Objective:To investigate the clinical efficacy of 3D printed metal augment or tibial prosthesis for reconstruction of large bone defects in total knee arthroplasty (TKA) and knee revision surgery.Methods:A total of 7 patients (7 knees) with TKA or knee revision who were admitted to the Department of Orthopaedics of the Second Affiliated Hospital of Zhejiang University School of Medicine with large bone defects from July 2018 to December 2023 were retrospectively analyzed, including 4 patients with TKA and 3 patients with knee revision. There were 3 males and 4 females, aged 58.7±7.6 years (range, 54-68 years), 3 patients with left knee and 4 patients with right knee. All the patients had bone defects in the knee joint (AORI type III), 2 cases had bone defects only in the femur, 4 cases had bone defects only in the tibia, and 1 case had bone defects in both the tibia and femur, which were treated with personalized reconstruction using 3D printing. Hip-knee-ankle angles, American Knee Society score (KSS) before and after surgery were compared, and postoperative complications were observed.Results:All patients successfully completed the operation, and the operation time was 189.3±35.5 min (range, 125-240 min). Complex TKA was performed in 4 cases with surgical times of 175, 195, 210, and 240 min, and revision surgery was performed in 3 cases with surgical times of 125, 180, and 200 min, respectively. Intraoperative blood loss was 114±24.4 ml (range, 100-150 ml). Five cases used 3D printed metal augment, and two used 3D printed one-piece tibial components. All patients were followed up for 2, 2, 5, 6, 7, 20, 57 months, respectively. The KSS of the five patients at 3 months postoperatively were 56, 61, 66, 56, and 56 points, respectively, greater than the preoperative scores of 35, 44, 36, 27, and 41 points. The KSS functional scores of the five patients at 3 months postoperatively were 45, 45, 45, 30, and 45 points, respectively, which were greater than the preoperative scores of 30, 30, 15, 20, and 20 points. The hip-knee-ankle angle was 181.8°±3.4° (range, 177.9° to 188.0°) at the final follow-up and 175.8°±12.4° (range, 153.3° to 192.1°) before surgery, with no significant difference ( t=-1.230, P=0.242). At the final follow-up, the 3D printed component was well integrated with the bone surface, the prosthesis was securely positioned, and the force lines of the lower limbs were normal. There were no postoperative complications such as poor wound healing, infection, fat liquefaction, nerve injury, deep vein thrombosis of lower limbs, knee joint stiffness, periprosthesis infection and loosening. Conclusion:Using 3D printed metal augment or tibial prosthesis to reconstruct the huge bone defect in TKA and revision has a satisfactory early clinical effect, satisfactory joint function and good surgical safety.

Objective:To analyze the epidemiological and etiological characteristics of a dengue fever outbreak in Hunan province in 2018.Methods:Real-time PCR assay was performed for the laboratory diagnosis of 8 suspected dengue fever cases. Etiological surveillance was performed in 186 suspected dengue fever cases and fever cases who had close contacts with dengue fever patients. C6/36 cells was used for the virus isolation from acute phase serum. By sequencing the full length of E genes of 15 dengue virus strains, phylogenetic analysis was performed based on the sequences obtained, including reference sequences from the NCBI GenBank database, the serotypes and gene subtypes of the virus were analyzed to trace the possible source of transmission. An emergency monitoring of vector density and a retrospective survey of sero-epidemiology in healthy population were conducted in the epidemic area.Results:In the serum samples of 8 suspected patients, 6 were dengue virus RNA positive, and 4 were NS1 antigen positive. In 186 suspected patients, 96 were dengue virus nucleic acid, NS1 antigen or antibody positive in etiological test. A total of 64 dengue virus strains were isolated. The phylogenetic analysis showed that all the dengue virus strains belonged to type 2, which might be from Guangdong or Zhejiang provinces. The Bretub index was up to 65, indicating an extremely high risk of transmission. The positive rate of the dengue virus IgG antibody was 0.53%(2/377) in retrospective survey of 377 healthy people.Conclusion:The field epidemiologic and the molecular genetics analyses showed the outbreak of dengue fever in Hunan in 2018 was caused by imported cases and dengue virus 2.

Objective:To characterize the clinical and epidemiological changes, treatments, and prognoses of primary esophageal small cell carcinoma (PESC). Methods:A retrospective analysis was conducted using the clinical epidemiology data of 529 PESC patients se-lected from the clinical databases of 500,000 esophageal and gastric cardiac carcinomas of the Henan Key Laboratory for Esophageal Cancer Research (1992-2015). Among these patients, 241 cases were included in the survival analysis. The five-year survival rate was calculated using Kaplan-Meier analysis, and the differences in survival rates were compared using the Log-rank analysis model. Re-sults:All 529 PESC cases were analyzed, which accounted for 0.2%of esophageal cancers diagnosed in the same period. The incidence of PESC increased annually (R2=0.574). The survival rates for 1-, 2-, 3-, and 5-year of 241 PESC patients were 55%, 40%, 29%, and 9%, respectively, and the median survival time was 21.9 months. On the basis of the VALSG criteria of lung small cell carcinoma, a statisti-cal difference was observed in the overall survival rates for limited and extensive diseases (P=0.003), with the median survival time of 24.3 and 17.5 months, respectively. Furthermore, significant differences were observed on survival with various treatment modalities (P=0.004). The median survival time of PESC patients treated with combined surgery and radiochemotherapies (28.8 months) was lon-ger than those with either chemotherapy (17.8 months, P=0.015) or chemoradiotherapy (14.5 months, P=0.004). In limited disease pa-tients, the median survival time was longer in patients treated with surgery (27.7 months) than in those without surgery (16.2 months, P=0.007). Notably, the biopsy diagnosis before surgery for PESC was only 40.8%. Conclusion:PESC is a rare malignant carcinoma with increasing incidence. PESC presents poor prognosis, and the survival rate can be improved through combined therapies based on sur-gery. A high misdiagnosis rate for PESC is observed before surgery with biopsy.

OBJECTIVETo inhibit the proliferation and metastasis of colon cancer cells by increasing the expression level of B-cell translocation gene-2 (BTG2).

METHODSWestern blot assay was used to detect the expression level of BTG2 protein in the normal intestinal epithelial HIEC cells and three colon cancer cell lines SW620, HT-29 and LS174T. The expression of BTG2 protein in normal colonic epithelial tissue, colon adenoma and colon cancer tissue was detected by immunohistochemistry. The plasmid with BTG2 gene full-length sequence was transfected into colon cancer SW620 cells, and the expression of BTG2 protein was detected by Western blot. The cell growth curve was drawn by MTT test. The Ki-67-positive rate was calculated using immunofluorescence staining. The cell migration of colon cancer cells was detected by scratch test and Transwell double chamber culture system, and the pseudopodia growth of tumor cells was detected by Matrigel 3D culture system.

RESULTSWestern blot results showed that BTG2 relative expression levels were 0.83 ± 0.12, 0.18 ± 0.04, 0.20 ± 0.05 and 0.36 ± 0.07 in normal human intestinal epithelial cells HIEC, and human colon cancer cell line SW620, HT-29 and LS174T, respectively. The results of immunohistochemistry showed that the positive expression of BTG2 protein in normal colorectal tissue, colorectal adenoma and colorectal carcinoma tissues were 82.5% (33/40), 77.5%(31/40) and 17.5% (7/40), respectively, with a significant difference between two groups (P < 0.05). Immunofluorescence results showed that the positive rate of Ki-67 in the control group, empty vector group and BTG2 transfection group was (76.2 ± 8.0)%, (81.4 ± 9.7)% and (50.1 ± 7.1)%, respectively, showing a significant difference between two groups (P < 0.05). The scratch test results showed that in the control group, empty vector group and BTG2 transfection group, the distance of SW620 cells between two sides was (79.27 ± 11.24) µm, (80.65 ± 12.17) µm and (124.77 ± 19.63) µm, respectively, with a significant difference between two groups (P < 0.05). Transwell results showed that in the control group, empty plasmid group and BTG2 transfection group, the SW620 cell migration rate was (78.5 ± 13.1)%, (73.2 ± 12.9)% and (47.4 ± 9.1)%, respectively, showing a significant difference between two groups (P < 0.05). The number of neurospheres of BTG2 transfection group was decreased SW620, which had poor ductility.

CONCLUSIONSBTG2 gene is involved in colon cancer cell proliferation and metastasis, and effectively restores the function of BTG2 protein. Therefore, it may be expected to become a new option in gene therapy for colon cancer.

B-Lymphocytes ; physiology ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; genetics ; Colonic Neoplasms ; Genetic Vectors ; Humans ; Immediate-Early Proteins ; genetics ; Immunohistochemistry ; Plasmids ; Transfection ; Tumor Suppressor Proteins ; genetics

Objective To inhibit the proliferation and metastasis of colon cancer cells by increasing the expression level of B-cell translocation gene-2 (BTG2).Methods Western blot assay was used to detect the expression level of BTG2 protein in the normal intestinal epithelial HIEC cells and three colon cancer cell lines SW620, HT-29 and LS174T.The expression of BTG2 protein in normal colonic epithelial tissue, colon adenoma and colon cancer tissue was detected by immunohistochemistry.The plasmid with BTG2 gene full-length sequence was transfected into colon cancer SW620 cells, and the expression of BTG2 protein was detected by Western blot.The cell growth curve was drawn by MTT test.The Ki-67-positive rate was calculated using immunofluorescence staining.The cell migration of colon cancer cells was detected by scratch test and Transwell double chamber culture system, and the pseudopodia growth of tumor cells was detected by Matrigel 3D culture system.Results Western blot results showed that BTG2 relative expression levels were 0.83±0.12,0.18±0.04, 0.20±0.05 and 0.36±0.07 in normal human intestinal epithelial cells HIEC, and human colon cancer cell line SW620, HT-29 and LS174T, respectively.The results of immunohistochemistry showed that the positive expression of BTG2 protein in normal colorectal tissue, colorectal adenoma and colorectal carcinoma tissues were 82.5%(33/40), 77.5%(31/40) and 17.5%(7/40), respectively, with a significant difference between two groups (P<0.05).Immunofluorescence results showed that the positive rate of Ki-67 in the control group, empty vector group and BTG2 transfection group was (76.2± 8.0)%, (81.4±9.7)%and (50.1±7.1)%, respectively, showing a significant difference between two groups ( P<0.05 ) .The scratch test results showed that in the control group, empty vector group and BTG2 transfection group , the distance of SW620 cells between two sides was ( 79.27 ±11.24) μm, ( 80.65 ± 12.17) μm and (124.77±19.63) μm, respectively, with a significant difference between two groups ( P<0.05) .Transwell results showed that in the control group, empty plasmid group and BTG2 transfection group, the SW620 cell migration rate was (78.5±13.1)%, (73.2±12.9)%and (47.4±9.1)%,respectively, showing a significant difference between two groups ( P<0.05) .The number of neurospheres of BTG2 transfection group was decreased SW620, which had poor ductility.Conclusion s BTG2 gene is involved in colon cancer cell proliferation and metastasis , and effectively restores the function of BTG2 protein.Therefore, it may be expected to become a new option in gene therapy for colon cancer.

Objective To inhibit the proliferation and metastasis of colon cancer cells by increasing the expression level of B-cell translocation gene-2 (BTG2).Methods Western blot assay was used to detect the expression level of BTG2 protein in the normal intestinal epithelial HIEC cells and three colon cancer cell lines SW620, HT-29 and LS174T.The expression of BTG2 protein in normal colonic epithelial tissue, colon adenoma and colon cancer tissue was detected by immunohistochemistry.The plasmid with BTG2 gene full-length sequence was transfected into colon cancer SW620 cells, and the expression of BTG2 protein was detected by Western blot.The cell growth curve was drawn by MTT test.The Ki-67-positive rate was calculated using immunofluorescence staining.The cell migration of colon cancer cells was detected by scratch test and Transwell double chamber culture system, and the pseudopodia growth of tumor cells was detected by Matrigel 3D culture system.Results Western blot results showed that BTG2 relative expression levels were 0.83±0.12,0.18±0.04, 0.20±0.05 and 0.36±0.07 in normal human intestinal epithelial cells HIEC, and human colon cancer cell line SW620, HT-29 and LS174T, respectively.The results of immunohistochemistry showed that the positive expression of BTG2 protein in normal colorectal tissue, colorectal adenoma and colorectal carcinoma tissues were 82.5%(33/40), 77.5%(31/40) and 17.5%(7/40), respectively, with a significant difference between two groups (P<0.05).Immunofluorescence results showed that the positive rate of Ki-67 in the control group, empty vector group and BTG2 transfection group was (76.2± 8.0)%, (81.4±9.7)%and (50.1±7.1)%, respectively, showing a significant difference between two groups ( P<0.05 ) .The scratch test results showed that in the control group, empty vector group and BTG2 transfection group , the distance of SW620 cells between two sides was ( 79.27 ±11.24) μm, ( 80.65 ± 12.17) μm and (124.77±19.63) μm, respectively, with a significant difference between two groups ( P<0.05) .Transwell results showed that in the control group, empty plasmid group and BTG2 transfection group, the SW620 cell migration rate was (78.5±13.1)%, (73.2±12.9)%and (47.4±9.1)%,respectively, showing a significant difference between two groups ( P<0.05) .The number of neurospheres of BTG2 transfection group was decreased SW620, which had poor ductility.Conclusion s BTG2 gene is involved in colon cancer cell proliferation and metastasis , and effectively restores the function of BTG2 protein.Therefore, it may be expected to become a new option in gene therapy for colon cancer.

Objective To explore the post training needs,training content,training mode and the advantages and disadvantages of the existing training from the general practitioner perspective,in order to provide reasonable suggestions for the management institutions to improve the quality and effect of the post training for all general practitioners.Methods In October,2013 a questionnaire survey was conducted to 197 family physicians for their post training needs,training content and training mode.According to the interview outline,an in-depth individual interview was conducted to 20 leaders and general practitioners from the community health service center in Beijing Xicheng District on the advantages and disadvantages of general medical training.All survey information databases were established using FoxPro 6.0 software and double entry.Package was statistically analyzed with SPSS 19.0 statistical software,qualitative data using rate to represent; interviews with text descriptions,coding classification analysis.Results More than 70％ of general practitioners had the demand for strengthening the training of the clinical medicine,the knowledge of humanities and social sciences as well as clinical research methods,but the demand for the training of general medical theory and basic medicine was small,only accounting for 22.50％ general practitioners.More than 90.00％ of general practitioners were in favor of targeted flexible training methods.More than 85.00％ of general practitioners believed that existing training teachers' teaching abilities were very good,and they agreed to open general practitioner training project in medical colleges and universities.The surveyed physicians also made some criticism about too many training departments,low training quality,too many training programs,and training content repetition etc..Conclusion According to the general medical post training needs,human and scientific research of targeted training should be increased by using case discussion,site visit and many flexible and diverse teaching methods and various departments should also be coordinated and the training incentive and evaluation mechanism should be improved to guarantee the quality of training.

Objective To study the results of functional hemispherectomy for pediatric intractable epilepsy with hemisphere lesions , and to explore the indications of functional hemispherectomy , as well as the prognosis. Methods Twenty-four children with intractable epilepsy caused by hemisphere lesions were included , who underwent functional hemispherectomy. All patients were followed up for 3 ~ 48 months. Results All 24 cases obtained improvement of abnormal behavior , and no significant exacerbation of neurological deficits was observed. During the follow-up, 18 cases were in EngelⅠ(75%), 5 in Engel Ⅱ(21%), and 1 in Engel Ⅰ(4%) respectively. No long-term complication was observed. Conclusion Comprehensive presurgical evaluation is a prerequisite for the surgical treatment of pediatric intractable epilepsy caused by hemisphere lesions. Functional hemispherectomy can effectively control seizures and obviously improve the prognosis.

Objective To explore the relationship between cervical dysplasia and high risk type of Human Papillomavirus (HR-HPV),Ureaplasma urealyticum/ Mycoplasma hominis (UU/Mh) or Chlamydia Trachomatis (CT) infections with cervical epithelial DNA Ploidy Analysis,so that combining the infection with the result of cervical cancer screening test to evaluate the real risk of cancer or precancerosis to avoid over-diagnosis and over-treatment.Methods Cervical samples from 265 women on child-bearing age were involved in cervical cancer screening with DNA-Ploidy Analysis (DNA-ICM).The women' s samples also be tested by HPV test,UU/Mh test and CT test.The biopsy result is regarded to be the golden standard.Results Among the 285 biopsy results,the chronic cervicitis occupied 179,CIN I 60,CIN Ⅱ 20 CIN Ⅲ 25 and cervical cancer 1.Regression result indicated that DNA-Aneuploid and HR HPV have highest risk to cervical lesion.In the group with infectious factors,the ones with dual-factor infection in CIN Ⅰ is 32.43％ while 56.00％ in CIN Ⅲ as the most.Nevertheless,the most in CIN Ⅱ is HR-HPV as 50.00％,which emerged as 40.00％ in CIN Ⅲ.Conclusion The group with DNA aneuploidy combined with HR-HPV infection or HR-HPV and UU/Mh infection has the high potential risk of malignancy.