The ligand-binding domain of VLDL receptor contains eight imperfectly similar repeats. To discuss the contribution of each repeat to ligand binding, the RT-PCR technique was used to clone the VLDLR-cDNA from the heart muscle of Chinese people. Two recombinants were further constructed, which contained the full-length cDNA of VLDLR and the mutant lacking repeats 1–5. CHO cell line was transfected with two recombinants. The expression of VLDLR gene could be detected by RT-PCR from the CHO cells transfected with pCD-VR. The results of binding experiments showed that the ability of the CHO cells transfected with the full-length cDNA of VLDL-R binding Dil-labeled β-VLDL was higher than that of the CHO cells transfected with the mutant. Our findings indicated that human VLDL-R gene could be expressed effectively on CHO cells, and the receptor was almost inactivated when repeatsl-5 were deleted.
The BALB/c mice were immunized with Hsp70 DNA and Hsp65 DNA vaccines in humanMycobacterium tuberculosis. Eight weeks after immunization, the eyeballs were removed, blood and spleen taken, and intraperitoneal macrophages were harvested. The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release to measure the phagocytic activity of the macrophages. With ELISA kit, the levels of interleukin-2 (IL-2) and interferon-γ (IFN-γ) in serum and the splenic lymphocytic cultured supernatant were detected. The results showed that after the mice were immunized with 100 μg/mouse of Hsp70 DNA vaccine intramuscularly, the splenic lymphocytic proliferating ability in the mice was significantly increased as compared with that in the control group, vector group and Hsp65 DNA vaccine group (P<0.01); The contents of NO in the intraperitoneal macrophages of the mice were significantly lower than in the control group and Hsp65 DNA vaccine group (P<0.01); The levels of serum IL-2 in the mice were significantly higher than in the control group, but there was no statistical difference between Hsp65 DNA group and vector group (P>0.05); The contents of serum IFN-γ in the mice were significantly higher than in the control group, but significantly lower than in the Hsp65 DNA vaccine group (P< 0.05). It was indicated that immunization with Hsp70 DNA vaccine could obviously enhance the immune response, but its intensity seemed inferior to Hsp65 DNA vaccine. The anti-infection mechanisms and clinical use in the future of the vaccines of Hsp70 DNA and Hsp65 DNA are worth further studying.
In order to study the effects of 1-(2, 6-dimethylphenoxy)-2-(3, 4-dimethoxyphenylethylamino) propane hydrochloride (DDPH) on proliferation and immunophenotypes of newborn rat pulmonary vascular pericytes induced by hypoxic endothelial cell conditioned medium (HECCM) from porcine pulmonary arteries, the cultured pericytes were divided into 4 groups according to the endothelial cell conditioned medium (ECCM) used: normoxic ECCM (NECCM) group, NECCM + DDPH group, HECCM group and HECCM+DDPH group. Cell culture, immunocytochemical staining, image analysis and flow cytometric method were used to investigate the effects of HECCM and DDPH on the expression of α-smooth muscle actin (α-SM-Actin) antigen, CD34 antigen, S-100 antigen and proliferating cell nuclear antigen (PCNA) and cell cycle in pericytes. The results showed that the α-SM-Actin antigen in the pericytes in HECCM group was stronger positively expressed than in the other three groups, but CD34 antigen and S-100 antigen were negatively expressed. The expression of α-SM-Actin antigen, CD34 antigen and S-100 antigen was positive in the groups of NECCM, NECCM + DDPH and HECCM + DDPH; The expression of α-SM-Actin and PCNA in HECCM group was 1. 32 times (P<0.01) and 1. 24 times (P<0.05) that in NECCM group, 1. 30 times (P <0. 01) and 1. 21 times (P<0.05) that in HECCM + DDPH group, respectively. The percentage of the cells in the GO-G1 phase in the HECCM group was lower by 11. 7 % and 9. 1 %, in S phase higher by 5. 6 % and 4. 2 %, in G2-M phase higher by 6. 1 % and 4. 9 % than in the groups of NECCM, HECCM+DDPH, respectively. The inhibitory rate of DDPH on the increased α-SM-Actin and PCNA syntheses in pericytes induced by HECCM were 23. 4 % and 17. 1 % respectively. The inhibitory rate on the increased pericytes from GO-G1 phase to S phase was 8. 3 %. These results suggest that DDPH can directly inhibit pericytes from proliferation and immunophenotypical transformation of smooth muscle-like cells induced by HECCM.
In order to study the fate of human follicle-stimulating hormone (FSH) when hormone binds to its receptor, a quick biochemical method that can differentiate between the surface-bound and internalized hormone was used to determine the internalization induced by FSH in cultured both porcine granulosa cells and Chinese hamster ovary (CHO) cells expressing recombinant porcine FSH receptor. The results showed that FSH was slowly internalized, and the internalized radioactivity (acid resistant) reached a peak 10–12 h after addition of125I-hFSH. It was suggested that FSHR do not get internalized rapidly under physiological circumstances precisely because the appropriate sequences are absent.
A novel variant of human vascular endothelial growth factor (h’VEGF165) cDNA was amplified by nested PCR method from the HL601 cells and was cloned into a eukaryotic expressing vector pcDNA3 to construct a recombinant plasmid pCD-h’VEGF165. The amplified h’VEGF165 cDNA fragment was identified by enzyme digestion and DNA sequencing methods. Also, wild-type hVEGF165 cDNA was obtained, identified and cloned into a eukaryotic expressing vector pcDNA3 by using the same methods. The results of DNA sequencing showed that h’VEGF165 cDNA cloned from HL601 was 600 bp in size with 8 % of the base sequence in h’VEGF165 cDNA being changed as compared with the base sequence in the wild-type hVEGF165 cDNA. The results of sequencing of hVEGF165 which was cloned from HL60 by us were consistent with the reports completely.
The mechanism of vascular endothelial growth factor (VEGF) on the prevention of restenosis after angioplasty was investigated. The cultured vascular endothelial cells (VEC) were incubated with the conditioned medium (CM) from vascular smooth muscle cells (VSMC) infected with recombinant adenoviruses containing the hVEGF165 gene. To observe the effects of VEGF on proliferation and NO, ET, 6-keto-PGF1α secretion of VEC, WST-1 method, Griess method and radioimmunoassay were used respectively. The PDGF-B mRNA transcription in VECs was detected by RT-PCR. It was showed that NO, 6-keto-PGFla and OD value were markedly increased in a dosedependent manner in the VEGF-treated groups as compared with those in the control group, while ET and PDGF-B mRNA were significantly decreased in the VEGF-treated groups (P<0.05 orP< 0. 01). Adenovirus vector mediated hVEGF165 gene could promote the proliferation of VECs and improve NO, PGI2 secretion, inhibit ET secretion and PDGF-B mRNA transcription in the VECs. The above results offered further theoretical evidence for VEGF on the prevention of restenosis after angioplasty.
In order to investigate the immunological damage in rat immunized with ATI-receptor peptide. 18 male Wistar rats were divided into two groups: immunized-group (n=12), each rat was immunized with 150 μg AT1-receptor petide coupled to bovine serum albumin, together with Freund’s adjuvant. Control group (n=6), sham-immunized, “immunized liquid” was same as immunized-group except ATI-receptor peptide. Systolic blood pressure (SBP) was measured by using the tail-cuff technique, antibody against ATI-receptor peptide detected by using ELISA method, and left ventricular myocardium and renal cortex sections were observed under light and electron microscopy. There was no significant difference in SBP and light microscopic observation of the tissue sections between the immunized-group and control group. The O.D. value of anti-ATI-receptor peptide antiserum was significantly higher in the immunized-group than in the rats before immunization and control group (P<0.01). Positive rate in the immunized-group was 100%, while 0% in the control group. Ultramicroscopic morphology showed potential myocardial injury, including: increase in number of mitochondria, swelling of many mitochondria with reduction in number or absence of their cristae and cristolysis, disorder of the cardiac myofibrils, and myofibrillar disruption and myocytolysis. And lysosomes were increased in renal tubular epithelia. The ATI-receptor peptide could induce to generate the antibody against ATI-receptor peptide and lead to myocardial and renal damage in rats.
The apoptosis and the expression of p53, bcl-2 and Bax in myocytes of chronic rapid ventricular pacing-induced congestive heart failure (CHF) in rabbits were investigated. The CHF rabbit model (P,n=7) was established by chronic rapid ventricular pacing for 3 weeks. By using TUNEL technique the apoptosis in the myocytes in the rabbit model was studied and the expression of p53, bcl-2 and Bax in myocytes was detected by using immunohistochemical method. Sham-operated (C,n = 9) group served as control group. The results showed that there were about 4033±884. 56 apoptotic cells/106 myocytes in P group, but no apoptotic cells were found in C group. Myocytes positive for p53 immunoreactivity (18.86±8.48 vs 5.06±0.87,P<0.01) and positive for Bax immunoreactivity (7. 15±1. 91 vs 0. 43±0. 09,P<0.01) were increased in P group as compared with those in C group, while the myocytes positive for bcl-2 immunoreactivity (7. 08±1.05 vs 14. 97±4.47,P< 0.01) and the ratio of bcl-2/Bax were decreased in P group as compared with those in C group. Apoptosis was involved in the development of CHF induced by continuously rapid ventricular pacing in rabbit. The expression of p53 and Bax was increased, while the expression of bcl-2 was inhibited. These might play an important role in the acceleration of the apoptosis.
In order to study the effect ofErigeron Breviscapus (EB) on proliferation of pulmonary artery smooth muscle cells (PASMC) in hypoxic porcines, immunohistochemical and MTT methods were employed to measure the proliferation of PASMC. It was found that the proliferation of PASMC in porcines was obvious, and the expression of proliferating cell nuclear antigen (PCNA) was significantly high within 48 h after exposure to hypoxia. The EB could inhibit the proliferation and the expression of PCNA in PASMC under hypoxia, but it had no effect on the proliferation and expression of PCNA in PASMC under normal condition. The EB could inhibit the proliferation and the expression of PCNA in PASMC induced by phorbol 12-myristate 13-acetate (PMA), an agonist of PKC in normal and hypoxic conditions. It was concluded that the hypoxia could enhance the proliferation and expression of PCNA in PASMC. The EB can inhibit the proliferation and expression of PCNA in PASMC under hypoxia through PKC-signal way. The EB may be used in treating the pulmonary hypertension by inhibiting the proliferation of PASMC and the pulmonary vascular remodeling.
In order to study the significance of flow cytometry immunophenotyping in the diagnosis of acute leukemia, CD45/SSC gating multiparameter flow cytometry (FCM) was utilized to analyze the immunophenotypes of 139 cases of acute leukemia. 139 cases of acute leukemia were enrolled in our hospital from April 1998 to April 2000. Morphological analysis and FCM immunophenotypic tests were conducted on all cases. Our results showed that CD45/SSC gating multiparameter flow cytometry immunophenotyping could reflect the origin of leukemic cells specifically. It is one of the important methods for the diagnosis of ALL, AML, and HAL. CD45/SSC gating multiparameter FCM analysis is a good technique for immunophenotyping. FCM immunophenotypic analysis can help improve the diagnosis and classification of acute leukemia, and extend the use of FCM in clinical practice.
The mechanism of chemotherapeutic drug-induced apoptosis in leukaemic cells was studied to further investigate whether Fas/FasL system was involved in apoptosis induced by chemotherapeutic drugs and assess their effects when used in combination with soluble FasL (sFasL). The expression of Fas on human leukaemic cell lines K562, HL-60 and U937 treated with daunorubicin (DNR) or cytosine arabinoside (Ara-C) was detected by using flow cytometry. The activities of sFasL, DNR and Ara-C inducing apoptosis of leukaemic cells, in the absence or presence of neutralizing anti-Fas IgG antibody, were detected by using flow cytometry and TUNEL. The results showed that flow cytometric profiles of K562, HL-60 and U937 cells treated with DNR or Ara-C failed to show any significant increase in Fas expression over 18 h (P>0.05). Anti-Fas monoclonal antibody (IgG) could not block the apoptosis in leukaemic cells induced by DNR or Ara-C, but could block the apoptosis induced by sFasL. A role of sFasL in a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs was revealed. It was concluded that chemotherapeutic drug-induced apoptosis in human leukaemic cells (UG37, HL-60) is independent of the Fas/FasL system, but combination of sFasL and drug treatment produces a synergistic cytotoxic effect on human luekaemic cells.
The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on N1H3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas+ Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1 cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8. 5 × 105 colony-forming-unit (CFU)/ml. The NIH3T3 cells transfected by above supernatants had a higher level of mFasL (53. 81 ±6.9 %), and significantly induced the apoptosis of Fas+ Yac-1 cells (56.78±4. 5 %), as both were cocultured for 5 h at 1:1 ratio, whereas it is 7. 08±3. 4 % in control group (P<0.01). Supernatant from PLFIN-PA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.
To study the preparation and cleavage of hairpin ribozyme (HpRz) directed against the transcript of HBV core regionin vitro, HRz gene designed by computer targeting the transcript of HBV core gene was cloned into the vector p1. 5 between 5′-cis-Rz and 3′-cis-Rz.32P-labeled HpRz transcript proved whether the vector fit for the preparation of hairpin ribozyme.32P-labeled pKC transcript containing HBV core region as targets-RNA was transcribed by using T7 RNA polymerase and purified by PAGE. Cold HpRz transcript was incubated with32P-labeled target-RNAs under different conditions and radioautographed after denaturing polyacrylamide gel electrophoresis. The results showed that HpRz had the ability of cleavage at 37°C and 12 mmol/L MgCl2 and the design of ribozyme was correct. It is concluded that HpRz preparedin vitro possesses specific catalytic activity, indicating that it is possible for HpRz to intracellularly inhibit the replication of HBV. It may be developed into a nucleic acid drug in the treatment of hepatitis B in the future.
The role of interleukin-6 (IL-6) in the growth of an androgen-independent prostate cancer cell line (PC-3m) was defined and the effect of dexamethasone, which was previously shown to modulate IL-6/IL-6 receptor (IL-6R) on this procedure was investigated. By using a pretty sensitive and specific enzyme immunoassay (ELISA), it was found that PC-3m produced certain IL-6, but there was no difference in IL-6 secretion between the group with or without dexamethasone treatment. It was also found that PC-3m cells could not be stimulated to grow by exogenous IL-6 (P>0. 05), while it could be inhibited to grow by anti-IL-6 monoclonal antibody and dexamethasone with a dose-dependent fashion. Our observation indicated that IL-6 acted as an autocrine growth factor for PC-3m, and dexamethasone could inhibit cell proliferation by a mechanism independent of its effect on IL-6 mRNA expression.
To explore the relationship of angiogenesis-related angiopoietin-2 gene and its receptor Tie2 with angiogenesis and the biology, of hepatocellular carcinoma (HOC), angiopoietin-2 gene, Tie2 and CD34 protein expression in 22 resected HOC, 8 cirrhotic and 8 control liver specimens were investigated byin situ hybridization and immunohistochemistry respectively, and the level of angiopoietin-2 and Tie2 expression in HCC were compared in terms of tumor biological parameters. It was found that CD34 was not expressed in control liver, expressed scarcely in cirrhotic liver (17. 8±13.5/HP), but intensively expressed in HCC (86. 3±34. 8/HP,P<0.01). Tie2 receptor was not expressed in controls, expressed at low level in cirrhotic liver (11. 3±8.7/HP), while strongly positive in the microvascular endothelia of HCC (52. 4±16. 7/HP,P<0.01). The level of Tie2 receptor expression in HCC was closely related with tumor diameter, angiogenesis and portal invasion. Angiopoietin-2 gene was not expressed in control liver, expressed mildly in cirrhotic liver (11.2±9. 7/HP), but extensively in tumor zone (36. 4±17. 5/HP), the level of angiopoietin-2 expression was closely related with angiogenesis, portal invasion and histological grading of HCC. It is concluded that angiogenesis is increased in HCC; angiopoietin-2/Tie2 expression in human hepatic carcinoma is closely related with angiogenesis, which are probably involved in the HCC angiogenesis regulation, promoting the development and metastasis of human hepatic cancer.
The role of CyclinD1 and estrogen receptor (ER) in the process of proliferation and metastasis of breast neoplasm and their relationship were studied. The expression levels of CyclinD1 and ER in the tissue samples were detected by using flow cytometry and L SAB immunohistochemistry staining, respectively. The results showed that CyclinD1 and ER expression levels in breast cancer were significantly higher than in benign breast neoplasm (P<0.05). The CyclinDl expression levels in stage I was much lower than in stages II, III, IV (P<0.05). The positive rate of ER was not related with tumor size, lymph node metastasis and TNM stage (P>0.05), but the CyclinD1 expression level in ER (+) group was significantly higher than in ER (-) group (P<0. 05). It was concluded that CyclinDl expression level might be obviously related with the proliferation and metastasis of breast neoplasm and ER.
The targeting of antineoplastic agents to restricted anatomic sites and specific target cells have been challenged clinicians all the time in cancer chemotherapy, which resulted in recent efforts to focus the effects of existing antitumor agents and treatments on tumor cells and spare their effects on normal cells. The drug-carrier complex, adriamycin carried by magnetic albumin microspheres (ADM-MAM) was prepared by using our discovered new and modified method. The physical feature of the prepared drug-carrier microspheres was much better than by the traditional method in comparison. The successful preparation of the drug-carrier complex, ADM-MAM, is one of the key steps for our later further researches in the targeted chemotherapy.
In order to evaluate the value of the ultrasonography in the diagnosis of tumor of the knee and its clinical implication, 67 patients with clinically suspected bone tumor of the knee were examined by ultrasound. The ultrasonographic characteristics of different bone tumors were studied and compared with the results of pathologic characters after operation. Ultrasonography can readily visualize the bony destruction and the pathologic change of the periosteum and the soft tissue related to bone tumor. Fifty-two cases of malignant bone tumors and 15 cases of giant cell tumors were diagnosed by ultrasonography. Pathologically, there were 54 cases of malignant bone tumor and 13 cases of giant cell tumor. It was concluded that ultrasonographic examination might be a useful method for the diagnoses of bone tumor of the knee and play an important role in guiding needle biopsy and electing operative method and approach.
The incidence of the co-infection of hepatitis G virus (HGV) and hepatitis C virus (HCV) and its clinical implication was investigated and the difference in the positive rate of HGV RNA and HCV RNA between plasma and peripheral blood mononuclear cells (PBMCs) observed. By using reverse transcriptase polymerase chain reaction (RT-PCR) assay, HCV-RNA and HGV-RNA in plasma and PBMCs of 72 patients with hepatitis C was detected. It was showed that HGV RNA was positive in plasma of 11 patients, in PBMCs of 15 patients, and simultaneously in both of plasma and PBMCs of 10 patients with the co-infection rate being 22.2 %. Nine patients were both HGV RNA and HCV RNA positive in plasma, 11 patients were both HGV RNA and HCV RNA positive in PBMC, and 6 patients were both HGV RNA and HCV RNA positive in both plasma and PBMC with the positive rate being 12.4 %, 15.3% and 8.3 % respectively. The positive rate of both HGV RNA and HCV RNA in PBMCs was higher than in plasma. It was concluded that the HGV co-infection rate in the patients with hepatitis C was 22.2 %. Simultaneous examination of plasma and PBMC can improve clinically detectable rate.
To determine the CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome (HFRS) and its clinical implications, double immunofluorescence technique and flow cytometry were used. There was no significant difference among the severe group, mild-moderate group and normal control group in the CD+CD30 T lymphocyte subset. While the CD4+CD30− T cells of HFRS patients were increased and the difference between severe group and mild-moderate group or normal control group were very significant (P<0.01) and the difference between the mild-moderate group and normal control group was also significant (P<0.05). The CD8+CD30− T cells were increased while the CD8+CD30- T cells decreased obviously in HFRS patients, and the differences among three groups in both subsets were very significant (P<0.01). The results showed that the humoral immunity and cellular immunity are overactive in HFRS patients during acute phase. The loss of balance between T lymphocyte subsets may play an important role in the pathophysiology of HFRS and is closely correlated with the severity of the HFRS.
To investigate the clinical significance of magnetic resonance imaging (MRI) of bone marrow in patients with acute leukemia, the femoral and pelvic marrow were evaluated by using MRI with a T1-weighted spin-echo (SE) method and a short T1 inversion recovery (STIR) technique. Normal bone marrow examination was performed with coronalT1-weighted MRI of pelvis and femurs, and showed persistent red marrow. There was a bright signal of fatty marrow in the femoral epiphyses and apophyses. MRI pattern of bone marrow in the 54 cases of acute leukemia showed abnormal signal patterns of femoral and pelvic marrow: (1) grade I (n=4),(2) grade II (n = 11), (3) grade III (n = 8), (4) grade IV (n=17), and (5) graded V (n=14). Leukemic cells had infiltration onseted by red marrow in adult patients with leukemia. The marrow of femur had infiltration from diaphysis to epiphysis, and to femoral head and greater trochanter. The lower grades (grade IV, V) of leukemic marrow supported the diagnosis of AML in MRI, which achieved higher complete remission. The adult patients with ALL had higher grades (grade I–III) in MRI. Our findings indicated that MRI of femoral marrow is an important tool for accurate diagnosis and management of patients with leukemia that may function as an adjunct to bone marrow aspiration and biopsy. The pattern of MRI in patients with newly diagnosed leukemia predicted the prognosis and CR of leukemia.
The backscatter from sonicated albumin microbubbles (Albunex) was analyzed using acoustic densitometry in anin vitro pulsatile heart model to evaluate the effects of pressure on the backscatter from Albunex, and the cardiac cyclic changes of intracardiac backscatter from sonicated albumin microbubbles in 16 healthy persons were analyzed. It was found that the Albunex microbubbles were compressed in systole and decompressed in diastole, causing corresponding changes of backscatter in cardiac cycle. Although the intensities of backscatter in diastole and systole were related to the concentration of microbubbles, the concentration of microbubbles had no effect on the difference of end-diastolic and end-systolic backscatter. The difference of the backscatter was highly correlated with end-systolic pressure (r=0.96,P = 0.001). In human studies, we also observed same intracardiac cyclic changes of backscatter from sonicated albumin microbubbles. Our study indicates that it is possible to evaluate the intracardiac pressure non-invasively by analyzing the intracardiac backscatter from the microbubbles with acoustic densitometry.
In order to evaluate the predictive value of maternal plasma fibronectin (FN) concentration at 24–34 weeks on fetal intrauterine growth retardation (IUGR), a prospective double-blinded study was performed. The maternal plasma FN concentrations were measured by using a rate nephelometric procedure in the 130 initial normal nulliparous pregnant woman at 24–34 gestational weeks. The outcome of pregnancies and birth weight of their infants were followed up. IUGR was defined as that the birth weight was less than the 10th percentile for gestational age. The receiver operating characteristic curves and predictive values of FN predicting on outcome of pregnancy with IUGR were analyzed. The results showed that: (1) In a cohort of 130 initially normal nulliparous pregnant women, IUGR occurred in 14 cases during the follow-up; (2) The plasma FN levels in the women with I-UGR (467. 58±l04. 43 mg/L) were significantly higher than in the normal control group (299. 44±105. 55 mg/L,P<0.01). However, there was no significant difference in the mean maternal age, gravidity, sampling gestational ages, delivering gestational ages between the two groups (P>0.05); (3) The areas under ROC curve for predicting the outcome of pregnancy in IUGR was 0.893; (4) At the cut point of 475 mg/L FN level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index for predicting the outcomes of pregnancy in IUGR were 57.14 %, 95.69 %, 61.54 %, 94.87 %, 0.5455 respectively. It was concluded that the maternal plasma FN might be used as an earlier predictor for screening of IUGR.
The isolated cardiac myocytes of rats were immediately infected by cosackievirus B3 (CVB3) to investigate the effects of such procedure on the cell cycle, apoptosis and intrazellular ionized calcium (Ca2+ i) of cardiac myocytes. Newborn Balb/c murine cardiac myocytes were cultivated, then infected by CVB3. Intracellular Ca2+ i was measured by flow cytometer. The calcium in the medium for culturing cardiac myocytes was detected by using atom absorb spectrum test. It was found that CVB3 could markedly inhibit the differentiation and proliferation of the infected cardiac myocytes and induce the apoptosis. The intracellular Ca2+ i level in the infected group was significantly higher than in the control group (P<0.01). The calcium concentration in the medium for culturing cardiac myocytes in the infected group was significantly lower than in the control group (P<0.05). It was suggested that the apoptosis and intracellular calcium overload of the CVR3-affected cardiac myocytes are likely to play an important role in the pathogenesis of viral myocarditis.
In order to improve the curative effect of onychomycosis, the factors influencing the therapeutic effects were investigated. 545 cases including 245 males and 300 females, who were diagnosed both clinically and mycologically, were treated by Intraconazole with intermittent pulse therapy. The therapeutic effects were judged by the following observations regularly and analyzed from the factors as follows: age; growing speed of nails; accompanied diseases; family history; trauma of nails; infection ways of the pathogens; manifestation of the injury; pathogens; duration of the treatment. The results showed that the recovery rate was higher in younger patients (P<0.01) with a quicker recovery rate (P<0.001), and a lower recurrent rate (P<0.01), as well as in those with quicker growing speed of new-born nail. Also the patients with WSO and DLSO manifestation had a higher recovery rate. The patients with onychomycosis caused by T. rubrum had a higher recovery rate (P< 0.01 to 0.001) no matter whether to prolong the treatment duration. The patients with diabetes mellitus or hyperhidrosis, as well as with positive family history or basic nail diseases such as trauma and paronychia, had a lower recovery rate and the curative effects were not satisfactory. It was concluded that although the single and some DLSO-manifestation nail injury could be cured by internal and external treatments with the help of removing the sick nail and the duration of the treatment could be shortened. The treatment duration should be prolonged in order to increase the curative effects and decrease the recurrence under such conditions as following: old patients above 60 years; patients with low-growing-speed new-borne nails; patients with thumb and big toel injury and ingrowing nail; patients with diabetes mellitus, hyperhidrosis or Renauld’s phenomenon; patients with nail trauma before or during the treatment; patients with PSO or TDO manifestation; patients with onychomycosis caused by Candida or Aspergillus; patients with abnormal new-borne nails of abnormal color, coarse surface or abnormal thickness.
The apoptosis of lens epithelial cells (LECs) induced by ultraviolet and the expression of P53 were investigated. Wistar rats received 100 mW/m2 ultraviolet irradiation (UVR) (λ=280 nm-315 nm) for 15 min. One, 6, 24 h after irradiation the lens capsules were dissected. The percentages of apoptotic cells were evaluated by the TdT-dUTP terminal nick-end labeling (TUNED technique and the expression of P53 was detected by using immunohistochemical assay. The results showed that the percentages of TUNEL-positive nuclei at 24 h after irradiation was significantly higher than in the control group and those 1 h. 6 h after irradiation. The percentages of P53-positive cells at 6 h, 24 h after irradiation were significantly higher than in the control group and those 1 h after irradiation. It was concluded that UVR could induce the apoptosis of lens epithelial cell. The expression of P53 might be responsible for the apoptosis of lens epithelial cells.