Objective To detect the antibodies against recombinant chlamydial plasmid-encoded protein 3(rPgp3),chlamydial protease-like activity factor(rCPAF),Ct143 encoded protein(rCT143), Ct101 encoded protein(rCT101),Ct694 encoded protein(rCT694),Ct813 encoded protein(rCT813), Chlamydia membrane protein A(rIncA),Ct875 encoded protein(rCT875),major outer membrane protein (rMOMP)and heat shock protein 60( rHsp60)in serum samples collected form patients with urogenital Chlamydia trachomatis(Ct)infection and to evaluate the antigenicity of those proteins. Methods The re-combinant plasmids expressing the 10 proteins and a blank plasmid were transformed into E. coli BL21 strains,respectively. The transformed E. coli BL21 strains were induced by isopropyl β-D-1-thiogalactopyr-anoside(IPTG)to express recombinant proteins. The glutathione pre-coated 96-well ELISA plates were coa-ted with lysates. Serum samples were collected from 50 patients with Ct infection and 10 patients without Ct infection. ELISA was performed to detect the antibodies against 10 recombinant proteins. Results The anti-bodies against rPgp3,rCPAF,rCT143,rCT101,rCT694,rCT875,rCT813,rMOMP,rIncA and rHsp60 proteins were respectively detected in 44 cases(88% ),38 cases(76% ),37 cases(74% ),36 cases (72% ),33 cases(66% ),31 cases(62% ),30 cases(60% ),26 cases(52% ),24 cases(48% )and 17 cases(34% )out of 50 serum samples. No antibodies against 10 recombinant proteins were detected in the serum samples collected from patients without Ct infection. Conclusion The rPgp3 protein showed the strongest antigenicity among all of the studied proteins,followed by rCPAF and rCT143 proteins. The rHsp60 protein showed the lowest antigenicity.

OBJECTIVE:To observe clinical effects and safety of small dose of octreotide for preventing hyperamylasemia and acute pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).METHODS:One hundred and twenty ERCP pa tients were selected from our hospital during Oct.2014-Jan.2015 and then divided in to observation group and control group in accordance with random number table,with 60 cases in each group.Both groups were given Diazepam tablet 10 mg+Meperidine hydrochloride tablet 100 mg+Phenobarbital scopolamine tablet 2 tablets 0.5 h before surgery for sedation and analgesia,and routine acid suppression and anti-infective therapy.Observation group was additionally given Octreotide acetate injection 0.1 mg hypodermically and then given Octreotide acetate injection 0.1 mg immediately after surgery,8 h after surgery.The levels of serum amylase and blood glucose were observed in 2 groups,and the occurrence of postoperative complication and ADR were recorded.RESULTS:Before operation,there was no statistical significance in the levels of serum amylase and blood glucose between 2 groups (P＞ 0.05).After operation,the level of serum amylase in control group was significantly higher than in observation group,with statistical significance (P＜0.05).There was no statistical significance in blood glucose level between 2 groups after operation (P＞0.05).The incidence of hyperamylasemia and ADR in observation group was significantly lower than in control group,with statistical significance (P＜0.05),and there was no statistical significance in the incidence of acute pancreatitis between 2 groups after operation (P＞0.05).CONCLUSIONS:Small dose of octreotide can effectively reduce the level of serum amylase and the incidence of hyperamylasemia after ERCP with good safety.

20 first mandibular premolars were randomly divided into 2 groups(n =10).The teeth in experimental group were treated by fi-ber main post in combination with auxiliary pile,in control group by single fiber main post,and then were restored by metal crown.They were fixed in universal testing machine.The fracture load(N)of experimental and control group was (846.50 ±40.33)and (437.90 ±41.15) respectively(P <0.01).

AIM: To discuss the effect of wearing customized curvature soft corneal contact lens to dry eye degree of flight attendants.
METHODS:Eighty cases (160 eyes) of flight attendants from China Southern were divided into two groups:control group 40 cases ( 80 eyes ) wearing ready-made Bausch soft corneal contact lens ( curvature 8. 4 ); the experiment group 40 cases ( 80 eyes ) , wearing Bausch soft corneal contact lens with customized curvature. Tear break-up time ( BUT ) , Schirmer Ⅰ test ( SⅠt ) and fluorescein ( FL ) staining were as dry eye evaluation index. The results was statistically analyzed.
RESULTS: BUT, SⅠt average shortening value of the experimental group were less than that of the control group, there was statistical significance (P<0. 01). FL staining positive increase, the number of experimental group was fewer than that of control group, with statistical significance (P<0. 05).
CONCLUSION: Wearing customized curvature soft corneal contact lens can prevent the flight dry eye more effectively than fixed curvature product.

Objective To detect antibodies against chlamydial plasmid-encoded protein 3 (Pgp3),outer membrane complex protein B C-terminal peptide (OmcBc),CT841 protein and heat shock protein 60 (HSP60) in the sera of patients with urogenital Chlamydia trachomatis infection.Methods Recombinant plasmids encoding the aforementioned four proteins and an empty plasmid were transformed into Escherichia coli separately followed by 2-hour isopropyl-1-thio-β-galactopyranoside (IPTG) induction and cell lysis.The expressed proteins were purified with glutathione magnetic beads and then used to coat 96-well enzyme-linked immunosorbent assay (ELISA) plates precoated by glutathione.Serum samples were collected from 20 patients with and 20 clients without urogenital C.trachomatis infection attending the sexually transmitted disease (STD) clinic of Tianjin Medical University General Hospital.ELISA with the expressed protein-coated plates was adopted to detect antibodies against these proteins in the serum samples.Results Of the 20 serum samples from C.trachomatis-infected patients,14 (70％)had anti-Pgp3 antibody,9 (45％) anti-OmcBc antibody,8 (40％) anti-CT841 antibody,and 5 (25％) anti-HSP60 antibody.Meanwhile,no antibody was detected in any of the serum samples from uninfected clients except for one with anti-HSP60 antibody.Conclusions Of the four studied C.trachomatis proteins,Pgp3 appears to have the strongest antigenicity with the highest antibody-detection rate,while HSP60 exhibits the weakest antigenicity with the lowest antibody-detection rate.

Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.

Objective To evaluate the clinical significance of high frequency ultrasound in the diagnosis of duodenal obstruction in neonates.Methods Ultrasonography,clinical data and etiological diagnoses of the operation in 113 neonates with duodenal obstruction were reviewed retrospectively.The digestive tract,including stomach,duodenum,jejunoileum and colon,were examined in all patients with 8-12 MHz linear transducer before operation.Results In the 113 neonates with duodenal obstruction,63 cases were diagnosed intestinal malrotation,31 cases duodenal stenosis,14 cases annular pancreas,and 5 cases duodenal atresia.One hundred and six cases were diagnosed as duodenal obstruction by ultrasound,of which intestinal malrotation in 61 cases,duodenal stenosis in 29 cases,duodenal atresia in 4 cases,and annular pancreas in 12 cases.The diagnostic rate was 93.81% (106/113 cases),96.83% (61/63 cases),93.55% (29/31 cases),80.00% (4/5cases) and 85.71% (12/14 cases),respectively.The location of obstruction diagnosed by ultrasound was coincident with the operation in 92 cases,with a diagnostic rate of 81.42%(92/113).Conclusions High frequency ultrasound plays an important role in diagnosing the causes and location of duodenal obstruction.It can be used as the first choice of examinal methods for the neonates with duodenal obstruction.

AIM: To investigate the feasibility and effect of directly differentiation of embryonic stem cells(ESC) into neural cells induced by retinoid acid(RA) without embryonic body(EB) culture period in vitro.METHODS: ESC were digested and divided into 4 groups: group A and B were undergone EB culturing.After that,cells in group A were induced by RA,cells in group B were differentiated spontaneously,cells in group C were committedly induced by RA directly without EB culturing,and cells in group D were differentiated spontaneously without EB period.Morphologic changes were observed under inverted microscope and scanning electron microscope.MAP-2 and GFAP were detected by immunocytochemistry and flow cytometry after differentiated for 9 days.RESULTS: In groups A or C,neuron-like cells increased gradually,forming neural network.At the 9th day,a large part of cells in these groups were MAP-2 positive cells,and the positive rate was higher than that in groups B or D(P0.05).CONCLUSION: ESC was directly induced into neural cells by RA without EB culture period in vitro.This modified method has the same effect as the traditional RA 4-/4+ assay and can replace the traditional method.

OBJECTIVETo explore the effect of ERK1/2 MAPK pathway on the expression of Kv1.5 channel, a voltage-gated potassium ion channel, in rat pulmonary artery smooth muscle cells (PASMCs) and its mechanisms during the process of hypoxia.

METHODSThe PASMCs derived from SD rats were cultivated primarily. The third to sixth generation of PASMCs were divided into 5 groups randomly: (1) Normal group (N); (2) Hypoxic group (H); (3) Demethy sulfoxide(DMSO) group (HD); (4) U0126 group (HU): 10 micromol/L U0126; (5) Anisomycin group (HA): 10 micromol/L anisomycin. There were three dishes of cells in each group. The cells in normal group were cultured in normoxic incubator (5% CO2, 37 degrees C), the cells in other groups were added to 0.05% DMSO in the hypoxic incubator (5% CO2, 2% O2, 37 degrees C), all cells were cultured for 60 h. RT-PCR and Western blot were used to detected the espressions of Kv1.5 mRNA and protein in PASMCs.

RESULTSCompared with N group, the expressions of Kv1.5 mRNA and protein in H, HD and HA groups were reduced significantly (P < 0.05); Compared with H group and HD groups, Kv1.5 mRNA and protein expressions in HU group were increased sharply (P < 0.05). Compared with the HU group, Kv1.5 mRNA and protein expressions in HA groups were significantly lower (P < 0.05).

CONCLUSIONLow oxygen reduced Kv1.5 mRNA and protein expressions, U0126 could resistant the Kv1.5 channel lower expression caused by hypoxia. Anisomycin had no significant effect on Kv1.5 channel expression under hypoxia, but the expression of Kv1.5 was still significantly lower than the normal oxygen group. These data suggest that hypoxia may cause hypoxic pulmonary hypertension by interfering ERK1/2 signaling pathway to inhibit Kv1.5

Animals ; Cell Hypoxia ; Hypertension, Pulmonary ; Kv1.5 Potassium Channel ; metabolism ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; Oxygen ; Pulmonary Artery ; cytology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley