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BACKGROUND: The proliferation of peripheral blood stem cells among peripheral blood mononuclear cells (PBMCs) invitro remains unclear. There is no optimal marker for tracing PBMCs transplanted in vivo.OBJECTIVE: To observe the degree of PBMC proliferation in stem cell medium by EdU labeling and to explore thefeasibility of EdU-labeled peripheral blood stem cells.METHODS: New Zealand rabbit PBMCs were isolated and cultured for 1 to 5 days in stem cell medium supplementedwith EdU. The cells were observed and counted at 0, 1, 2, 3, 4 and 5 days in culture. The cells were harvested at eachtime point and stained with EdU fluorescent reagents. Then, confocal microscopy and flow cytometry were used to detectEdU-labeled cells.RESULTS AND CONCLUSION: (1) Freshly isolated rabbit PBMCs were rounded and showed clear outline. After 1 dayculture, most of the cells were suspended in the medium, spherical or round. There were also a few cell clusters andadherent cells scattered in a triangle or polygon shape; after 2 days culture, more cell debris were observed, and mostcells were round; when cultured for 3-5 days, increased cell debris, smaller cell mass and decreased cell densitysignificantly were observed. (2) With the prolongation of culture time, the cell count decreased gradually. (3) Whencultured for 1 day, EdU labeled cells in red were scattered. The number of cells marked with EdU red label increasedsignificantly at day 2 and remained unchanged after 3 days of culture. At 5 days of culture, the number of red cellsmarkedly decreased; the highest positive rate of EdU-labeled cells was (2.38±0.10)% at 2 days after culture. To conclude,these results showed that the proportion of proliferating cells in rabbit PBMCs was very low. EdU is capable of labelingproliferative cells among PBMCs.
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Objective To observe the clinical effects of different fluid therapies in treating severe preeclampsia complicated by ascites. Methods Between Jan. 2010 and Dec. 2012, patients with severe preeclampsia complicated by ascites in Shenzhen Maternity and Child Healthcare Hospital were included in this study. The treatment group ( n=55 ) were given intravenous drip of 6% hydroxyl starch 130/0. 4 plus furosemide, and the control group (n=52) received intravenous drip of 5%human serum albumin plus furosemide. The mean arterial pressure, respiratory rate, heart rate, oxyhemoglobin saturation, colloid osmotic pressure, hematocrit and the incidence of acute pulmonary edema were observed and compared between the two groups. Results Twenty-four hours after cesarean section, the mean arterial pressure of the treatment group was significantly lower than that of the control group, whereas heart rate and oxyhemoglobin saturation were significantly higher (all P0. 05). The average duration of edema after treatment was significantly shorter in the treatment group [(2. 43±0. 37) d versus (3. 74±0. 59) d, P<0. 01]. Conclusion 6% hydroxyl starch 130/4. 0 plus furosemide can effectively elevate the colloid osmotic pressure, resolve edema, improve hypovolemia, sustain oxygen supply to the organs and decrease the complication of pulmonary edema in patients with severe preeclampsia complicated by ascites.
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Objective To investigate the clinical value of serum procalcitonin (PCT) in the diagnosis and treatment of the patients with high fever of acute illness.Methods A total of 285 febrile patients suffered from acute illness were divided into the conventional group (n =122) and the PCT group (n =163).The routine blood examination and serum level of C-reactive protein (CRP) detection were carried out for the dignosis of these febrile patiens of two groups,and in addition,serum PCT determination was carried out in the patients of PCT group.The diagnosis was comnfirmed by etiology or immunologic assays.Routine guidelines of administrating antibiotics were used for treatment in the conventional group.Whereas the antibiotic therapy given to patients of PCT group was guided by the serum PCT levels.Two groups were compared in respect of the antibiotics costs,duration of treatment with antibiotics,percentage of patients treated with antibiotics,efficiency of antibiotics treatment and the mortality rate.The patients of PCT group were further divided as per inflammatory markers of bacterial infection into three subgroups,namely bacterial infection group,viral infection group and non-infection group.At last,the relationship between serum PCT level and the prognosis of patients was analyzed.Results The cost of antibiotics,percentage of patients treated with antibiotics,and course of antibiotics treatment were (Y) (3586.5 ± 703.3),95.08％ and (15.01 ± 11.21) days,respectively in conventional group,whereas (Y) (1871.2 ± 433.5),54.60％,(11.22 ±7.10) days in PCT group with statistically significant difference between two groups (P ＜ 0.01),but there were no significant differences in clinical efficiency and mortality between groups (P ＞ 0.05).Serum PCT (1.12 ± 0.88) ng/ml in bacterial infection group was higher than that in both virus infected group (0.21 ± 0.1 1) ng/ml and non-infected group (0.18 ± 0.13) ng/ml.There was no statistics difference in serum PCT level between virus infected group and non-infected group (P ＞ 0.05).The diagnosis of bacterial infection with serum PCT was better than other inflammatory markers because serum PCT had high sensitivity,specificity,positive predictive value (PPV) and negative predictive value (NPV).Conclusions Serum PCT level was reliable to differentiate the nature of acute infection with high fever and to evaluate the prognosis by emergency physicians.There was an important significance for rational use of antibiotics by the guidance of PCT levels.
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Objective To discuss the clinical features,management,pregnancy outcome and prognosis of obstetric mirror syndrome.Methods The clinical data of 12 cases with obstetric mirror syndrome at Shenzhen Maternity and Child Healthcare Hospital from April 2008 to December 2010 were collected to retrospectively analyze the clinical features, management,pregnancy outcome and prognosis.Results ( 1 ) Etiology:12 cases with obstetric mirror syndrome included 9 cases of Bart's hydrops fetalis,2 cases with fetal complicated congenital cardiac anomalies,and 1 case of unknown etiology.(2)Gestational age at diagnosis and at delivery:gestational age at diagnosis ranged from 28 to 36 weeks [ mean (31.5 ±4.7) weeks],and gestational age at delivery ranged from 28+3 to 38 weeks [ mean (32.9 ±2.9)weeks].There were no significant differences between the gestational age at diagnosis and at delivery in consistented with severe preeclampsia group and mild preeclampsia group [ (31.8 ± 2.3 ) weeks vs.(30.9 ± 7.2) weeks,(32.5 ± 2.3 ) weeks vs.(33.5 ± 3.9 ) weeks,P ＞ 0.05 ].( 3 ) The patients with obstetric mirror syndrome can present a preeclampsia-like syndrome:maternal extremity edema in 12 cases,headache and visual disturbance in 1 case,proteinuria in 11 cases,elevated blood pressure in 5 cases,elevated uric acid in 9 cases,hypoproteinemia in 12 cases,elevated creatinine in 3 case,elevated liver enzyme in 1 case,thrombocytopenia in 2 cases.The major complications included 1 case of HELLP syndrome,acute pulmonary edema,placental abruption,amnionic fluid embolism,DIC respectively,3 cases of acute kidney failure and 6 cases of postpartum hemorrhage.(4) Sonographic findings:① Hydrops fetalis:fetal ultrasound revealed pleural fluid,fetal ascites,skin edema,scalp edema,encephalocolele enlargement, hydropericardium and increased cardio-chest ratio.②Placenta megaly:the placental pathological examination revealed edematous and large in 12 cases.Placental thickness was beyond 4 cm in all cases [ (6.3 ± 1.9) cm ].③Hydramnios:hydramnios could be found in 11 cases [ amniotic fluid index ( 19.7 ± 3.1 ) cm ]. (5) Postnatal conditions:all blood pressure and laboratory findings including urine protein normalized within 5 to 7 days after delivery. (6) Pregnancy outcome:all 12 patients survived,however the perinatal mortality rate was 100％.Two of 12 cases with mirror syndrome underwent cesarean section,and 10 were vaginal delivery,of which 1 need uterine artery embolisom due to postpartum hemorrhage.Conclusions Obstetric mirror syndrome seems to simulate preeclampsia although there are distinguishing features,such as hemodilution,placental edema,and polyhydramnios.When the specific cause of obstetric mirror syndrome can not be identified and corrected,the decision for delivery should be made as soon as possibly.
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Objective To analyze and compare the emergency therapeutic methods for renal colic,and to explore the preferable way of emergency therapies for renal colic.Methods A total of 340 cases of renal colic in emergency department were randomly divided into groups A (pethidine),B (diclofenac sodium),C (phloroglucinol),and D (diclofenac sodium + phloroglucino).The average time of therapeutic effect after given 30 min in different groups was observed.Results The average time of effect in group A [(9.53 ±3.34) min],group D[(10.49 ±2.54) min],and group B[(10.51 ±2.27)min] and they were much shorter than group C [(18.51 ±4.86) min] (P ＜0.01).There was no significant difference in the effective rate among the 4 groups(P ＞ 0.05).The relapse rate of group C (11.43％) and group D (9.00％) were much lower than group A (24.59％) and group B(23.75％).There was no adverse reaction in group C.The rates of adverse reactions in group B (2.27％) and group D (2.78％) were much lower than group A (27.27％).Conclusions Diclofenac sodium combined with phloroglucinol was preferable emergency treatment for renal colic.
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Objective To investigate the risk factors of incidence of premature rupture of membranes and provide reference for prevention of premature rupture of membranes.Methods 890 cases of infants and maternal were studied,analyzing and screening the risk factors of premature rupture of membranes by Logistic.Results The single-factor analysis showed that the pregnant women with diabetes,pregnancy-induced hypertension,multiple preg-nancy and vaginifis had certain degree of contact with incidence of premature rupture of membranes.Multivariate Lo-gistic regression analysis showed that the multiple pregnancy and vaginitis were risk factors of incidence of premature rupture of membranes.Conclusion The multiple pregnancy and vaginitis were risk factors of incidence of premature rapture of membranes,assessment and control these risk factors could prevent and reduce the incidence of premature rupture of membranes.
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Objective To clone NT-3 gene from normal rat brain and to purify its fusion protein and to prepare specific high titer antibody so that to provide a foundation for further study for peripheral nerve injury.MethodsWe amplified target gene by RT-PCR and cloned it into the vector of pMD-18T,then analyzed its sequence and compared it with the sequence from GenBank.We subcloned it into pRSET-A vector and introduced it into Escherichia coli BL21.The expression was induced by IPTG,and identified by SDS-PAGE.The fusion protein was purified by niccolum purify kit.We immuned rabbits with immunological adjuvant for specificity antibody preparation.Results We got a 777 bp gene segment by RT-PCR.The DNA sequence was identical to rat NT-3 gene sequence in GenBank.It proved that the target gene was correctly inserted into the vector.A new protein band of about 34 ku appeared on SDS-PAGE after induction of IPTG.A specific high titer antibody of 1∶64000 was gained by immunizing rabbits with adjuvant.
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【Objective】To observe the in-vitro effect of panax notoginseng saponins(PNS) on bone marrow mesenchymal stem cells(MSCs) proliferation and on inducing MSCs to differentiate into cardiomyogenic cells.【Methods】MSCs were isolated from adult SD rats,and then were cultured and cloned by density gradient method and adhesive cultivation.The cell surface markers of MSCs were detected with flow cytometer.The effect of PNS on MSCs proliferation was observed with methyl thiazolyl tetrazolium(MTT) assay.The 8th generation of continuous cultured MSCs was used to observe the in-vitro differentiation of cardiomyogenic cells.The cardiomyogenic cells were identified under phase contrast microscope by immunohistochemical method and reverse transcription-polymerase chain reaction(RT-PCR) analysis.【Results】CD44 expression of MSCs was positive while that of CD34 expression of bone marrow hematopoietic stem cells was negative,indicating the success of the MSCs culture.After induction with PNS,the increase of MSCs was obvious as compared with the blank control group,and there existed histological changes of MSCs after in-vitro induction.The expression of specific proteins of Desmin and cardiac troponin I(cTnI) in MSCs was positive,and the results of RT-PCR analysis showed that the expression of myosin heavy chain(MHC) was increased.【Conclusion】PNS can increase the in-vitro proliferation of MSCs and induce MSCs to differentiate into cardiomyogenic cells in vitro,and this will supply experimental evidence of cell transplantation for the treatment of myocardial infarction.
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OBJECTIVE:To study the distribution of dosage form of drugs used for outpatients in a general hospital.METHODS:The general information of the outpatients in a general hospital,the total number of the varieties of the prescription drugs as well as the average number for each visit,the rate of using injection for every one hundred patients,and distribution of form of prescription drugs etc were analyzed statistically.RESULTS:Leading the first 5 places on the list of distribution of dosage forms of drugs used for outpatients were tablet,injection,capsule,granule and solution for oral use.Of the total varieties of prescription drugs,drugs taken orally,drugs for injection and drugs for external use stood at 81.07%,13.05%,and 5.88% respectively.CONCLUSION:The use of new variety of drugs such as specific tablets and capsules was little for outpatients in the general hospitals,which deserve to be widely used.