Objective To construct a recombinant plasmid encoding Tp0821,a membrane lipoprotein of T. pallidum,express and purify this protein,and to evaluate its immunocompetence.Methods The recombinant plasmid pQE32/Tp0821 was constructed and induced to express the corresponding protein.Then,New Zealand rabbits were immunized with purified recombinant protein to prepare polycional antibodies,and the titer of polyclonal antibody was determinated.Indirect ELISA was developed with the recombinant protein of T. pallidum as coating antigen to detect 80 control sera and 150 FTA-ABS-positive sera.Results The recombinant plasmid pQE32/Tp0821 was constructed and a fusion protein with expected molecular weight was expressed.Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the antibody titer reached 1:6400.Compared with FTA-ABS test,the indirect ELISA showed a sensitivity and specificity of 92.6%and 98.6%,respectively,in the detection of control and clinical sera.Conclusion The recombinant protein Tp0821 shows excellent immunocompetence,which can be applied to the serological diagnosis of syphilis.

Objective To clone, express Tp0319 gene from Treponemapallidum (T. pallidum), and to assess the immunocompetence of recombinant protein. Methods The immuno-dominant region of Tp0319gene was chosen by computer analysis, amplified from T. pallidum complete genome by PCR, subcloned into the expression vector pQE32 to construct a recombinant plasmid, pQE32/Tp0319, which was then expressed in E. coli M15. The recombinant protein was purified with Ni-NTA affinity chromatography, and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot. New Zealand rabbits were immunized with the recombinant protein, and the titer of anti-Tp0319 antibodies in sera from immunized rabbits were measured with indirect ELISA. Also, indirect ELISA with the recombinant Tp0319 as coating antigen was performed to detect the anti-Tp0319 antibody in sera from 200 normal human controls and 200 patients with syphilis. Results The prokaryotic expression vector pQE32/Tp0319 was constructed successfully, and the recombinant protein Tp0319 with a molecular weight of about 30 000 was attained. Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the specific antibody titer was more than 1: 10 240 after immunization for 3 times. Western blot proved that the recombinant protein could specifically react with anti-T. pallidum IgG antibody-positive sera. Indirect ELISA was successfully developed with the recombinant Tp0319, and detected antibodies to T. pallidum in control sera with a sensitivity and specificity of 100% (40/40), respectively. Compared with T. pallidum particle agglutination (TPPA) assay, the sensitivity and specificity of the indirect ELISA were 92.6% and 100%, respectively, in the detection of T. pallidum in sera from patients and controls, and the concordance between the indirect ELISA and TPPA was 96%. Conclusions The prepared recombinant protein shows a satisfactory immunocompetence, which may lay a foundation for its further application in the serodiagnosis of syphilis.

Objective To investigate the immune response to and protective effect of a bivalent DNA vaccine expressing interleukin-2(IL-2)and Gpd proteins in New Zealand rabbits.Methods Seventy-two male New Zealand white rabbits were equally and randomly divided into 4 groups to be immunized with recombinant plasmids pcDNA3.1(+)/Gpd-IL-2(pcD/Gpd-IL-2),pcDNA3.1(+)/Gpd(pcD/Gpd),empty plasmid pcDNA3.1(+)(pcD)and phosphate buffered saline(PBS),respectively.Immunization was carried out by intramuscular injection at multiple sites with a 2-week interval for 3 times.On week 10 after the initial immunization,the rabbits were challenged intradermally with T.pallidum(Nichols strain).Enzyme-linked immunosorbent assay(ELISA)was used to quantify the serum level of anti-Gpd antibodies in the rabbits and the level of IL-2 and interferon(IFN-γ)in the supernatant of Gpd protein-stimulated spleen cells from the rabbits at different time pionts.MTT assay was conducted to detect the proliferation response of spleen cells collected from the rabbits on day 0,14,28,140 and 168 after the challenge.Results Compared with pcD and PBS,both the vaccines pcD/Gpd and pcD/Gpd-IL-2 elicited significantly higher levels of anti-Gpd IgG antibodies in rabbits at different time points during the vaccination and infection period,with the titers peaking at 1 ∶ 1024 and 1∶4096,respectively(both P ＜ 0.01).There were also significant differences in the serum levels of anti-Gpd IgG antibodies between the pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits at different time points(all P ＜0.01).The levels of IL-2 in the supematant of spleen cells from pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits on week 8 after the immunization were 110 ± 12.6 and 167 ± 15.7 μg/L respectively,and those of IFN-γwere 225 ± 17.6 and 447 ± 22.4 μg/L respectively,significantly higher than those in that from the other two groups of rabbits(all P ＜ 0.01).Furthermore,an apparent proliferation response was observed in spleen cells from pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits with a higher stimulation index compared with pcD-and PBS-immunized rabbits(all P ＜ 0.01).Dark-field microscopic examination of early-stage infected lesions revealed that pcD/Gpd-IL-2-immunized rabbits had a lower detection rate(17.5％)of Tp from lesions,occurrence of ulcerative lesions(15％)and shorter curing time compared with pcD/Gpd-immunized rabbits.Conclusion The recombinant plasmid pcDNA3.1(+)/Gpd-IL-2 could induce protective humoral and cellular immune response more efficiently in rabbits.

Objective The mechanism of traditional double-eyelid blepharoplasty is adhering skin to tarsus,by which the skin below the incision could be elevated along with the tarsus and produce the crease.But the anatomic basis of native double-eyelids is not the same,which depends on the connection of levator aponeuroses with muscle and skin.That is,the elevations of tarsus and skin is based on the different branches of levator aponeuroses to skin.According to this,the authors have developed a modified blepharoplasty suturing levator aponeuroses to the obicularis occuli muscle to mimic the native double eyelid.Methods The surgical procedures were modified from traditional surgery.The major improvements were:keeping obicularis occuli muscles intact,dissecting the levator aponeuroses and suturing them together.Since the suturing was performed between mobilized tissues of muscle and aponeurosis,the method was named as "flexible suspension technique".Five-hundred patients were performed with this method.Forty cases randomly selected from them were compared with 40 cases performed with traditional method,and the results were investigated using questionnaire survey.Results The results of the modified surgery were more natural than traditional technique.Majority of patients were satisfactory,whereas 15 cases (3％) had asymmetry,narrowing and even fold loss.All these 15 cases were puffy eyelids.Statistical results indicated that the satisfaction rate was significantly high (P =0.003),and the adverse effects (edema,bruise and scar) were significantly lighter than traditional technique;whereas the asymmetry and fold loss were higher than traditional technique.Conclusions The flexible suspension technique of blepharoplasty can obtain more natural double eyelid appearance,and is suitable for primary blepharoplasty but not applicable for severe puffy eyelids.

Objective To investigate the efficiency and safety of allogeneic hematopoietic cell transplantation for malignant hematological diseases in patients older than 50 yeas of age. Methods From May 2002 to January 2010, 35 patients (＞ 50 years) with malignant hematological diseases received allogeneic hematopoietic cell transplantation. In 35 patients, 18 patients were conditioned with non-myeloablative regimen and 17 patients with myeloablative regimen. The outcome,engraftment and prognosis of allogeneic hematopoietic cell transplantation were analyzed. Results The hematopoetic reconstitution was achieved in 32 of 35 patients. The median time of granulocyte count exceeding 0. 5 × 109/L was 12 days and the that of platelet count exceeding 20 × 109/L was 17days. The cumulative incidence of aGVHD was 48. 6 %, and 37. 9 % patients developed cGVHD.The estimate probability of cumulative survival at 5 years was 48. 5 %, The estimate probability of cumulative mortality rate was 51.5 %, and the estimated transplant-related mortality was 22. 9 %.The relapse rate was 11.4 %. There was significant difference except for the incidence of cGVHD.Conclusion Allogeneic hematopoietic cell transplantation may be appropriate for older patients with malignant hematological diseases.

Objective:To figure out the structure and relevant data measurements of zygomatic ligament by cadaver anatomy and review of previous studies.Methods:From July 2018 to January 2020, the zygomatic areas of 20 Chinese frozen fresh cadaver hemifaces were dissected in the Department of Anatomy, Health Science Center of Hangzhou Normal University. Then the structures of zygomatic ligaments were shown. The characters of the ligament and the relationship with adjacent tissue were described and measured. And 16 previous studies were reviewed to get a comprehensive description about the characters of zygomatic ligaments.Results:Zygomatic ligaments were even and dense fibrous tissue structures distributed vertically between the skin and the subcutaneous tissue. Under the SMAS plane, the ligaments divided into two bundles. The origin of major bundle located beyond the origin of the zygomatic major muscle on the periosteum, and the origin of minor bundle located between the origin of the zygomatic minor and major muscle.Conclusions:The anatomy of the zygomatic ligament has a regular pattern, and its anatomical data has certain directive significance for clinical application.

Objective:To identify the association of sex hormone and sex hormone-binding globulin (SHBG) levels in patients with Graves′ disease.Methods:Between December 2017 and July 2019, 152 patients with Graves′ disease were enrolled in the Department of Endocrinology and Metabolism of Zhongshan Hospital, Fudan University. Parameters such as height, weight, thyroid function, sex hormone, fasting blood glucose, insulin, C-peptide, glycosylated albumin, HbA 1C, and liver function were collected. The associations between SHBG and bioclinical characteristics were analyzed. Results:The glycosylated albumin level was negatively associated with SHBG in all subjects ( β=-0.308, P<0.01), while this association was not significant after the adjustment for thyroid hormones. Male patients had significantly lower SHBG level than female patients ( P<0.01). In male patients, SHBG was associated with luteinizing hormone ( r=0.465, P<0.01), estradiol ( r=0.629, P<0.01), testosterone ( r=0.786, P<0.01). While in female patients, SHBG was also associated with testosterone ( r=0.191, P<0.05). In multivariate linear regression analysis, estradiol was independently associated with SHBG ( β=0.329, P<0.01) in male subjects. Conclusion:The hypogonadism and infertility in Graves′ disease patients could partially be attributed to the association between SHBG and Graves′ disease.

Hyaluronic acid (HA) injections have already been one of most popular cosmetic procedures for around 18 years. However, blindness is one of the most serious complications caused by HA injections. There is still no consensus on the treatment for vision recovery. The efficacy and safety of different delivery administration of hyaluronidase such as retrobulbar injection and endovascular intervention are still controversial so far. Based on current status, prevention is prior to treatment, which is widely recognized in the world. The pathogenic mechanism of blindness induced by filler injection is still not clear, and there exist several theories including the artery embolism, ischaemia, artery spasm and venous involvement, which are all related to the ophthalmic artery and its branches. Therefore, acquiring anatomic knowledge and the relevant parameters will help us prevent to damage the ophthalmic artery and its branches during HA injections. The mechanism, treatment progress and prognosis of blindness caused by HA injections in recent years were analyzed and summarized in this article. We hope it will help the clinician to improve the relevant knowledge about vision loss caused by HA injection, as well as to promote the safety of filler injection.

Hyaluronic acid (HA) injections have already been one of most popular cosmetic procedures for around 18 years. However, blindness is one of the most serious complications caused by HA injections. There is still no consensus on the treatment for vision recovery. The efficacy and safety of different delivery administration of hyaluronidase such as retrobulbar injection and endovascular intervention are still controversial so far. Based on current status, prevention is prior to treatment, which is widely recognized in the world. The pathogenic mechanism of blindness induced by filler injection is still not clear, and there exist several theories including the artery embolism, ischaemia, artery spasm and venous involvement, which are all related to the ophthalmic artery and its branches. Therefore, acquiring anatomic knowledge and the relevant parameters will help us prevent to damage the ophthalmic artery and its branches during HA injections. The mechanism, treatment progress and prognosis of blindness caused by HA injections in recent years were analyzed and summarized in this article. We hope it will help the clinician to improve the relevant knowledge about vision loss caused by HA injection, as well as to promote the safety of filler injection.

Objective To investigate imaging manifestations of desmoids-type fibromatoses (DF).Methods The CT and MRI features of 17 cases with DF were reviewed retrospectively.Resuits All of 17 cases were solitary lesions,of which 10 cases were abdominal wall shape,5 cases were abdominal wall type and 2 cases were intra-abdominal (mesentery) type.In all the cases,10 times CT and 10 MRI examinations were performed,of which 3 cases underwent CT and MRI examination.In 10 cases with CTexamination,9 cases of plain scan was density,uniform density,1 cases of abdominal DF scan showed slightly higher local density.Enhanced CT scanning was performed in 5 cases,including 4 cases of mild enhancement,and the other 1 case of intra-abdominal DF showed annular enhancement.2 cases of delayed scan showed progressive enhancement.MRI examination was performed in 10 cases,T1WI were equal signals,9 cases of T2WI and SPIR sequences were high signals,and internal signals were not uniform.One case of internal DF showed patchy,short T2 low signal,and SPIR was low signal.6 cases of DF underwent enhanced scan,the lesions showed moderate and markedly inhomogeneous enhancement,with punctate,funicular and patchy enhancement in the interior zone.Conclusion The CT and MRI features of DF have some characteristics,and MRI can reflect the histological features of lesions.