The recombinaut porcine insulin precursor(PIP)produced by Pichia pastoris in shake-flask and 501.fermenter was investigated respectively.The results indicated that 60h induction time length and 2.0%～2.5% methanol addition every day was optimum in shake- flask.The process in 50L fermenter was consisted of batch,feed-batch and induction phases.The relationship between dry cell weight(y) and culture time (t) in growth phase(batch and feed-batch phase)could be described by model y=0.6525e~(0.1907t).Glycerol and ammonia were almost used for cell growth and maintain,and no by-product was observed in batch and fed-batch phase Only 80% ammonia and 70% methanol were used by cell in induction phase.By comparison the results of shake-flask and 50L fermenter,it was concluded that the limit- ing factor in the fermentation of shake-flask and 50L fermenter was dissolved oxygen(DO)and.carbon source,respectively.When scaling the result of shake-flask to 501.fermenter,the control strategy was adapted for 50L fermenter by increasing the feed rate of methanol and the maximum PIP concentration reached 1.72 g/L.

OBJECTIVETo investigate possible mechanism of angiogenesis in brain tissue of neonatal rat hypoxic-ischemic encephalopathy (HIE).

METHODSForty seven-day old neonatal rats were randomly assigned to hypoxic-ischemic (Model group) or sham treatment (Sham group), each group had 20 rats. Five rats from each group were sacrificed on days 1, 3, 7 and 14 after hypoxia-ischemia. Paraffin sections of the brain were stained with anti-endothelial cell, anti-proliferating cell nuclear antigen (PCNA) or anti-vascular endothelial growth factor (VEGF) by using single or double immunohistochemistry. The brain capillary density index (BCDI), brain proliferating capillary density index (BPCDI) and the expression of VEGF were analyzed under the microscope. The expression of VEGF and hypoxia-inducible factor-1alpha (HIF-1alpha) mRNA in hypoxic-ischemic side of the brain was measured by RT-PCR.

RESULTSBCDI around infarct brain tissue in the model group began to rise on day 3 and remained higher than that of the sham group from day 3 to day 14 [day 3: (9.80 +/- 1.05)/HPF vs. (4.90 +/- 0.66)/HPF, P < 0.01;day 14: (13.29 +/- 3.90)/HPF vs. (6.08 +/- 1.50)/HPF, P < 0.01]. Occasional proliferating capillary was found in brain tissue of normal neonatal rats. The density of proliferating brain capillary on day 3 and day 7 of Model group [(0.54 +/- 0.15)/HPF vs. (0.90 +/- 0.25)/HPF] were significantly higher than those of Sham group [(0.12 +/- 0.05)/HPF vs. (0.13 +/- 0.07)/HPF, P < 0.01]. VEGF was mainly expressed in the cytoplasm of neurons, capillary endothelial cells and pial cells. Viable neurons and endothelial cells in the infarct areas also expressed VEGF. The expression of VEGF mRNA in hypoxic-ischemic brain tissue was significantly higher than that of normal control (P < 0.01) and temporally preceded angiogenesis. The expression of VEGF mRNA at 12 hours of HIE model was significantly higher than that of normal control (1.56 +/- 0.27 vs. 0.95 +/- 0.21, P < 0.05). It reached its peak on day 1 and day 3 (1.85 +/- 0.31 vs. 1.86 +/- 0.39), significantly higher than that of normal control (P < 0.01), and decreased by day 7 and day 14, without significant difference compared with normal control (P > 0.05). The expression of HIF-1alpha mRNA was also up-regulated after hypoxic-ischemic treatment. The expression of HIF-1alpha mRNA (1.07 +/- 0.21) was significantly higher than that of normal control (0.64 +/- 0.28, P = 0.048) at 3-hour of HIE model, reached its peak on day 1 (1.73 +/- 0.42, P < 0.01), remained at high expression level on day 3 (1.44 +/- 0.36, P < 0.05) and began to decline by day 7 and day 14 when it was not significantly different from normal control.

CONCLUSIONSAngiogenesis exists in the brain tissue of neonatal rat HIE model. Up-regulation of VEGF expression mediated by HIF-1 may play an important role in the process of angiogenesis.

Animals ; Animals, Newborn ; Brain ; blood supply ; Brain Diseases ; etiology ; genetics ; metabolism ; Disease Models, Animal ; Hypoxia-Inducible Factor 1, alpha Subunit ; Hypoxia-Ischemia, Brain ; complications ; Immunohistochemistry ; Neovascularization, Pathologic ; etiology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; analysis ; genetics ; Vascular Endothelial Growth Factor A ; analysis ; genetics

On the base of element and metablism balancing, the mathematical model of the cultivation process with Streptomyces aureofaciens was developed, and the unknown parameters in the model were estimated with the method of nonlinear optimization. Firstly the energetic coefficient of CTC biosynthesis was gained, which was 1.8 - 2.8 mol-ATP x C-mol(-1). The macroscopic reaction rates were predicted in the process and compared with the experimental values. The results show that the model can preferably describe the relationships between several macroscopic reaction rates in the process and can supervise the optimization of CTC fermentation process theoretically.
Chlortetracycline ; metabolism ; Fermentation ; physiology ; Models, Theoretical ; Streptomyces aureofaciens ; growth & development ; metabolism

OBJECTIVE: To investigate the tumor regression and local immune function in nasopharyngeal carcinoma patients treated with p53 gene therapy. METHOD: The two-step immunohistochemical was done to detect the expression of tumor-infiltrating lymphocytes (TIL) T-cell receptor-CD3, CD4, CD8 and B cell receptor-CD20 in the primary tumor tissue of nasopharyngeal carcinoma. Nasal endoscopy with MRI or CT was used for evaluation of tumor size. RESULT: The expression of CD3, CD4, CD8 was significantly increased after p53 gene treatment (P < 0.05). There was no significant change in expression of CD20 after p53 gene treatment (P > 0.05). In conventional treatment group, CD3, CD4, CD8 and CD20 (P > 0.05) did not show any significant difference. In gene therapy group at 3 months after treatment, 20 patients had achieved CR, 10 PR, 1 SD, 1 PD. In conventional treatment group, 11 patients had achieved CR, 12 PR,5 SD,3 PD. The response rate between treatment group and control group (CR+PR) was different (P < 0.05). CD3 and CD4 expression was correlated with tumor regression rate (P < 0.05, P < 0.01), and CD8 expression was correlated with the CR rate (P < 0.05). CONCLUSION T cells are the most proliferative cell of TII. in NPC patients after p53 gene therapy The local cellular immune status is positively correlated with tumor regression rate.
Adult ; Aged ; CD4-Positive T-Lymphocytes ; immunology ; Carcinoma ; Female ; Genes, p53 ; Genetic Therapy ; Humans ; Lymphocyte Count ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; immunology ; pathology ; therapy

OBJECTIVE To study the clinic feature and cause of misdiagnosis of early stage syphilis and evaluate the significance of histopathology in the diagnosis of the disease. METHODS Totally 1 200 early syphilis cases were analyzed.The serologic test for syphilis was performed.Thirty five of them were performed with histopathological examination. RESULTS The primary syphilis was found to be commonly misdiagnosed as chancroid,genital herpes,scabies nodules and ulcus vulvae acutum.For secondary syphilis,macular syphilide and maculopapular syphilide were easily misdiagnosed as pityriasis rosea or dermatitis.The papulosquamous syphilide was commonly misdiagnosed as psoriasis.The condyloma latum was commonly misdiagnosed as condyloma acuminatum. CONCLUSIONS The serologic test is important in diagnosis of primary syphilis.The histopathologic test plays a role in diagnosis of primary syphilis,condyloma latum and papulosquamous syphilide,but of limited value in diagnosis of macular syphilide.

To evaluate the papilla alterations around single-implant restorations in the anterior maxillae after crown attachment and to study the influence of soft tissue thickness on the papilla fill alteration. According to the inclusion criteria, 32 patients subjected to implant-supported single-tooth restorations in anterior maxillae were included. The patients were assigned to two groups according to the mucosal thickness: (i) group 1, 1.5 mm s mucosal thickness 3 mm; and (ii) group 2, 3 mm Adult ; Crowns ; Dental Implantation, Endosseous ; methods ; Dental Implants, Single-Tooth ; Dental Prosthesis Design ; Dental Prosthesis, Implant-Supported ; Esthetics, Dental ; Female ; Follow-Up Studies ; Gingiva ; anatomy & histology ; Humans ; Male ; Maxilla ; Middle Aged ; Outcome Assessment (Health Care) ; Prospective Studies ; Tooth Cervix ; anatomy & histology ; Young Adult

BACKGROUND: It remains unclear whether the outcomes of ST-elevation myocardial infarction (STEMI) patients treated with primary percutaneous coronary intervention (PPCI) during off-hours are as favorable as those treated during on-hours, especially those with a first medical contact-to-device (FMC-to-device) time within 90 min. We aimed to determine whether off-hours admission impacted late outcomes in patients undergoing PPCI and with an FMC-to-device time ≤90 min. METHODS: This multicenter retrospective study included 670 STEMI patients who underwent successful PPCI and had an FMC-to-device time ≤90 min from 19 chest pain centers in Beijing from January 2018 to December 2018. Patients were divided into on-hours group and off-hours group based on their arrival time. Baseline characteristics, clinical data, and key time intervals during treatment were collected from the Quality Control & Improvement Center of Cardiovascular Intervention of Beijing by the "Heart and Brain Green Channel" app. RESULTS: Overall, the median age of the patients was 58.8 years and 19.9% (133/670) were female. Of these, 296 (44.2%) patients underwent PPCI during on-hours and 374 (55.8%) patients underwent PPCI during off-hours. Compared with the on-hours group, the off-hours group had a longer FMC-to-device time and fewer patients with FMC-to-device time ≤60 min (P < 0.05). During the mean follow-up period of 24 months, a total of 64 (9.6%) participants experienced a major adverse cardiovascular event (MACE), with 28 (9.1%) in the on-hours group and 36 (9.6%) in the off-hours group (P > 0.05). According to the Cox regression analyses, off-hours admission was not a predictor of 2-year MACEs (P = 0.788). Similarly, the Kaplan-Meier curves showed that the risks of a MACE, all-cause death, reinfarction, and target vessel revascularization were not significantly different between the two groups (P > 0.05). CONCLUSIONS This real-world, multicenter retrospective study demonstrated that for STEMI patients who underwent PPCI within 90 min, off-hours admission was safe, with no difference in the risk of 2-year MACEs compared with those with on-hours admission.
Beijing ; Female ; Humans ; Middle Aged ; Percutaneous Coronary Intervention ; Retrospective Studies ; Risk Factors ; ST Elevation Myocardial Infarction/surgery* ; Treatment Outcome

To evaluate the papilla alterations around single-implant restorations in the anterior maxillae after crown attachment and to study the influence of soft tissue thickness on the papilla fill alteration. According to the inclusion criteria, 32 patients subjected to implant-supported single-tooth restorations in anterior maxillae were included. The patients were assigned to two groups according to the mucosal thickness: (i) group 1, 1.5 mmfmucosal thicknessf3 mm; and (ii) group 2, 3 mm,mucosal thicknessf4.5 mm. Assessments of interproximal papillae at the time of crown placement (baseline) and at 6-month postloading (follow-up) were made by two prosthodontists using papilla fill index (PFI). The mean mucosal thickness was (2.4960.31) mm(group 1) and (3.8160.31)mm (group 2) for the two groups respectively. A significant difference in PFI between the groups was detected at the baseline (P,0.001). PFI improvements over time occurred after 6-month follow-up irrespective of the groups. When compared to group 1, the likelihood to obtain papilla fill was significantly higher for group 2 with an odds ratio of 6.05 (P,0.001). The interproximal papilla level around single-implant restorations could improve significantly over time after 6-month restoration according to PFI assessment. The thicker mucosa before implant placement implied a more favorable esthetic outcome in papilla alteration.

[Objective]To discuss the CT diagnosis and differential diagnosis of solid pseudopapillary tumor of the pancreas (SPTP).[Methods]The CT findings of 20 patients with SPTP proved by surgically pathology were retrospectively analyzed and summa?rized.[Results]SPTP were composed of solid and cystic components with surrounding capsule resulting to clear demarcation between tumor and normal pancreas without dilation of pancreatic duct. The tumor parenchyma was slightly hyperenhancement on arterial phase and showed gradual enhancement on venous and delayed phase.[Conclusions]The CT findings of SPTP have relative specifici?ty and can contribute to early diagnosis and differential diagnosis of SPTP.

OBJECTIVE: To investigate a family with congenital dysfibrinogenemia, and analyze the risk of hemorrhage and thrombosis and blood transfusion strategies. METHODS: Prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT) of the proband and her family members were detected by automatic coagulometer, fibrinogen (Fg) activity and antigen were detected by Clauss method and PT algorithm respectively. Meanwhile, thromboelastometry was analyzed for proband and her family members. Then, peripheral blood samples of the proband and her family members were collected, and all exons of FGA, FGB and FGG and their flanks were amplified by PCR and sequenced to search for gene mutations. RESULTS: The proband had normal APTT and PT, slightly prolonged TT, reduced level of Fg activity (Clauss method). The Fg of the proband's aunt, son and daughter all decreased to varying degrees. The results of thromboelastogram indicated that Fg function of the proband and her family members (except her son) was basically normal. Gene analysis showed that there were 6233 G/A (p.AαArg35His) heterozygous mutations in exon 2 of FGA gene in the proband, her children and aunt. In addition, 2 polymorphic loci were found in the family, they were FGA gene g.9308A/G (p.AαThr331Ala) and FGB gene g.12628G/A (p.BβArg478Iys) polymorphism, respectively. The proband was injected with 10 units of cryoprecipitate 2 hours before delivery to prevent bleeding, and no obvious bleeding occurred during and after delivery. CONCLUSION Heterozygous mutation of 6233G/A (p.AαArg35His) of FGA gene is the biogenetic basis of the disease in this family with congenital dysfibrinogenemia.
Humans ; Child ; Female ; Fibrinogen/genetics* ; Pedigree ; Afibrinogenemia/genetics* ; Mutation ; Blood Transfusion