Objective To evaluate the immune responses and its dynamic changes of the babies born to hepatitis B surface antigen (HBsAg) positive mothers after combined passive immunoprophylaxis and active immunoprophylaxis. Methods Two hundred and forty-nine infants born to HBsAg positive mothers were enrolled. All of these infants have received both passive immunoprophylaxis by injecting hepatitis B immunoglobuin (HBIG) and active immunoprophylaxis by vaccinated with hepatitis B vaccine simultaneously 12 hours after birth. After that, all infantscompleted the whole vaccination program. The titers of serum HBsAg and hepatitis B surface antibody (HBsAb) of the infants were checked at 7, 12, 24 and 36 months after birth. The data was analyzed by chi square test. Results Infants born to HBsAg positive mothers showed various immune response modes. The no response rate, low response rate and strong response rate were 8.0% (20/249),11.7% (29/249) and 80.3% (200/249) respectively in the 7-month infants, which were 10.8% (12/120), 26.7% (32/120) and 62.5% (75/120) respectively in 12-month infants. The results from further follow-up showed that no response rate, low response rate and strong response rate were 14.8% (4/27), 33.3% (9/27) and 51.9% (14/27) respectively in the 24-month babies and were 14.3 (1/7), 28. 6% (2/7) and 57.1% (4/7) respectively in the 36-month babies. There were statistically significant difference between the 7-month infants group and other groups (x2= 21.98,P＜0.01). The HBsAb titers of high-response infants group declined over time. The infants with higher antibody titers tended to not decline or decline more slowly. In infants who have even achieved HBsAb titers higher than 1000 mIU/mL, 57.6% (19/33) of them showed decreased titers in 36 months. The titer decrease peaked at 24 month after birth (57.9%, 11/19). In infants who have achieved HBsAb titers of 100 to 1000 mIU/mL, 73.8% (31/42) of them showed decreased titers in 36 months. The titer decrease peaked at 12 month after birth (54.8%, 17/31). HBsAg positive infants usually showed no response at 7 month, who accounted for 70% (14/20,x2 = 128.61, P＜0.01) of all no response infants. Ninety-nine percent (189/191) of HBsAg negative infants showed strong responses. The infants born to both HBsAg positive and hepatitis B e antigen (HBeAg)positive mothers tended to show no response. However, the difference between these infants and others was not statistically significant (9.1% vs 5.5%,x2 =0.24,P＞0.05). Conclusions The immune responses of infants born to HBsAg positive mothers after combined passive and active immunoprophylaxis change over time. The non-response status is usually found in HBsAg positive infants. HBsAg negative infants usually show strong response. Infants born to both HBsAg positive and HBeAg positive mothers tend to show low response. It is recommended to follow standard immunoprophylaxis procedure. The follow-up and active monitor are very important during 7 months to 2 years after birth.

Objective To establish the expression of membrane-bound HLA-G1-G4 isoforms in choriocarcinoma cell line JAR and to investigate its roles in NK cytotoxicity in vitro. Methods Stable expression of HLA-G1, -G2, -G3 and -G4 in JAR cells was established by gene cloning and transfection.HLA-Gtranscripts and protein expression in the transfected JAR cells was tested by RT-PCR, flow cytometry, Western blot and immunocytochemistry, respectively. High-affinity peptide KIPAQFYIL pulsing was performed to evaluate its effects on HLA-G expression. Effects of HLA-G1-G4 isoforms on NK cytotoxicity was performed with lactic dehydrogenase (LDH) releasing method. Results RT-PCR, Western blot and immunocytochemistry results showed that exogenous HLA-G1-G4 gene were successfully transfected and proteins were stably expressed in the HLA-G negative JAR cells; Flow cytometry data showed that only HLAG1, but not HLA-G2-G4 isoform was detectable in those transfected JAR cells and the peptide pulsing did not affect their expression status. However, all HLA-G1-G4 isoform expressed JAR cells could significantly decreased the NK cell cytotoxicity (P＜0.05). Conclusion HLA-G1-G4 isoform expression could dramatically inhibit NK-92 cell lysis, indicating that membrane-bound HLA-G isoforms are importantly immunotolerant and may play immune regulation roles in various physio-pathological situations.

Objective To explore the serological variations for syphilis in infants delivered by treated syphilitic mothers and its influencing factors. Methods Totally, 146 singleton gravidas, who had been treated for syphilis during pregrancy from January 2006 to January 2008 in our hospital, were chosen. Rapid plasma reagin(RPR) and treponema pallidum particle agglutination assay (TPPA) of these mothers before delivery and of the newborns within 3 d after delivery were tested and 92 of the 146 babies were followed up until the age of 24 months. Results (1) Among the 146 neonates, 104 (71.2%) were positive for both RPR and TPPA and 140 (95.9%) TPPA positive only. The RPR positive rate in neonates born to RPR+ + TPPA+ mothers were higher than those born to TPPA+ (only) mothers (81.4% vs 36.4%,χ2 = 25. 3, P<0. 01). 90.4% of the RPR+ neonates (94/104) showed lower or equivalent RPR titers compared to their mothers. (2) Among the 92 babies bein g followed up, the seroreversion of RPR were found in 98. 2%(n = 56) of the 57 babies, who were RPR+ +TPPA+ at delivery, at the 6 months and 100% (n=57) within 8 months, with the peak time within 2 months after birth (78. 9%, n = 45). While, 100% of the babies were found to be TPPA-within 24 mo with the peak time at 10～18 mo (64. 9%, n = 37). For those babies with TPPA+ at delivery, all turned to be TPPA- at 18 mo, with the peak time at 6 ～ 12 mo (57. 1%, n = 20). (3) The seroreversion time of babies with maternal RPR between 1:1～1:4 was later than those with maternal RPR (P<0.05). The seroreversion time of babies with maternal RPR titer of 1:4 was longer than those with maternal RPR titer of 1 > 1 [(2.5±0.8) mo vs (1. 2±0. 4) mo,P<0. 01]. However, the maternal RPR titer did not affect the TPPA reversion time (P > 0.05). The seroreversion time of RPR in infants with neonatal RPR titer of 1 : 4 was later than those with neonatal RPR titer of 1:1 [(3.7±0. 9) mo vs (2. 3±0. 6) mo,P<0. 01], and babies with RPR titer at 1 : 1 - 1 :4 showed longer duration than those with neonatal RPR- in TPPA seroreversion [(11. 2±2. 8) mo, (12.2±2.9) mo, and (11.0±2.2) mo vs ( 6. 9±2. 1) mo, P< 0.01, respectively]. Conclusions Most infants born to syphilitic mothers are serological positive for syphilis despite of standard maternal treatment during pregnancy. Infants, with higher maternal RPR titer during the pregnancy or at delivery, may persist to be serological positive for syphilis for a longer perieod, but all will turn to negative finally. Long term follow up is recommended for serological positive infants, and the diagnosis of congenital syphilis should be cautious.

Objective To investigate the dynamic changes and the clinical significance of T-cell subsets and serum soluble interleukin-2 receptor (sIL-2R)in neonates with hyperbilirubinemia.Methods Thirty-one neonates with hyperbilirubinemia, admitted to the hospital from Decembr 1,2006 to January 31, 2007, were enrolled and divided into two subgroups: severe jaundice group and mild jaundice group according to the bilirubin level. Thirty-two age-mached healty newborns were as controls(control group Ⅰ). The T-cell subsets and sIL-2R of peripheral venous blood samples from these neonates were measured and compared. Sixteen of these 31 neonates with hyperbilirubinemiawere followed up and another twenty-six age-mached healty newborns were as controls(control group Ⅱ ). The level of serum bilirubin in convalescence of sixteen hyperbilirubinemia neonates and control group Ⅱ were tested and analyzed also. Results The levels of CD3, CD4, CD4/CD8 in the neonates with hyperbilirubinemia were lower compared with those of control group Ⅰ [(54.0±5.1)% vs (62.0±4.7)%, (26.8±5.0)% vs (43.0±4.7)%, 0.8±0.1 vs 1.4±0.2] (P＜0.01), but was higher in convalescence than in peak phase[ (62.4±3.3)% vs (55.1±4.2)%, (43.6±2.5)% vs (26.1±4.4)%, 1.4 ± 0.1 vs 0.8±0.1] (P＜0.01). The peak level of sIL-2R in the hyperbilirubinemia group was (319.4± 185.2) kU/L, higher than that in the convalescence [(129.7±99.3) kU/L] and in the control group Ⅱ [(171.9±102.2) kU/L] (P＜0.01). The serum bilirubin level showed negative correlation with CD4/CD8 ( r = -0.99, P ＜ 0.01 ) and positive correlation with sIL-2R (r=0.95, P＜0.05). The sIL-2R level was negatively correlated with CD4/CD8 (r=-0.92, P＜0.05). Conclusions Neonates, when suffering from hyperbilirubinemia, are immunosuppressed which may recover with the alleviation of jaundice.

Objective To investigate the mechanism of different HLA-G isoform mRNA patterns in different cells alters its cell membrane expression.Methods RT-PCR was used to analyze HLA-G isoform mRNA(HLA-Gl-6)of ovarian cancer cell lines HO-8910,HO-8910PM and OVCAR-3,leukemia cell lines Jurkat,K562,HIJ60,MUTIZ-1,and the chorioeareinoma cell lines JEG-3,JAR.HLA-G between cellular membrane and intracellular expression were analyzed by flow cytometry.Results All HLA-G mRNA isoforms were observed in the positive control cell line JEG-3,but none in the negative control cell line JAR.HLA-G1 isoform mRNA was expressed in HO-8910,HO-8910PM,OVCAR-3,MUTZ-1 and Jurkat cells.HLA-G2 mRNA was not detected in any cell line but JEG-3.HLA-G3 mRNA was found in HO-8910,HO-8910PM,K562,HIJ60,MUTZ-1,OVCAR-3 and Jurkat cells.HO-8910,HO-8910PM,HIJ60,Jurkat cells expre8sed HLA-G4 mRNA.Only the Jurkat cells expressed HLA-G5 mRNA.FACS results showed that JEG3 and HO-8910PM cells membrane expressed HLA-G,however,the intraeellular HLA-G expression was detected in all tested cells except the negative control cell JAR.Conclusion Only the HLA-G1 isoform could be exDressed on cell membrane in particular cell lines. Other isoforms including HLA-C2,-G3,-G4,-G5 and HLA-G6 could not reach cell snrface.

Human leukocyte antigen G ( HLA-G) is a member of the non-classical HLA classⅠb family. It is considered to play a crucial role in immune tolerance. A unique feature of HLA-G is the struc-tural diversity as surface expressed and as secreted molecules, which is mainly attributed to alternative spli-cing of the primary transcript. HLA-G can promote the invasion and metastasis of tumor cells through various ways. In addition, HLA-G has been described included in exosomes. Exosomes released by most cell types are nano-sized vesicular bodies that contain lipid bilayer and rich contents. As a new marker for diseases, exosomes are extensively involved in the occurrence and development of diseases. Recent studies have found that exosomes can express soluble HLA-G, which reveal a new way by which HLA-G regulates tumor micro-environment. In this review, we focus on the expression of HLA-G on exosomes to provide new thoughts for the early detection and treatment of tumors.

Objective To probe into the relationship of the cumulative effect of childhood trauma types and symptoms of depression and anxiety among pregnancy women. Methods A total of 276 cases of pregnancy women were investigated by using Childhood Trauma Questionnaire (CTQ- SF), the Edinburgh Postnatal Depression Scale (EPDS) and the State-Trait Anxiety Inventory (STAI). Results The cumulative number of childhood trauma types were positively correlated with the scores of EPDS, state anxiety and trait anxiety (r=0.245, 0.262 and 0.292, P<0.01);the scores of CTQ-SF, EPDS, state anxiety and trait anxiety of multi-CTQ group were higher than that of non-CTQ group, as well as the positive rate of depression symptom and anxiety symptom (P<0.05 or 0.01);Logistic regression analysis showed that the cumulative number and accumulation group of CTQ may be predictors or risk factors of depression and anxiety of women during pregnancy, and existed cumulative effect. When the cumulative number of childhood trauma types more than two types, it increased 2.37 and 3.12 times likelihood of depression and state anxiety comparing to non-CTQ group. Conclusions It suggested that childhood trauma experience may be a risk factors of depression and anxiety during pregnancy, and exist cumulative effect.

Objective To investigate the genetic polym orphism s of 21 autosom al STR loci of Fujian H an population and evaluate the forensic application value of GlobalFilerTM E xpress kit. Methods A m plified w ith GlobalFilerTM E xpress kit, DNA sam ples were obtained from 741 unrelated individuals of Fujian H an population. The population genetics param eters of 21 autosom al STR loci were calculated. Results The 21 autosom al STR loci were found to be no deviation from Hardy-Weinberg equilibration (P>0.05) and relatively abundant in high polym orphism . H eterozygosity ranged from 0.589 to 0.914, pow er of dis-crim ination ranged from 0.754 to 0.992, polym orphic inform ation content ranged from 0.520 to 0.940, and pow er of exclusion ranged from 0.278 to 0.825. The SE33 locus was the highest degree in poly-m orphism . Conclusion The 21 STR loci of GlobalFilerTM E xpress kit have high value in discrim ination pow er and can be useful in personal identification and paternity test in Fujian H an population.

AIM: To investigate the function and morphological changes of long-term cultured primary rat hepatocytes. METHODS: Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method, and then were purified by density and grade centrifugal method with Percoll. Cell viability was observed by 0.4% trypan blue. The hepatocytes were seeded into 6 wells plate with HepatoZYME-SFM medium. ALT, AST, albumin and urea levels in the supernatant were measured, CYPⅠA1 was detected with EROD method. RESULTS: (2-3)?108 cells per whole liver were obtained with viability and purity above 90% after purified with Percoll. Hepatocytes cultured in HepatoZYME-SFM grew well with normal hepatocyte morphometrics. ALT, AST levels in the supernatant decreased after 3-day culture, and kept at a stable level after 6-9 days. Albumin secretion and urea synthesis were maintained at high levels in 18 days, while CYPⅠA 1 enzyme activity was only detected in 3-6 days. CONCLUSIONS: Percoll was used to increase the viability and purity of freshly isolated rat hepatocytes. Hepatocyte morphometrics and their biological synthesized function are effectively maintained in HepatoZYME-SFM medium.

AIM: To Screen and identify human single-chain variable fragment (ScFv) specific to hepatitis B virus core protein and determine its gene sequence. METHODS: The recombinant phages were panned by HBcAg coated in a 96-pore plate and 48 clones were identified specific to HBc after three rounds of panning. The specificity of ScFv from the positive clone was determined by ELISA. Then, the soluble ScFv was expressed in E.coli. HB2151 and secreted in the supernatant. Subsequently, SDS-PAGE and dot blot were performed to identify the ScFv in the supernatant and cell lysate. The gene of ScFv specific to hepatitis B virus core protein was sequenced. RESULTS: The ScFv screened from phage antibodies has a specific combination character with hepatitis B virus core antigen. Soluble ScFv was confirmed to express in E.coli. HB2151 and secrete in the supernatant. The sequence of ScFv gene conformed to that of heavy chain and kappa chain of human immunoglubulin. CONCLUSION: Human ScFv specific to hepatitis B virus core protein has been identified by means of the phage display technology, and its gene sequence has been determined.