Objective To observe the transport of hepatitis B virus (HBV)-infected peripheral blood mononuclear cells (PBMC)through placental barrier set up by choriocarcinoma trophoblast cells (Bewo cells),and to explore the biological role of PBMC as a carrier for HBV transport.Methods Bewo cells and PBMC were cultured and their proliferation and activity were detected by cell counting kit (CCK)-8.One hundred μL serum containing 5 ×10 6 copy/mL HBV DNA was used to infect PBMC,and cells infected with HBV were labeled by fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE).A co-culture model of Bewo cells and HBV-infected PBMC was set up by transwell chamber. The migration of HBV-infected PBMC was detected by flow cytometry.Realtime fluorescence quantitative polymerase chain reaction method was used to detect HBV DNA contents of PBMC under transwell chamber.Results PBMC and Bewo cells proliferated at around 24 h and entered into growth stagnation at around 120 h.The contents of PBMC labeled by green fluorescent at 0,12,24 and 48 h during co-culture under chamber were (0.445 ±0.021)%,(21 .180 ± 4.653 )%,(34.830 ± 7.156 )% and (64.185 ± 3.161)%,respectively.The amount of PBMC marked green fluorescence increased over prolonged incubation time (F =68.983,P =0.001 ).PBMC HBV DNA contents at 24 and 48 h of co-culture under chamber were (1.925±0.431)×103 copy/mL and (2.565 ±0.361)×103 copy/mL,respectively,indicating that PBMC under chamber were infected with HBV.Conclusions PBMC may be a target for HBV infection in extrahepatic tissues.Placental trophoblastic barrier built by transwell chambers may provide new ideas to investigate HBV transmission across the placenta in vitro .

BACKGROUND:The micro-ecological environment has been broken when the ocular prosthesis was inset into the conjunctival sac. The recede of self cleaning function is more conducive to the microbial growth and colonization. The cleaning of plaque biofilm on ocular prosthesis surface affects the patient's wearing comfort and quality of life. It is necessary to seek an effective cleaning method. OBJECTIVE:To compare the clearance effect of five cleaning methods on the palque biofilm on ocular prosthesis surface. METHODS: The conjunctival secretions from 84 patients who were subjected to ocular prosthesis repair were taken for bacterial culture and identification. Fifty pieces of self-curing resin and thermosetting resin artificial eyes were produced. The artificial eyes in each group were randomly divided into five groups, and were cleaned respectively with clear water, volume fraction of 75% ethanol, Boston SIMPLUS, polident and toothpaste. After the completion of the cleaning, the test piece was conducted residual biofilm culture. The clearance effects of different processing modes were evaluated using colony counting method. RESULTS AND CONCLUSION: Eighty-four specimens were submitted for inspection, of which 49 were positive. The Staphylococcusaureus separation rate was 14.29%.Staphylococcus epidermidis separation rate accounted for 13.10%. Maxwel Corynebacterium separation rate accounted for 7.14%. When water, Boston SIMPLUS and toothpaste were used for cleaning, theStaphylococcus aureus colony number in the self-curing resin group was higher than that in the thermosetting resin group (P< 0.05); when ethanol and polident were used for cleaning, there was no difference in the Staphylococcus aureus colony number between these two groups. In self-curing resin, the colony count in the clear water treatment group was higher than that in the other treatment groups (P < 0.05). The colony count in the ethanol treatment group was lower than that in the Boston SIMPLUS group (P < 0.05). There was no significant difference in the colony count between other groups. In thermosetting resin, the colony count in the clear water treatment group was higher than that in the other treatment groups (P < 0.05). There was no significant difference in the colony count between other groups. These results demonstrate that ethanol, Boston SIMPLUS, polident and toothpaste have better cleaning effects onStaphylococcus aureusbiofilms on the surface of two kinds of ocular prostheses than the clear water rinse. Overal, it is encouraged to clean the artificial eyes using polident and Boston SIMPLUS, in order to avoid the occurrence of microbial infection in the conjunctival sac after wearing ocular prosthesis.

Objective To investigate the expression of procalcitonin (PCT) and C-reactive protein (CRP),and endotoxin in bloodstream infection with different microorganisms,so as to assess the value of these inflammatory cytokines in early diagnosis of sepsis in bloodstream infections patients.Methods Data of 152 septic bloodstream infected patients with 90 male and 62 female aged from 62 to 102 years and 79.2 ± 16.3 years in average admitted from January 2012 to December 2013 were analyzed retrospectively.According to the results of blood culture,the microorganisms could be categorized into gram-negative bacteria,gram-positive bacteria and fungus groups,and the levels of serum CRP,PCT,and endotoxin were compared among these groups of bloodstream infections patients within 24 hours after admission.Results (1) A total of 152 strains of microorganisms were surveyed including 92 gram-negative strains (61.18％),43 gram-positive strains (28.29％),and 16 fungal strains (10.53％).In the gram-negative strains,Klebsiella pneumoniae (n =29),Acinetobacter baumannii (n =24),Escherichia coli (n =23),Burkholderia cepacia (n =9) and Pseudomonas aeruginosa (n =4) were the most common isolates.In the Gram-positive strains,13 strains of Staphylococcus aureus were isolated.(2) In the gram-negative bacterial bloodstream infections group,there were 60 (64.52％) patients with endotoxin positive,and there were no endotoxin positive cases with detected gram-positive bacteria and fungal bloodstream infections.The median levels of PCT were significantly different among the three groups [gram-negative strains group:7.760 (3.365,28.585) ng/mL,gram-positive strains group:0.705 (0.265,3.225) ng/mL,fungal infection group:1.245 (0.543,1.998) ng/mL].In the fungal bloodstream infection group,the mean level of CRP was higher than that in other two groups [gram-negative strains group:(126.01 ± 66.53) mg/L,gram-positive strains group:(77.58 ±54.21) mg/L,fungal infection group:(140.14 ±71.21) mg/L].(3) The receiver operating characteristic (ROC) curve of inflammatory cytokines was made for the diagnostic value in bloodstream infections.ROC curve used to distinguish between gram-positive bacterial bloodstream infections group and fungal group showed that AUCPCT+CRP =0.791.When PCT cut-off value was 0.92 ng/mL,and CRP cut-off value was 68.00 mg/L,the sensitivity was 50％ and the specificity was 95.5％.ROC curve used to distinguish between the gram-negative bacterial bloodstream infections group and fungal group showed that AUCPCT+CRP+LPS =0.947.When PCT cut-off value was 2.16 ng/mL and CRP cutoff value was 94.10 mg/L,and endotoxin was positive,the sensitivity was 82.8％ and the specificity was 100％.ROC curve used to distinguish between gram-negative bacterial bloodstream infections group and gram-positive bacterial group showed that AUCPCT+CRP+LPS =0.947.When PCT cut-off value was 2.68 ng/ mL,CRP cut-off value was 106.5 mg/L,endotoxin was positive,the sensitivity was 74.2％ and the specificity was 97.7％.Conclusions Gram-negative bacteria were the most common microorganisms in bloodstream infections in ICU patients.Compared with single inflammatory cytokine,the serum concentrations of PCT,CRP and endotoxin used together could provide more sensitivity and specificity for the early diagnosis of bloodstream infection with different microorganisms.

Objective To investigate how surgical patients are assessed regarding pressure ulcer risks and the usage of evaluation tools in different stages during perioperative period.Methods Totally 210 nurses from operating rooms and surgical wards in 7 tertiary hospitals of Jiangsu Province were investigated with self-designed questionnaire.Results Operating room nurses of 7 hospitals used different kinds of scales for pressure ulcer risk assessment,the same scale document was used during the different stages of operation,the frequency and time for pressure ulcer risk assessment varied from one to three times,preoperative visit to handover in surgical ward were different;the Braden scale was used by surgical nurses to evaluate pressure ulcer risk after operation,while most surgical nurses believed that the existing scale lacked specificity and intraoperative risk factors that should be considered when assessing postoperative patients.Conclusion The frequency and time of pressure ulcer risk assessment of patients during perioperative period are not standardized,the content of scale is limited and lacks specificity,the assessment of pressure ulcer risk during perioperative period is discontinuous and non-dynamic.There are no specific risk assessment scales targeting preoperative,intraoperative and postoperative pressure ulcer risk assessment.

Objective To evaluate the effect of combination therapy with milrinone and esmolol on hemodynamics and cardiac function in patients with septic myocardial depression. Methods From October 2010 to October 2013,after the hemodynamics and cardiac function were evaluated by pulse indicator continuous cardiac output (PICCO),74 sepsispatients withCI < 2.2 L/min · m2 after fluid resuscitation were enrolled in the study and were divided into group A with intravenous injection of dobutamine hydrochloride ,and group B with intravenous injection of milrinone and esmolol,with 37 cases in each group. The patients'PICCO indicators, echocardiography and cardiac biomarker(CK,CK-MB,MYO,cTnI and ProBNP)in two groups were compared before and after 3-day treatment. Results (1)CI and GEF were significantly increased in group B after 3-day treatment when compared with those in group A.(2)Compared with those in group A,early diastolic mitral flow velocity/end diastolic mitral velocity (E/A) and right ventricular diastolic diameter(RVD) in group B had statistical significance.(3) CK-MB,cTnI and ProBNP decreased significantly in group B when compared with those in group A. Conclusion Combination therapy with milrinone and esmolol can increase cardiac ejection function,slow down the heart rate,reduce the heart blood and vascular preload,lessen the injury of myocardial and improve heart function.

Objective To investigate the effects of volatile oil from artemisia dracunculus on myocardial injury caused by viral myocarditis in mice and explore its possible mechanism.Methods Totally 160 adult male BALB/c mice were randomly divided into normal control group (10) and viral myocarditis group (150).Viral myocarditis mice models were reproduced by intraperitoneal inoculation with a solution of coxsackievirus B3 (CVB3),a viral strain with affinity to myocardium,and then randomly divided into model,astragalus group,and low-,medium-,and high-dose volatile oil from artemisia dracunculus groups.After 1 hour of viral infection,normal control group and model group mice were given normal saline by intragastric administration,astragalus group mice were injected with astragalus 0.1 mL in each mouse by intraperitoneal injection,and the mice in other three groups were given low,medium and high dose (2％,5％,10％) 0.3 mL volatile oil from artemisia dracunculus in each mouse by intragastric administration,respectively,once a day for one week consecutively.The mortality,heart/body weight ratio,the activity of natural killer cells (NK cell),virus titer in myocardial homogenate,serum cardiac troponin Ⅰ (cTnI) level and myocardial pathological changes were observed.Results ① Mortality:the mortality of model group was higher than that of the normal control group,astragalus group,low and medium dose volatile oil from artemisia dracunculus groups (60.0％ vs.0％,23.3％,20.0％,28.7％),and the difference in the mortality being of no statistical significance between model group and that of high-dose volatile oil from artemisia dracunculus group (60.0％ vs.47.6％,P ＞ 0.05);the mortality of astragalus group was obviously lower than that of high-dose volatile oil from artemisia dracunculus group (P ＜ 0.01),and the differences in comparisons between the mortalities of astragalus intervention group,and medium-and low-dose volatile oil groups were not statistically significant (all P ＞ 0.05),and the comparison of mortality between low-and medium-dose volatile oil groups were also not statistically significant (P ＞ 0.05).② Immunization parameters:on the 8th day after modeling,the activity of NK cells in the model group was significantly lower than that in the normal control group [(15.91 ± 3.87)％ vs.(38.50 ± 2.32)％],the activities of NK cells in astragalus group,medium-and low-dose volatile oil from artemisia dracunculus groups were significantly higher than that in model group [(19.38 ± 3.27)％,(18.54 ± 3.09)％,(18.36 ± 2.64)％ vs.(15.91 ± 3.87)％,all P ＜ 0.05].None of virus was detected in the myocardial homogenate in the normal control group,and the virus titers in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than the titer of the model group (10-9/mL:1.96 ± 0.44,1.95 ± 0.46,1.95 ± 0.48 vs.2.41 ± 0.51,all P ＜0.01).③ Myocardial injury parameters:the level of cTnI in the normal control group was less than 0.1 μg/L,obviously lower than that in the model group [(15.84 ± 3.89) μg/L],as well as the ratio of heart/body weight in model group was also significantly higher than that in normal control group (× 10-4:8.3 ± 1.3 vs.4.6 ± 0.1),and the cTnI and the ratio of heart/body weight of astragalus intervention group,low and medium dose volatile oil from artemisia dracunculus groups were markedly lower than those of model group [cTnI (mg/L):10.03 ± 2.35,10.81 ± 2.56,11.10 ± 1.89 vs.15.84 ± 3.89,ratio of heart/body weight (× 10-4):7.2 ± 0.8,7.3 ± 1.0,7.3 ± 0.6 vs.8.3 ± 1.3].In the normal control group,there were no inflammatory cell infiltration and necrosis in myocardial tissue,the scores of myocardial pathological changes were 0.In the model group,the scores of inflammatory cell infiltration (3.25 ± 0.45) and of necrosis (2.91 ± 0.51) were markedly higher than those in the normal control group.And the above scores in astragalus group,low and medium dose volatile oil from artemisia dracunculus groups were significantly lower than those of the model group (infiltration score:2.92 ± 0.39,2.95 ± 0.35,2.95 ± 0.37 vs.3.25 ± 0.45,necrosis score:2.46 ± 0.50,2.50 ± 0.51,2.54 ± 0.50 vs.2.91 ± 0.51,all P ＜0.05).Conclusions Volatile oil from artemisia dracunculus can protect cardiomyocytes by removing the virus and regulating the immune function in the body.But the protective effects of volatile oil from artemisia dracunculus is related to the dosage,and the effects of low and medium dose are better.

Objective To explore the curative effect of 308 nm excimer laser on child vitiligo and nursing measures. Method Fourty-eight children with vitiligo were treated with 308 nm excimer laser and nursed with targeted psychological treatment, skin and diet nursing.Result After treatment,6 of them got recovered(12.5%),24 remarkably improved(50%),15 slightly improved(31.2%)and 3 ineffectively treated(6.2%).Conclusions 308 nm excimer laser is effective for the treatment of child vitiligo.High quality nursing can ensure and improve the efficiency of the 308 nm excimer laser.

ObjectiveTo observe the role of Sijunzi decoction on the gut barrier protection and immunity regulation in spleen-qi deficiency type patients with persistent inflammation-immunosuppression catabolism syndrome (PICS).Methods A prospective study was conducted, and according to random number table, 46 patients with PICS accompanied by spleen-qi deficiency admitted to Department of Critical Care Medicine of Beijing Shijitan Hospital of Capital Medical University were randomly divided into two groups: control group and Sijunzi decoction group(each 23 cases). Conventional therapy was given to both groups, and the patients in Sijunzi decoction group were additionally treated with modified Sijunzi decoction 100 mL by nasal feeding, while those in the control group were treated with an equal amount of warm boiled water by nasal feeding. The course of treatment was 2 weeks in both groups. The improvement in traditional Chinese medicine(TCM) syndrome and gastrointestinal function was observed in two groups before and after treatment. At the same time, the changes of lymphocyte subsets including total T cells(CD3+ cells),helper/inducer T cells(CD3+/CD4+T cells), suppressor/cytotoxic T cells(CD3+/CD8+ T cells), CD4/CD8, total natural killer cells(NK cell,CD3-/CD16+CD56+ cells),natural killer like T cells(NK T cell,CD3+/CD16+CD56+ T cells),total B cells(CD19+ cells),regulatory T cells(Treg T cells, CD4+/CD25+ T cells),suppressor T cells(CD8+/CD28- T cells) and cytotoxic T cells(CD8+/CD28+ T cells)were analyzed.Results① There were no differences in the acute physiology and chronic health evaluationⅡ(APACHEⅡ)score,acute gastrointestinal injury(AGI)grades and peripheral blood lymphocyte subsets between Sijunzi decoction group and control group before treatment( allP>0.05).② After 2 weeks of treatment, all the patients' clinical symptoms of spleen-qi deficiencywere improved, and there was no difference in effect between the Sijunzi decoction and control group(valid:11 cases vs. 6 cases, invalid:12 cases vs.17 cases, bothP>0.05) . In both groups, the patients' AGI condition was improved, but the improvement in treatment group was more significant than that in the control group(valid: 14 cases vs. 7 cases, P<0.05) .③ In the control group,the numbers of CD3+, CD3+/CD8+,CD8+/CD28- T cells after treatment were increased significantly〔CD3+:(62.37±7.83)% vs.(54.08±11.65)%, CD3+/CD8+:(31.52±10.55)% vs. (23.94±9.22)%, CD8+/CD28-:(24.97±10.25)% vs.(16.78±10.55)%〕 and CD19+ number was decreased obviously〔(5.78±5.33)% vs.(9.73±8.02)%〕 at the same time(allP<0.05). After treatment, in the Sijunzi decoction group, the numbers of CD3+,CD3+/CD4+, CD3+/CD8+, CD8+/CD28-, CD8+/CD28+ T cells were increased significantly compared with those before treatment〔CD3+:(74.53±7.64)% vs.(52.98±10.05)%, CD3+/CD4+:(36.27±12.08)% vs.(30.00±8.60)%, CD3+/CD8+:(37.33±12.56)% vs.(22.88±9.97)%, CD8+/CD28-:(26.89±10.80)% vs.(17.01±9.48)%, CD8+/CD28+:(12.08±5.50)% vs.(8.47±4.29)%〕, and total CD19+ number was decreased remarkably at the same time〔(4.60±4.28)% vs.(9.86±8.61)%,P<0.05〕. In the Sijunzi decoction group, after 2 weeks of treatment, the number of CD3+ T cells was increased more significantly than that in the control group〔(8.29±9.28)% vs.(5.80±5.33)%,P<0.05〕.ConclusionUsing Sijunzi decoction combined with conventional therapy can improve the clinical symptoms of patients with PICS accompanied by spleen-qi deficiency and gastrointestinal dysfunction, the mechanism is possibly via enhancing immunity by regulating the numbers of B and T lymphocytes, and maybe the cell-mediated immunity plays a more prominent role.

Objective To study the relationship between placenta HBsAg in HBsAg positive pregnant women and serum hepatitis B virus (HBV) markers,HBV DNA levels in newborns.Methods Placenta HBsAg was detected by immunohistochemical affinity hormone-biotin complex (ABC) method in 155 HBsAg positive pregnant women.Serum HBV markers in newborns were detected by enzyme-linked immunosorbent assay (ELISA).Serum HBV DNA levels of newborns were detected by real-time fluorescence quantitative polymerase chain reaction (PCR).The positive rates were compared using x2 test.Results HBsAg was expressed with different levels in various types of cells of placenta in 155 pregnant women.The total placenta HBsAg positive rate was 37.4％ (58/155),and those in decidual cells,trophoblastic cells,villous mesenchymal cells and villous capillary endothelial cells were 37.4％ (58/155),25.8％ (40/155),18.7％ (29/155) and 7.1％ (11/155),respectively.The HBsAg positive rates of placenta gradually decreased from decidual cells of the maternal surface to villous capillary endothelial cells of the fetal surface (tendency x2 =43.01,P=0.00).The positivity of placenta HBsAg was associated with both HBsAg and HBeAg in newborns (x2 =4.88,P＜0.05 and x2 =3.86,P＜0.05,respectively),while that was not associated withanti-HBe and anti-HBc in newborns (x2 =3.36,P＞0.05 and x2 =0.00,P＞ 0.05,respectively).The risk of HBsAg positive in newborns was higher when HBsAg was positive in villous capillary endothelial cells and villous mesenchymal cells (OR=5.31,95 ％CI=1.38-20.40 and OR=3.33,95％CI=1.16-9.52,respectively).The risk of HBeAg positive in newborns was higher when HBsAg was positive in trophoblastic cells and villous mesenchymal cells (OR=3.04,95 ％CI=1.45-6.39 and OR=3.05,95 ％ CI=1.32-7.03,respectively).However,placenta HBsAg positive was not associated with HBV DNA positive in newborns (x2 =0.09,P＞0.05).Conclusion The risk of neonatal HBV serological markers positive is higher when the HBsAg positive placental cells are closer to fetal surface,which indicates that HBsAg enters fetal blood circulation by means of cell transferring layer by layer.

Objective We observed the effect of dexamethasone ointment on preventing phlebitis induced by vinorelbine. Methods Patients with malignant tumor who received chemotherapy of vinorelbine through peripheral superficial vein injection were divided into the observation group (70 cases) and the control group (72 cases) according to the date of hospitalization. All patients received vinorelbine four times averagely. Patients in the observation group was given dexamethasone ointment along punctured superficial vein. Patients in the control group received routine nursing measure. The incidence rate, time and degree of phlebitis was compared between these groups. Results The incidence rate and degree of phlebitis was lower than those of the control group (P< 0.01, P< 0.05). The incidence time of phlebitis in the observation group was also later than that of the control group (P<0.05). Conclusion Local application of dexamethasone ointment could effectively reduce the incidence of superficial phlebitis caused by vinorelbine chemotherapy.