented and many vacuoles emerged, resulting in destruction of the cellular structures., Conclusion: Preservative-free 1% lidocaine may loose the junction between LECs and between the cells and the capsule, and also destroy cellular structures, resulting in degeneration and necrosis of the LECs.

The aim of this study was to expand hematopoietic progenitor cells at large scale by magnet stirred culture system. Mononuclear cell from umbilical cord blood were cultured in serum-free medium with stem cell factor, FIT-3 ligand and thrombopoietin. Firstly, the role of magnet on cell growth and colony-forming was studied by static culture on 0, 25 and 50 mT. Then the expansion multiple of cells, colony-forming and expression of surface markers were studied in magnet stirred culture by cell counting, colony-forming assay and flow cytometry. The results indicated that there was no difference in multiple of total cell expansion and numbers of hematopoietic colonies between 0, 25 and 50 mT groups and spinner groups (all p > 0.05). After 7 day cultures, the multiple of total cell expansion in magnet stirred culture was higher than that in static culture (p < 0.01). The numbers of CFU-GM (colony-forming unit-granulocyte/macrophage) and CFU-E (erythroid colony forming unit) in magnet stirred culture were higher than those in static culture, (p < 0.05). The primitive cells (CD34(+), CD34(+)/CD38(-) or CD133(+)) of the expanded cells in magnet stirred culture were less than those in static culture (p < 0.05). However, the CD184(+) or CD62L(+) expanded cells were more than that in static culture (p < 0.05). It is concluded that magnet stirred culture favors the expansion of hematopoietic progenitor cells. The results will be finally confirmed in further in vivo experiments and clinical applications.

To investigate whether lymphocyte function-associated antigen 1 (LFA-1) and very late antigen 4 (VLA-4) are involved in vasoendothelial adhesion and transendothelial migration of high proliferative potential endothelial progenitor cells (HPP-EPCs), flow cytometry was used to analyze the expression of integrin beta1 and beta2, the expression of intercellular adhesion molecule (ICAM-1, 2) and vascular cell adhesion molecule (VCAM-1) in mouse bone marrow endothelial cells (mBMECs). The adhesion and transmigration through endothelial cells of the HPP-EPCs blocked by functional grade neutralizing antibodies of VLA-4 and LFA-1 were studied in vitro. The results revealed that HPP-EPCs were positive for CD11a and CD49d in HPP-EPCs. The expression of ICAM-1and VCAM-1 of mBMECs increased after activated by IL-1beta and TNF-alpha. The results of adhesion in vitro revealed that the numbers of the adhered and migrated cells in the CD11a antibody group, in the CD49d antibody group and in the combinational antibody group were less than those in the isotype control antibody group. Furthermore, the number of adhered and migrated cells in the combinational antibody group was less than that in the CD11a or the CD49d antibody group (p < 0.05). It is concluded that both LFA-1 and VLA-4 are involved in vasoendothelial adhesion and transendothelial migration of HPP-EPCs.

The aim of this study was to establish the method of isolating and culturing endothelial progenitor cells (EPCs) from human umbilical cord blood. Mononuclear cells (MNCs) from human umbilical cord blood were cultured by using culture system supplemented with endothelial cell-conditioned medium. The obtained two types of cells were purified by picking up colonies, identified by uptake of acetylated low-density lipoprotein (Ac-LDL) and binding to lectin [Ulex European Agglatinin (UEA-1)], and were analyzed for the expression of markers by flow cytometry. The results showed that there were significant differences between two types of cells in proliferation, so they were referred as circulating angiogenic cells (CACs) and high proliferative potential endothelial progenitor cells (HPP-EPCs), respectively. They were in accordance with the standards of EPCs, could uptake DiI-Ac-LDL and bind to UEA-1, and expressed the markers of endothelial cells, such as CD31, CD144 and vWF detected by immunocytochemistry. The transcription of CD31, KDR, CD144 and ENOS in both of them could be detected by RT-PCR, but FACS analysis showed significant differences of surface marker expression between them. In conclusion, two types of EPCs are successfully obtained by culturing MNCs isolated from human umbilical cord blood using endothelial cell-conditioned medium.

Objective: To assess whether preservative-free 1% lidocaine is capable of loosing the junction between the rabbit lens epithelial cells (LECs) and between the cells and capsules and is capable of destroying the LECs, in order to provide scientific basis for pursuing safe and effective drugs to eliminate LECs in cataract surgery and to prevent capsular pacification., Methods: Lens capsule specimens were collected from 29 rabbits (58 eyes) and divided into 3 groups: balanced salt solution (BSS) group (exposed to BSS for 1 minute), lidocaine group (exposed to preservative-free 1% lidocaine for 1 minute) and the control group. Specimens were stained with trypan blue and alizarin red. Photomicrographs of each capsule were taken to observe the viability of LECs and count the number of dead LECs. The histopathologic changes of LECs treated with lidocaine were evaluated by histological method and transmission electron microscope., Results: The rate of dead LECs of the anterior and the equatorial lens capsules in control group was (1.51 +/- 0.39)%, and (1.52 +/- 0.32)%, respectively. The rate of dead LECs of the anterior and the equatorial lens capsules irrigated with BSS was (1.78 +/- 0.50)% and (1.77 +/- 0.47)%. The rate of dead LECs of the anterior and the equatorial lens capsules irrigated with preservative-free 1% lidocaine was (13.01 +/- 4.67)% and (9.59 +/- 3.35)%, respectively. The nested design ANOVA showed that the rate of dead LECs in the lidocaine group was greater than that in the control group or BSS group (P < 0.05), There was no significant difference in the number of dead cells between the anterior lens capsules and the equatorial lens capsules. After irrigated with lidocaine, cavities appeared ibetween the LECs and the capsule, cells detached from the capsule and showed vacuoles. The capsules of control group and BSS group showed a normal layer of LECs attached to the capsule. Under transmission electron microscope, in the lidocaine group, the junction between LECs and between the cell and the capsule were destroyed, many cells detached from the capsule and the rest arranged loosely. Some LECs dentes, and many vacuoles emerged, resulting in destruction of the cellular frame., Conclusion: Preservative-free 1% lidocaine may loose the junction between the LECs and between the cells and capsules, resulting in degeneration and death of the LECs.

Objective: To investigate the value of 99mTc-MIBI myocardial perfusion tomography monitoring the cardiotoxicity induced by anthracycline., Methods: Twenty-three patients with anthracycline chemotherapy were examined by electrocardiogram (ECG), myocardial enzyme (CK-MB), nuclear angiography for detecting left ventricular ejection fraction (LVEF) and 99mTc-MIBI myocardial perfusion tomography for detecting myocardial relative quantity (MRQ). These examinations were repeated after every chemotherapy., Results: The MRQ after one period of anthracycline chemotherapy was significantly lower than the pretherapy in 23 patients (P < 0.01). The MRQ significantly decreased after one period of chemotherapy in 11 patients treated by pirarubicin, in 6 by epirubicin, and 6 by mitoxantrone (P < 0.05). There was not significant change in the mean value of ECG and CK-MB after one period of chemotherapy (P > 0.05). After multiple-period anthracycline chemotherapy in 10 patients, a decrease was observed in MRQ (P < 0.01). There was not significant difference in MRQ between multiple periods and one period therapy (P > 0.05) and in LVEF in the period before and after multiple-period chemotherapy (P > 0.05)., Conclusion: 99mTc-MIBI myocardial perfusion tomography can monitor the anthracycline cardiotoxicity and its changes are earlier than LVEF's. 99mTC-MIBI myocardial perfusion tomography may be helpful to the clinical treatment for anthracycline.

Objective: To investigate the relationship between the axially lymph nodes metastases and the bone metastases in patients with the breast cancer., Methods: Lymphadenotomy of the axially lymph nodes had been performed on patients with the breast cancer, and the excised axially lymph nodes were observed by pathological examination. All patients underwent postoperatively total bone scan with single photon emission computer tomography (SPECT) periodically., Results: Among 292 patients with the breast cancer, the bone metastatic rates of low and high differentiating tumor were 27.3% (6/22) and 45.2% (122/272) respectively, the bone metastatic rates in both tumors had no difference (P > 0.1). The bone metastatic rate was 54.3% (100/184) in patients suffered from the axially lymph nodes metastases, and 25.9% (28/108) in non-metastases patients respectively, the difference of their bone metastatic rates was significant (P < 0.001). The bone metastatic rate was 48.4% (62/128) in patients suffered from the metastases of axially lymph nodes after operation less 2 years, and 67.8% (38/56) after operation more than 2 years respectively, the difference of their bone metastatic rates was not significant (P < 0.5)., Conclusions: There is no relationship between the bone matastatic rates and the pathological type of tumor. The bone metastatic rates of patients with the axially lymph nodes metastases are higher than those with non-metastases of axially lymph nodes. In addition, the bone metastatic rates of patients with the axially lymph nodes metastases increase with time prolongation.