Objective To investigate the molecular epidemiology of hepatitis B virus(HBV) in child carriers. Methods Blood samples were collected from children under 15 in Jiangsu and Zhejiang provinces. Enzyme immunoassay(EIA) and microparticle enzyme immunoassay(MEIA) were applied to screen hepatitis B surface antigen(HBsAg) positive children. Nested-PCR and real time PCR were used to amplify the HBV S gene and detected HBV DNA loads. S gene sequence and three-dimensional structure were analyzed by the DNASTAR and VMD1.8.6, respectively. SPSS 12.0 software was applied for data processing. Results A total of 64 HBsAg-positive cases were found in the screened children, from which 41 HBV S gene sequences were obtained. The average HBV DNA loads were(4.15±0.79)×10~7 copies/mL in 64 HBV carriers. Among 41 sequences. genotype C, B and B+C accounted for 82.93%(34/41), 12.19%(5/41)and 4.88%(2/41), respectively; and the serotypes were adr(34/39,87.18%), adw(4/39,10.24%) and ayr (1/39, 2.56%) with 2 strains unable to be sub-typed. The most common variants of "a" determinant in HBV S gene were 129 site Q→F(glutamine→phenylalanine), 145 site G→R(glycine→lysine), 131 site S→N(serine→asparagine)and 144 site C→A(cysteine→alanine), and the mutation frequencies were 12.20%(5/41), 4.88%(2/41), 2.27%(1/41)and 2.27%(1/41), respectively . The total mutation frequency was 21.95%(9/41). The S protein spatial structures of 129 site "Q→F" and 145 site "G→R" were entirely different from that of the wild strain. Conclusion Wild strain of HBV(C/adr) is predominant in the children HBV carriers, exhibiting a high replication, and the HBV vaccine should be still effective.

Objective An ELISA-based assay for detecting alloantibodies against FⅧ was established to estimate the incidence of alloantibodies against FⅧ in treated patients with hemophilia A. Methods One hundred and forty patients with hemophilia A and 80 healthy controls were enrolled. Among hemophilia A patients, 84, 34 and 22 patients were in severe, moderate and mild conditions respectively. All patients were treated with plasma-derived FⅧ concentrates before. The titer wells were coated with MoAb against FⅧ which was developed in our laboratory. Then human recombinant FⅧ concentrates were applied. After incubation in room temperature for 2 hours, diluted plasma samples and HRP-conjugated goat anti-human IgG were added successively. Finally Absorbance (A490) were measured and recorded. Inhibitor activity against FⅧ for all plasma samples was measured by a modified Nijmegen assay simultaneously. Results The results showed that alloantibodies against FⅧ were found in 40.0% (56/140) patients by ELISA. And the alloantibody incidences in the severe and non-severe patients were 47.6% (40/84) and 28.5% (16/56)respectively. There was statistical significance between these two categories (x2 = 5.079, P ＜ 0.05 ). The FⅧ inhibitor activity was detected in 24.3% (34/140) patients by modified Nijmegen assay. The inhibitor incidences in the severe and non-severe patients were 33.3% (28/84) and 10.7% (6/56) respectively.There was statistical significance (x2 = 9.349, P ＜ 0.05). Twenty-five patients were positive for FⅧ alloantibodies by ELISA but had no FⅧ inhibitor activity by the modified Nijmegen assay. The positive rates of FⅧ alloantibodies and inhibitor activity were 40.0% (56/140) and 24.3% (34/140) respectively,which had significant difference (x2 = 15.75, P ＜ 0.01 ) and strong positive correlation ( rn = 0.59, P ＜0.01 ). Meanwhile the results deduced from these two tests shared a high consistency rate ( Kappa = 0.55,P ＜0.01 ). Conclusions The detection rate for alloantibodies against F Ⅷ is enhanced by our newlydeveloped ELISA. Our results suggest that the occurrence of the alloantibodies against F Ⅷ in Chinese hemophilia A patients is not rare and the alloantibody incidence is preponderant in the patients with severe hemophilia A compared with non-severe hemophilia A patients.

Background and purpose:The previous research has found that the prostate stromal cells derived from different prostate zones have distinct effect on prostate epithelial cells. We also revealed that LMO2 protein was highly expressed in PZ stromal cells (PZSCs) and prostate cancer associated fibroblasts (CAFs) compared with TZ stromal cells. This study investigated the effect of LMO2 protein in prostate stromal cells on proliferation and invasion of prostate cancer PC-3 cells and its mechanisms. Methods:Lentivirus overexpression vectors were used to establish LMO2-overexpressed prostate WPMY-1 stromal cell line. shRNA plasmids were used to suppress LMO2 in CAFs. LMO2 mRNA and protein level of both WPMY-1 and CAFs were evaluated by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Then, PC-3 cells were co-cultured with different prostate stromal cells and the in vitro proliferation and invasion of PC-3 were measured by CCK-8 and matrigel invasion assays respectively. Results:When co-cultured with LMO2-overexpressed prostate stromal cells, both proliferation and in-vasion of PC-3 were improved. However, when co-cultured with CAFs which have inhibited expression of LMO2, the proliferation and invasion of PC-3 were reduced. The protein array proifling found that both interleukin-11 (IL-11) and ifbroblast growth factor-9 (FGF-9) were enhanced extensively in the supernatant collected from LMO2-overexpressed WPMY-1 cells. Conclusion:The expression of LMO2 in prostate stromal cells could be responsible for development of prostate cancer. Paracrine of cytokines, such as IL-11 and FGF-9, from LMO2-overexpressed stromal cells had effects on the proliferation and invasion of prostate cancer cells.

Objective To investigate the effects of B lymphocyte-induced maturation protein-1 ( Blimp-1) on the number and function of splenic lymphocytes.Methods The mice with defective Blimp-1 in T cells were generated by cross-breeding B6.Blimp-1flox/flox mice with B6.Lck-Cre mice.The mononuclear lymphocytes isolated from spleen of T cell conditional Blimp-1 knockout (Blimp-1CKO) mice and wild type ( WT) C57/B6 mice were comparatively analyzed.Alterations of CD4+T and CD8+T cell subsets, the secre-tion of cytokines as well as the expression of C-C chemokine receptor type 7 ( CCR7 ) and Sphingosine-1-phosphate receptor 1 (S1P1) in mice from the two groups were analyzed by flow cytometry.The changes of CD19+B cell subsets were also detected.Results Compared with WT mice, the total numbers of mononu-clear cells, T and B lymphocytes were all significantly increased in Blimp-1CKO mice ( P<0.05) .The ab-solute numbers of CD4+T, CD8+T and CD19+CD5+CD1d+B cells in mice form Blimp-1CKO group were higher than those of the control group (P<0.05), however, no significant differences with the percentages of these cell populations were observed between two groups.Higher numbers and percentages of CD19+CD5+B cells were detected in mice from Blimp-1CKO group (P<0.01).The Blimp-1CKO mice showed increased secretion of IFN-γ, TNF-α, IL-17 and IL-2, but decreased expression of CCR7 on CD8+T cells as com-pared with WT mice (P<0.05).No significant differences with the changes of S1P1 were found between the two groups.Conclusion Blimp-1 played an important role in the maintenance of number, phenotype and function of T cells.Furthermore, not only T cells but also B cell subsets in mice were affected by the dele-tion of Blimp-1 in T cells.

Objective To compare the effectiveness of training with a digital video disc ( DVD)-based instruction ver-sus intubation model simulator in placement of endotracheal tube by video -laryngoscope .Methods Sixty patients un-dergoing elective gynecology surgeries and twenty residents of Dept .of anesthesiology were randomized to place a en-dotracheal tube by video-laryngoscope .Results Compared with intubation model simulator , intubation time [ (68 ± 14)s vs (69 ±7)s], score of performance (8.24 ±0.64 vs 8.31 ±0.58), failure rate (3 times/11 vs 1time/9 resi-dents), and instruction rate (5times/11 vs 4times/9 residents) were no statistic difference .Conclusions Digital video disc ( DVD)-based instruction is an alternative way to intubation model simulator in training .

AIM:To explore efficacy of the different kinds of artificial tears treatment in patients with xerophthalmia after phacoemulsification combined with intraocular lens implantation.METHODS:Totally 280 patients (280 eyes) with xerophthalmia after operation in our hospital from January 2015 to June 2016 were selected.According to the different treatment methods, they were divided into control group (n=70;treated with tobramycin and dexamethasone eye ointment 3 times per day for 1wk, levofloxacin eye drops 3 times per day for 1wk, pranoprofen eye drops 4 times per day for 1mo), polyacrylic acid group (n=70;besides the treatment of control group, polyacrylic acid was used 4 times per for 1mo), polyethylene glycol group(n=70;besides the treatment of control group, polyethylene glycol was used 4 times per for 1mo) and sodium hyaluronate group (n=70;besides the treatment of control group, sodium hyaluronate was used 4 times per for 1mo).The tear film break up time (BUT), Schirmer Ⅰ test (SⅠt), symptoms of dry eye and corneal staining in four groups were observed.RESULTS:(1) BUT:The BUT of the four groups significantly increased after treatment(P<0.05);that of polyacrylic acid group, polyethylene glycol group, and sodium hyaluronate group was different compared with control group(P<0.05);the BUT in sodium hyaluronate group was significantly higher than the other groups after 2wk of treatment(P<0.05).(2) SⅠt:SⅠt of the four groups significant increased after treatment(P<0.05);that of polyacrylic acid group, polyethylene glycol group, and sodium hyaluronate group was different compared with control group(P<0.05);and the SⅠt in sodium hyaluronate group was significantly higher than other groups (P<0.05).(3) Xeroma score:the scores of dry eye significantly decreased after treatment in the four groups(P<0.05);that of polyacrylic acid group, polyethylene glycol group, and sodium hyaluronate group was different compared with control group(P<0.05);and the scores of the sodium hyaluronate group after 3wk was significantly lower compared with other groups(P<0.05).(4) Corneal staining score:the scores significantly decreased after treatment in the four groups(P<0.05);at 1 and 2wk after treatment the corneal staining score had not statistically different among the four groups (P>0.05);sodium hyaluronate group was significant lower than other groups in corneal staining score at 3wk and 1mo after treatment(P<0.05).CONCLUSION:Artificial tears in the treatment of xerophthalmia after cataract phacoemulsification combined with intraocular lens implantation has better clinical efficacy, which contains sodium hyaluronate may be the better than others.

Objective:To compare the patient-centered doctor-patient communication skills in anesthesiologists with different working experience and genders.Methods:The second and third year residents who had been trained as residents in the Department of Anesthesiology in our hospital from September 2017 and September 2018 and senior physicians who had worked for 10-15 yr in our hospital, were recruited to perform a preoperative interview in a standardized patient counter.Consultation and Relational Empathy (CARE) measure was used to assess the communication skills among senior physicians, residents, and anesthesiologists of different genders.The residents also took the objective structured clinical examination (OSCE) and theory examination.Results:Thirty-six residents and 20 senior physicians were included.There was no significant difference in the CARE measure score between residents and senior physicians and in the total CARE measure score between residents who received doctor-patient communication training at the stage of medical students and those who did not ( P>0.05). Compared with the residents in the second year, the listening ability score and decision-making ability score were significantly decreased ( P<0.05), and no significant change was found in the scores for the other items in the residents in the third year ( P>0.05). Compared with residents, the listening ability scores were significantly increased, and the caring ability scores were decreased ( P<0.05), and no significant change was found in the scores for the other items in senior physicians ( P>0.05). There was no difference in CARE measure scores between the male and female anesthesiologists.The resident′s CARE measure score was positively correlated with the annual OSCE score ( r=0.486, P<0.05), and there was no correlation between the resident′s CARE measure score and annual theoretical examination grade ( r=0.308, P>0.05). Conclusion:Senior anesthesiologists with more clinical experience are not superior to the junior residents in patient-centered communication skills, and the doctor-patient communication skills of residents are not related to their medical theoretical grade.Moreover, there is no significant difference between male and female anesthesiologists in terms of doctor-patient communication skills before surgery.Therefore, training of patient-centered doctor-patient communication skills should be strengthened at all levels of anesthesiologists.

Objective: To analyze the clinical and laboratory abnormalities of two patients with α1-antitrypsin (α1-AT) Pittsburgh in a family and review the literatures. Methods: Both plasma clotting time and factor activities were performed using clotting or substrate methods. Platelet aggregation was evaluated using an optical aggregometer. The serum protein electrophoresis was performed on Sebia HYDRASYS by using Agarose gel. The exons of α1-AT were amplified by using polymerase chain reaction (PCR) and then sequenced and compared with NCBI GenBank records. Results: The proband had several ruptures of corpus luteum and bleeding after operation, while her daughter had no bleeding history. Both of them showed prolonged coagulation tests which could not be corrected by mixing with the normal plasma. They also showed low levels of plasma coagulation factors, undetected protein C and S activity and abnormal bands of α1-globulin. The results of gene sequencing demonstrated that they were heterozygous for g.T17132G (p.Met358Arg) mutation of α1-antitrypsin gene (NG_008290.1) . Conclusions Comparing with the data of previously reported cases, our results confirmed the obvious abnormality of coagulation test and the discrepancy of bleeding tendency of α1-antitrypsin Pittsburgh patients, and suggested that the rupture of corpus luteum would be a specific characteristic in women of child-bearing age.

Objective: To evaluate efficacy and safety of retrograde intrarenal stone surgery for treatment of upper urinary calculi. Methods: The clinical data of 640 patients with upper urinary tract calculi treated by retrograde intrarenal stone surgery (RIRS) in Renmin Hospital of Wuhan University from April 2016 to January 2019 were retrospectively analyzed. There were 424 males and 216 females. The awerage age was (46.2±12.8) years old, ranging 18 to 76 years old. The maximum diameter of the stone is (1.4±0.7) cm, ranging 0.6-3.2 cm. There were 126 cases with inferior calculi and 514 cases with non-lurgical calculi. There were 196 cases with unilateral ureteral calculi, 118 unilateral ureteral calculi cases with renal calculi, 236 cases with unilateral renal stones, and 90 cases with double kidney stones. 104 cases were placed with double J tube before operation and 496 cases were not placed before operation. There were 8 cases of horseshoe kidney, 30 cases of isolated kidney with renal insufficiency, 4 cases of pelvic ectopic kidney with dysplasia, 6 cases of congenital ureteral malformation and 2 cases of sponge kidney. Preoperative average hemoglobin was (133.2±5.6)g/L, ranging 126-188 g/L.And average serum creatinine was (84.4±12.2)μmol/L, ranging 74-242μmol/L before operation. All patients were treated with general anesthesia under the lithotomy position. The ureteroscopy combined with holmium laser lithotripsy was performed.The 200μm fiber was used, which the parameters were set as 12-45 W(0.5-1.5 J/10-30 Hz). The stone baskets were used to take stones according to actual conditions. The operation was performed by doctors of the same qualifications. Results: All patients underwent successful operation. The mean operation time was (45.6±14.6)min. The average postoperative hospitalization was (4.8±1.5)d. The postoperative serious complication rate was 0.9%, including(2 cases of sepsis and 1 case of subcapsular hematoma. Of the 640 patients, 596 were admitted to the hospital for a double J tube and 44 were lost of follow-up. 552 patients met the stone removal criteria, 44 patients did not meet the stone removal criteria for other treatments, such as extracorporeal shock wave lithotripsy, ureteroscopy or observed regularly. The stone-free rate (SFR) was 92.6% (552/596) after 1-3 months. On the first postoperative day, serum creatinine was (76.0±10.6)(58-156) μmol/L, and postoperative hemoglobin was (126.4±9. 6)(120-176) g/L. There was no significant difference in preoperative and postoperative hemoglobin (t=2.02, P=0.064). Preoperative and postoperative creatinine (t=64.76, P<0.05) was statistically significant. Meanwhile, the stone size (χ²=29.569, P<0.05) and position (χ²=44.949, P<0.05) versus SFR the impact was statistically significant. Multivariate regression analysis showed that stone size was not an independent risk factor for stone clearance (P=0.639). The stone position was an independent risk factor for stone clearance (P=0.013). Conclusions RIRS is a reliable treatment for small and medium calculi patients of the upper urinary tract. The curative effect of stone removal is clear, the complications are few, the safety is high. However, there are certain limitations to the efficacy in the treatment of large stones and lower calculi. Lower calculi is the independent risk factor for the treatment of efficacy.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree affected with neurofibromatosis type I (NF1). METHODS: Target capture high-throughput sequencing and Sanger sequencing were carried out to detect the pathological variant in a NF1 patient and his parents. RESULTS: The proband and his similarly affected father have both harbored a novel nonsense variant of c.2511G>A (p.trp837x) in the NF1 gene. The same variant was not found in his mother and 200 healthy controls. CONCLUSION The heterozygous nonsense variant of c.2511G>A (p.trp837x) of the NF1 gene probably underlay the pathogenesis of NF1 in this pedigree.
China ; Heterozygote ; Humans ; Mutation ; Neurofibromatosis 1/genetics* ; Pedigree