Objective To trace changes in the transcript level of the Treponema pallidum(Tp)protein Tp0751 in skin lesions of a rabbit model of early syphilis. Methods Three New Zealand white rabbits were intracutaneously injected with 0.1 ml of Tp (Nichols Seattle strains)suspensions (107 treponemes/ml)at 10 sites on the shaved back to establish a model of early syphilis. All the rabbits received a single injection with the total amount of treponemes being 107. Then, skin changes at injection sites were observed, and the size of skin rashes was recorded on a daily basis. Skin specimens sized 0.4 cm × 0.4 cm were excised from an injection site and a non-injection site(negative control)separately every 3 days for the detection of Tp0751 and Tp0574 mRNAs. The whole experiment lasted 30 days, and a total of 11 skin biopsies were carried out. Fluorescence-based quantitative PCR was performed to measure the mRNA expressions of Tp0751 and Tp0574 continuously and dynamically during the development of chancre. Results After intracutaneous injection of Tp suspensions, red papules occurred on the back of rabbits on day 6, and reached maximum size on day 19 with the formation of ulcer and chancre. On day 25, disseminated secondary syphilides gradually appeared all over the body surface of the rabbits. The mRNA expression levels of Tp0574 and Tp0751 increased at the early stage, peaked onday 15 (compared with the other time points, all P < 0.05), thereafter rapidly declined, but rose slightly on day 27. The standardized expression level of Tp0751 mRNA increased gradually after day 15, and peaked on day 24 (compared with the other time points, all P < 0.05). Conclusion The transcript level of Tp0751 was high in rabbits at the late stage of Tp clearance when generalized disseminated secondary syphilides had not appeared, suggesting that Tp0751 may be involved in the systemic spread of Tp.

Objective To explore the effects of improvements on setup error ( SE ) and clinical target ( CTV ) margin of supraclavicular field ( SCF ) by using moisture-cured resin cushion and breast bracket for lower neck fixation in breast cancer patients who underwent post-mastectomy radiotherapy. Methods Totally 13 patients with breast cancer who underwent post-mastectomy radiotherapy were enrolled. All patients were immobilized by breast bracket and moisture-cured resin cushion. Firstly, each patient′s lower neck and head was fixed well by moisture-cured resin cushion, filling the gap between the neck and breast bracket. Secondly,each patient underwent the cone-beam CT ( CBCT) at the first, tenth and twentieth treatment after positioning. Then these CBCT images were registered to the planning CT to determine setup errors in translational and rotational directions, and SCF CTV margins by the systematic and random errors were evaluated. Results The setup errors in x (left-right), y (superior-inferior), z (anterior-posterior) translational directions were (2. 16 ± 1. 25), (1. 50 ± 1. 28), (1. 94 ± 1. 12) mm and (1. 76 ±1. 87)°, (1. 82 ±1. 12)°, (0. 99 ±0. 58)°, respectively in θ (pitch degree),Ф(roll degree),ψ( yaw degree) rotational directions. Non-parametric rank test ( Mann-Whitney U test) was performed with previous data, the differences of the setup error in y, z,θ directions were statistically significant ( Z =4. 152, 3. 415, 2. 053, P<0. 05). The margins from CTV were 4. 07, 4. 03 and 3. 73 mm in x, y and z directions, respectively. Compared with the previous study on SCF, CTV margin required 8, 8 and 6 mm in x, y and z axis directions, the volume of CTV to PTV were decreased by 32. 73% on average. Conclusions Compared with plastic circular pillow alone, moisture-cured resin cushion with breast bracket locating method could reduce setup errors in SCF target region. The margins from CTV to PTV weredecreased to 4. 07, 4. 03 and 3. 73 mm in x, y and z axis directions at least.

Objective To isolate and culture a clinical strain (GDI) of Treponema pallidum (Tp) in Guangdong province,and to investigate the difference in nonsynonymous single nucleotide polymorphisms (nsSNPs) between the GD1 strain and Tp Nichols strain.Methods The GD1 strain was isolated from the hard chancre in a patient with primary syphilis in Guangdong province,and continuously subcultured in the testes of New Zealand white rabbit.The serial subcultivation of GD1 was multi-verified by dark-field microscopy,polymerase chain reaction (PCR) for TP0548 gene,DNA sequencing and genotyping.Meglumine diatrizoate density gradient centrifugation was performed to isolate rabbit tissues and concentrate GD1,and DNA sequencing was used to verify the nsSNPs in the TP0443 and TP0584 genes.Results The GD1 strain was successfully isolated from the lesions of the patient with syphilis,and classified as a subtype f of TP0548.Compared with the American Tp (Nichols strain),there were nsSNP mutations in the GD1 strain.One mutation was located in the TP0443 gene,leading to the the substitution of threonine by alanine at amino acid position 120,and another one was located in the TP0584 gene,which caused a change from alanine to threonine at amino acid position 314.Conclusion The GD1 strain was successfully isolated firstly from the lesions in a patient with syphilis in Guangdong province,and nsSNP mutations were confirmed in the GD 1 strain on the etiology.

Objective:To evaluate the relationship of early tumor shrinkage and depth of tumor response with the clinical efficacy and pro-gnosis of programmed death-1(PD-1)inhibitor combined with trastuzumab and first-line chemotherapy in the treatment of HER-2-positive advanced gastric cancer.Methods:We retrospectively analyzed data from 40 patients treated with this combination at Nanjing Drum Tower Hospital,The Affliated Hospital of Nanjing University Medical School from June 2018 to March 2023.Key metrics included early tumor shrinkage(ETS),depth of response(DpR),objective response rate(ORR),disease control rate(DCR),progression-free survival(PFS),overall survival(OS),and adverse reactions.Survival analysis using Log-rank test and Kaplan-Meier method,and plot PFS and OS survival curves.COX regression analysis for correlation testing.Results:The patient's ORR was 77.5%,DCR was 100%,complete response rate was 15.0%,medi-an PFS(mPFS)was 11.10 months,and median OS(mOS)was 30.77 months.COX univariate analysis showed that tumor differentiation,liver metastasis,distant lymph node metastasis,DpR were related to PFS and OS(all P<0.05),ETS was only related to PFS(P=0.010),and PD-L1 ex-pression was not related to PFS and OS.There was a significant difference in mPFS between patients with ETS≥35%and ETS<35%(P=0.008),while there was no significant difference in mOS(P=0.076);There were significant differences in mPFS and mOS between patients with DpR≥40%and DpR<40%(P=0.001).COX multivariate analysis showed that DpR is an independent factor affecting PFS and OS,and distant lymph node metastasis is an independent factor affecting OS.The overall tolerance to treatment of the patients was good,with no grade 4 or above treatment-related adverse reactions or death.Conclusions:ETS and DpR may be predictive indicators of the efficacy and prognosis of PD-1 inhibitors combined with trastuzumab and first-line chemotherapy for HER-2 positive advanced gastric cancer.

Objective: To explore the clinical effect of free anterolateral thigh flap in repairing large annular soft tissue defect of lower leg after burn. Methods: From January 2014 to December 2018, 9 patients with large annular soft tissue defects of lower legs after burns were hospitalized in Zhengzhou First People′s Hospital, including 1 case with wounds on both legs. After debridement, area of wounds was 16 cm×11 cm-38 cm×21 cm, and the burn wounds were repaired with free anterolateral thigh flaps in the area of 18 cm×12 cm-32 cm×24 cm. End-to-end anastomosis of posterior tibial vessels or anterior tibial vessels with lateral circumflex femoral vessels was performed in manual way or by microvascular stapler. For the affected legs without condition for anastomosis, the sound medial lower leg flaps with areas of 10 cm×8 cm-15 cm×10 cm were excised and made into skin tubes, the posterior tibial vessels of the flaps were anastomosed with the vessels of free anterolateral femoral flaps, and the wounds of the injured lower legs were repaired by bridge-type cross-over free transplantation of anterolateral thigh flaps. The pedicles were broken 4 to 5 weeks later. The donor site was transplanted with autologous intermediate split-thickness skin graft from thigh. The outcome of the treatment, the number of perforators included in the flaps, and the anastomotic vessel in the recipient area of patients were recorded. The anastomosis time between manual way and microvascular staplers was recorded and compared. The patency of blood vessels, methods of free transplantation, and follow-up condition were recorded. Data were processed with Wilcoxon rank sum test for two independent samples. Results: All the 10 free flaps and skin grafts of 9 patients survived, and all the wounds were closed by primary operation. Seven flaps contained two perforators each, and three flaps contained three perforators each. The anastomotic vessels were posterior tibial vessels in 6 recipient areas and anterior tibial vessels in 4 recipient areas. Microvascular stapler was used to anastomose 12 veins, while 8 veins and 10 arteries were anstomosed manually. The time consumed by the former method was 4.00 (3.55, 4.38) min, significantly shorter than 12.80 (12.13, 13.40) min of the latter (W=78.00, P<0.01). The patency rates of veins and arteries were 100%. There was no vascular crisis due to vascular anastomosis. Three patients underwent bridge-type cross-over free transplantation, while the others underwent conventional free transplantation. Follow-up for 3 to 30 months showed that the donor site of the thigh had good motor function, without numbness or pain, but hypertrophy of scar could be seen. Four patients had slightly overstaffed flaps transplanted in the recipient area of the lower legs, while the other patients were satisfied with their appearance, and the walking function of the affected limbs gradually recovered. Conclusions Free anterolateral thigh flap transplantation is a safe and reliable clinical limb salvage method for the repair of large annular soft tissue defect of lower leg after burn. Intraoperative application of microvascular stapler for venous anastomosis can shorten the time of vascular anastomosis and has great clinical application value.