In vitro suppressive effect of angelica polysaccharide on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells / 中国实验血液学杂志

The objective of study was to investigate the in vitro suppressive effect of angelica polysaccharide (APS) on human cytomegalovirus-induced apoptosis via direct infection in CHRF-288-11 cells. HCMV AD169 directly infected CHRF-288-11 were cultured in vitro, APS in different doses were added on day 3 after the infection of virus. Cells of every group were collected at different time points. HCMV DNA of cells were detected by using polymerase chain reaction and the apoptotic cells were examined by using Hoechst staining, MTT assay, DNA fragmentation assay and flow cytometry. The results showed that the APS to some extent inhibited the apoptosis of CHRF cells infected by HCMV in vitro in a dose-dependent manner. The expression of HCMV IEA in CHRF-288-11 cells was found by PCR amplification. Morphology observation, flow cytometry assay and DNA fragmentation assay revealed the existence of apoptosis. With the dose decrease of APS added to the infected CHRF cells, the percentage of apoptotic cells increased. It is concluded that the HCMV AD169 can infect CHRF cells directly in vitro and decrease cell viability. HCMV AD169 infection increases the apoptosis of CHRF cells in time-dependent manner. When APS was added to the CHRF cells infected by HCMV AD169 in vitro, the viability of CHRF cells increase, which indicated that APS to some extent protects the CHRF cells infected by HCMV. APS suppresses the cytomegalovirus-induced apoptosis in CHRF cells directly infected in vitro in dose-dependent manner.

Human cytomegalovirus induces apoptosis of human promyelocytic leukemic cells via direct infection in vitro / 中国实验血液学杂志

The study was purposed to investigate the proliferation and the suppression effect of human cytomegalovirus (hCMV) on human promyelocyte cell line HL-60, and to study whether hCMV can induce apoptosis of HL-60 via direct infection in vitro and its mechanism. Promyelocyte cell line HL-60 and hCMV AD169 strain were co-cultured. PCR was used to detect the direct infection of HL-60 cells by hCMV IEA expression. The apoptosis cells were analyzed by morphologic observation, DNA ladder formation, flow cytometry with Annexin V/PI stain. The results indicated that hCMV AD169 suppressed the differentiation and proliferation of HL-60 cells in vitro significantly (P < 0.05). The suppression was dose-dependent. hCMV DNA was successfully detected in HL-60 cells of viral infection groups by PCR. The apoptotic cells were confirmed by morphologic observation and DNA ladder formation. The results of flow cytometry showed that the percentage of apoptotic cells increased along with the increase of hCMV titer and the time after infection. It is concluded that the promyelocyte can be infected directly by hCMV AD169 strain. hCMV AD169 strain inhibited the differentiation and proliferation of promyelocyte. The apoptosis of HL-60 cells can be induced by hCMV infection. With the increase of viral infectious titer and the time after infection, the percentage of apoptotic cells also increase and produce in dose-dependent and time- dependent manner. Induced apoptosis may be one of the mechanisms of granulocytopenia induced by hCMV infection.

Correlation between plasma leptin level and premature infant weight loss / 中华儿科杂志

<p><b>OBJECTIVE</b>Leptin is an adipocyte-derived hormone regulating body weight and energy balance in animals and human being. Although the physiological functions of leptin in human are still unclear, its secretion is closely related to fat mass and energy expenditure in both adults and children. This study investigated whether the plasma leptin level was reduced in connection with the weight loss during the neonatal period and try to find out the role of leptin in body weight regulation and energy balance of premature infants.</p><p><b>METHODS</b>The radioimmunoassay was used to determine the plasma leptin concentration. The first blood samples were obtained at the delivered, and then collected the samples every two days until the infants' body weight recovered to the birth weight or above. At the same time, the essential fluid and energy for the patients were supplied to keep their physiological functions. One person was appointed to take responsibility to examine the body weight, body length and head circumference. Then computed out their Kaup index from the first day to the seventh or twelfth day.</p><p><b>RESULTS</b>A total of 26 premature infants were selected into the study, of which 14 cases were male and 12 female, and their gestational age ranged from 30 to 36 weeks. There was a significantly positive correlation between the premature newborns' body weight loss and their plasma leptin levels (the 1st day: n = 26, r = 0.766; the 3rd day: n = 26, r = 0.636; the 5th day: n = 26, r = 0.629; the 7th day: n = 26, r = 0.717; the 9th-12th day: n = 24, r = 0.587; P < 0.01). The time of body weight loss and the plasma leptin level which declined to extremely low were positively correlated. (r = 0.611, P < 0.01). The time when body weight loss declined to extremely low in 26 premature infants ranged form the 3rd to the 9th day after birth [(5.2 +/- 1.6) day], and that of the plasma leptin levels ranged form the 3rd to the 8th day after birth (4.7 +/- 1.4) day. The maximal ranges of the body weight loss and the plasma leptin decrease in 26 premature infants were (6.5 +/- 3.0)% and (59.6 +/- 11.3)%, respectively. In addition, there were significantly positive correlations among the plasma leptin level, the premature newborns' body length (the 1st day: n = 26, r = 0.609, P < 0.01; the 3rd day: n = 26, r = 0.419, P < 0.05; the 5th day: n = 26, r = 0.583, P < 0.01; the 7th day: n = 26, r = 0.626, P < 0.01; the 9th-12th day: n = 24, r = 0.482; P < 0.05), and the Kaup index (the 1st day: n = 26, r = 0.634; the 3rd day: n = 26, r = 0.534; the 5th day: n = 26, r = 0.542; the 7th day: n = 26, r = 0.611; the 9th-12th day: n = 24, r = 0.539; P < 0.01). Although the head circumference correlated positively with the plasma leptin level at the first week after the delivery (the 1st day: n = 26, r = 0.580, P < 0.01; the 3rd day: n = 26, r = 0.417, P < 0.05; the 5th day: n = 26, r = 0.426; P < 0.01). There was a lower correlation between them one week after the delivery (the 7th day: n = 26, r = 0.369; the 9th-12th day: n = 24, r = 0.323; P > 0.05).</p><p><b>CONCLUSION</b>There was a significantly positive correlation between the plasma leptin level and the premature newborns weight loss. Leptin may participate in the regulation of energy balance and body weight of premature infants during neonatal life. Leptin may play an important role in growth and development of premature infants.</p>

Human cytomegalovirus aggravates apoptosis of human megakaryocytes via direct infection in vitro / 中国实验血液学杂志

The megakaryocyte and platelet lineage may be one of the major sites of human cytomegalovirus (HCMV) infection. However, whether HCMV aggravates apoptosis in normal megakaryocytes was not well investigated. Megakaryocytic cell line CHRF-288-11 and HCMV AD 169 strain were co-cultured in this study. PCR was used to detect the direct infection of the cells by HCMV IEA expression. The apoptotic cells were analyzed by morphologic observation, DNA ladder formation, annexin V/PI and PI assay with flow cytometry. The results showed that HCMV significantly inhibited the growth of CHRF cells in three different concentrations of viral infection groups (10(-3), 10(-2), 10(-1)). The viability levels in each infection groups were 77%, 73% and 68% respectively after incubation for 7 days, compared with 98% in the control group. Using annexin V/PI with flow cytometry, it was shown that the percentages of apoptotic cells viral infection in groups (10(-3), 10(-2), 10(-1)) were (21.3 +/- 2.49)%, (25.8 +/- 3.65)% and (31.4 +/- 3.91)% at 7 days after infection, while the control was (3.68 +/- 1.47)%. The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, PCR detection also showed the direct infection by identification of HCMV IEA expression in CHRF cells. This study suggested that HCMV could directly infect megakaryocytes and aggravated apoptosis in HCMV-infected megakaryocytes.

Clinical investigation on the treatment of HCMV hepatitis in children / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effective therapeutic method of human cytomegalovirus (HCMV) hepatitis in children.</p><p><b>METHODS</b>Twenty-five children with HCMV hepatitis were randomly assigned to a treated group (n=13) or a control group (n=12). Both groups were treated with prednisone, glucurone, luminal and Xiaoyanlidanpian. But the treated group was given ganciclovir (GCV) + intravenous immunoglobulin (IVIG) in addition. Each infant of the two groups was checked for blood routine, liver function and HCMV copy numbers on admission and before discharge. They were seen at the third, sixth and ninth month after discharge. On each visit blood specimens were collected for HCMV copy numbers (fluorescence quantitative PCR, FQ-PCR).</p><p><b>RESULTS</b>The viral load of the treated group decreased significantly. A significant difference in viral copy numbers was found between the two groups on admission, discharge, and third, sixth and ninth month after discharge (P less than 0.001). The number of HCMV DNA copy fell to 10(3) copies/ml on discharge while that of the control group fell to the same level after the third month. The differences between the two groups in the length of hospitalization, time of initial jaundice disappearance and complete disappearance were statistically significant (P less than 0.05). The need for transfusion in the treated group was significantly less than that in the control group (chi-square=4.012, P less than 0.05).</p><p><b>CONCLUSION</b>Combination of GCV with a high dosage of IVIG to treat HCMV active infection could decrease viral load remarkably; The duration of disease, severity of symptoms, degree of anemia and the need for blood transfusion were reduced. No adverse effects related to the combination of GCV with IVIG therapy were observed.</p>

Effect of human cytomegalovirus on hematopoietic system / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system.</p><p><b>METHODS</b>Semi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique.</p><p><b>RESULTS</b>HCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P < 0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10(3) plague forming unit (PFU), 10(4) PFU and 10(5) PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10(4) PFU group was also identified by immunohistochemical staining.</p><p><b>CONCLUSION</b>HCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia.</p>