Accuracy and Safety of Robot-Assisted Drilling Decompression for Osteonecrosis of the Femoral Head.

OBJECTIVE: To investigate the safety and superiority of robot-assisted femoral head drilling decompression in the treatment of femoral head necrosis. METHODS: A total of 63 patients who underwent borehole decompression of the femoral head in our hospital from January 2016 to March 2019 were recruited. Patients were divided into two groups for comparison according to surgical methods. In the robot-assisted surgery group, there were 30 cases with 41 femoral heads. The conventional group had 33 cases and 46 femoral heads. All patients signed the consent form before the operation. The follow-up time was 6 months. The incision lengths, operation times, intraoperative blood loss, intraoperative fluoroscopies, guide needle punctures, postoperative Harris scores, and postoperative complications of the two groups were compared. RESULTS: The incision length of the robot surgery group was 5.16 ± 0.41 cm, while that of the traditional surgery group was 7.42 ± 0.50 cm. The operation time of the robot surgery group was 46.99 ± 4.94 min, while that of the traditional surgery group was 55.01 ± 6.19 min. The fluoroscopy frequency of the robot surgery group was 10.50 ± 1.78 times, while that of the traditional surgery group was 17.91 ± 2.20 times. The intraoperative blood loss in the robotic surgery group was 20.62 ± 2.52 mL, while that in the conventional surgery group was 52.72 ± 3.39 mL. In the robot operation group, each femoral head guide needle was punctured three times, and the puncture was successful one time. The number of guided needle punctures in the traditional group was 8.02 ± 1.73. The difference between the two groups was statistically significant (P < 0.05). The Harris score was 69.53 ± 7.51 in the robot surgery group and 68.38 ± 7.26 in the traditional surgery group one month after surgery, 78.52 ± 6.49 in the robot surgery group and 76.41 ± 7.95 in the traditional surgery group three months after surgery, and 83.32 ± 8.62 in the robot surgery group and 81.74 ± 6.20 in the traditional surgery group six months after surgery. There was no significant difference between the two groups (P > 0.05). In the traditional group, there was one case of incision infection and one case of femoral head collapse during follow-up. In the robot group, there were no complications, such as incision infection and deep vein thrombosis. No collapse of the femoral head was found in the robot group during follow-up. CONCLUSION: The positioning system of the orthopaedic robot is an ideal method for the treatment of femoral head necrosis. This method has the advantages of simple operation, accurate drilling, a short operation time, less surgical trauma, less radioactivity, and good recovery of hip joint function.

SuperPATH Minimally Invasive Approach to Total Hip Arthroplasty of Femoral Neck Fractures in the Elderly: Preliminary Clinical Results.

OBJECTIVE: To investigate the clinical efficacy and advantages of the SuperPATH minimally invasive approach to total hip arthroplasty in the treatment of femoral neck fractures in the elderly. METHODS: From January 2016 to September 2018, 110 cases of elderly patients with femoral neck fractures were included in the present study. According to the method of operation, the patients were divided into two groups for comparison. There were 55 cases of the SuperPATH minimally invasive approach to total hip arthroplasty and 55 cases with the conventional posterolateral approach to total hip arthroplasty. The operation time, the length of incision, the amount of operative blood loss, the hospitalization time, and the hospitalization cost were compared between the two groups. The position of total hip prosthesis was observed during the follow-up period. All patients were evaluated for the degree of hip joint pain and the function of the hip joint using the visual analog score (VAS) and the Harris score at 1 week, 1 month, 3 months, 6 months, and 12 months after the operation. RESULTS: All patients were followed up for at least 12 months. The operation time was 108.58 ± 15.87 min in the SuperPATH group and 102.51 ± 19.61 min in the conventional group. The length of incision was 6.65 ± 1.53 cm in the SuperPATH group and 17.08 ± 1.40 cm in the conventional group. The amount of operative blood loss was 147.51 ± 28.84 mL in the SuperPATH group and 170.22 ± 25.34 mL in the conventional group. The hospitalization time was 10.05 ± 2.52 days in the SuperPATH group and 13.36 ± 3.39 days in the conventional group. The hospitalization cost was 6871.78 ± 141.63 dollars in the SuperPATH group and 7791.09 ± 184.88 dollars in the conventional group. Compared with the conventional group, the SuperPATH group had shorter incision length, less blood loss, shorter hospitalization time, and lower hospitalization cost. There was significant difference between the two groups (P < 0.05). In the two groups, there were no complications such as infection, lower extremity venous thrombosis, prosthesis loosening, periprosthetic fracture, and dislocation during the follow-up period. The VAS score was 4.45 ± 0.94 in the SuperPATH group and 4.89 ± 0.79 in the conventional group at 1 week after the operation. There was significant difference between the two groups (P < 0.05). The Harris score was 75.36 ± 3.36 and 80.25 ± 3.09 in the SuperPATH group and 68.80 ± 3.25 and 77.35 ± 3.77 in the conventional group at 1 week and 1 month after the operation, respectively. There was significant difference between the two groups (P < 0.05). In the analysis of the operation time, the VAS score at 1 month, 3 months, 6 months, and 12 months after the operation, and the Harris score at 3 months, 6 months, and 12 months after surgery, there was no significant difference between the two groups (P > 0.05). CONCLUSION: The SuperPATH minimally invasive approach to total hip arthroplasty is an ideal method for the treatment of femoral neck fractures in the elderly. This method has the advantages of the relatively simple operation, short incision, less blood loss, and less trauma. The patients had short hospitalization times, low hospitalization costs, and good recovery of hip joint function.

A Comparative Study of Intramedullary Nail Strengthened with Auxiliary Locking Plate or Steel Wire in the Treatment of Unstable Trochanteric Fracture of Femur.

OBJECTIVE: To compare the clinical outcomes of unstable femoral trochanteric fracture treated by proximal femoral intramedullary nail enhanced with lateral locking plate versus cerclage steel wire. METHODS: A retrospective study was conducted on 83 patients who received open reduction and internal fixation with proximal femoral intramedullary nail combined with lateral enhanced fixations for unstable femoral trochanteric fractures from March 2015 to January 2017 in our hospital. Of these patients, 39 received the lateral enhanced fixation with locking plate, while the remaining 44 had cerclage wire as additional fixation. The clinical data were compared between the two groups. RESULTS: All the patients in the study had surgical procedures performed smoothly. Although the plate group had significantly longer operation times and significantly higher hospitalization expenses than the wire group (P < 0.05), no statistically significant differences in intraoperative blood loss and hospital stay were proved between the groups (P > 0.05). The follow-up period lasted for 12-24 months with an average of (16.81 ± 2.92) months. The plate group returned to full-weight bearing significantly earlier than the wire group (P < 0.05). The Harris Hip Score (HHS) significantly increased in both groups over time postoperatively (P < 0.05). The plate group achieved a higher HHS than the wire group, which was statistically significant at 3 months (P < 0.05), whereas it became insignificant at 6 and 12 months postoperatively (P > 0.05). Regarding radiographic assessment, an excellent rate of fracture reduction was proved in 71.79% of the plate group, compared to 45.45% of the wire group, which was statistically significant (P < 0.05). In addition, fracture healing was achieved significantly earlier in the plate group than the wire group (P < 0.05); nevertheless, no statistically significant difference was noted in neck-shaft angle at the latest follow-up between the two groups (P > 0.05). At the latest follow-up, two cases of implant loosening and two cases of coxa varus were found in the plate group, while one case of femoral head necrosis and three cases of coxa varus were revealed by radiographs in the wire group. CONCLUSION: The cerclage wire has benefits of saving time and operation costs; however, the locking plate has the advantages of improving fracture reduction quality, shortening time to full weight bearing and fracture healing, and improving hip function recovery due to the lateral additional fixations to proximal femoral intramedullary nail for unstable trochanteric fractures.

Regional Injury Classification and Treatment of Open Pelvic Fractures.

OBJECTIVES: To propose the regional injury classification of open pelvic fracture and summarize the characteristics of its treatment. METHODS: Clinical data for 67 open pelvic fractures treated from January 2001 to December 2017 were retrospectively analyzed. There were 48 male and 19 female patients. The patients were aged from 12 to 68 years old, and their pelvic fractures were categorized according to Tile classification (type A: 23 cases; type B: 19 cases; type C: 25 cases). Main injury distribution: pubic perineum, 29 cases; ilioinguinal, 20 cases; and sacroiliac, 7 cases. There were 5 cases of ilioinguinal-perineum and 6 cases of sacroiliac-perineum injury. Based on the region of the injury, the mortality and combined injury of each group were observed. The relationship between regional injury groups and death was examined. RESULTS: Following active treatment, 28 patients died. The mortality rate was 41.8% (28/67), with 39 patients surviving. The average follow-up time was 6 months (3 months to 1 year after discharge). Majeed pelvic fracture score: the score was excellent in 12 cases, good in 14 cases, fair in 9 cases, and poor in 4 cases; there was an excellent and good rate of 66.7%. Open pelvic fracture regional injury classification includes: the perineal type (type I); the ilioinguinal type (type II); the sacroiliac type (type III); and the composite type (type IV). All types were independent of each other, and the mortality of open pelvic fractures was positively correlated with this classification, with a correlation coefficient of 0.620 (P = 0.001 < 0.05; the difference was statistically significant). In this study, cystourethral injury, anorectal injury, and infection were the main combined injuries of type I. The type II and III injuries were mainly iliac vascular injuries. The main combined injuries of type IV were infection, injury of ilium vessels and its branches. There was a statistical difference among the combined injuries of each subtype (P = 0.001 < 0.05). CONCLUSIONS: The criteria for regional classification were clear, the mortality of the four subtypes increased gradually, and the incidence of combined injury of each subtype varied. Each subtype had different therapeutic characteristics.

In Situ Reduction and Fixation of the Anterior Medial Fenestration Approach for Femoral Head Fracture.

OBJECTIVE: To investigate the feasibility and clinical application of the anterior medial fenestration approach in the treatment of Pipkin type I and II femoral head fractures. METHODS: The hips of two anti-corrosion adult specimens treated with formalin were dissected and the anatomical structures and directional characteristics of the anterior medial main muscles, ligaments, blood vessels, and nerves were observed. The anterior medial fenestration approach was performed on bilateral hips of four fresh frozen specimens to determine the required pulling direction of the stripped muscles and ligaments during surgery. In addition, the vascular and nerve traction protection directions exposed in the approach were observed and analyzed. The feasibility of this approach was assessed, and the operative approach and critical anatomical depth were measured. We retrospectively analyzed 12 patients with Pipkin type I and II femoral head fractures who underwent in situ reduction and fixation by anterior medial fenestration in our hospital from February 2016 to April 2018. The study group included 3 men and 9 women aged 37-59 years (mean, 48.50 years). There were 8 cases of Pipkin type I and 4 cases of Pipkin type II. The operation time, blood loss, fracture healing time, last Thompson-Epstein evaluation, and Harris score were recorded. RESULTS: A total of 8 fresh frozen specimens from 4 bilateral hips were exposed by anterior medial fenestration. The upper boundary of observation fenestration was the pubic body (anterior acetabulum), and the outer upper boundary was the iliacus and the psoas muscle. The lateral boundary was the rectus femoris and the femoral vessels, while the lower boundary was the transverse branch of the medial femoral circumflex artery and vein. The medial boundary was the pubis muscle, the short adductor muscle, and the long adductor muscle. The pubofemoral and iliofemoral ligaments were observed during fenestration. By cutting open the joint capsule and moving the hip joint, the four quadrants of the femoral head can be exposed. Twelve patients with femoral head fractures who were treated with anterior medial fenestration underwent in situ reduction and fixation. The operation time was 96.25-118.75 min (median, 100 min), and the blood loss was 115.00 ± 22.76 mL. The follow-up time was 18.58 ± 4.48 months, and the fracture healing time was 144.17 ± 14.53 days. The last Thompson-Epstein evaluation was excellent in 6 cases, good in 4 cases, and fair in 2 cases; the excellent and good rate was 83.33%. Finally, the last Harris score was 85.08 ± 5.73 points. CONCLUSIONS: The upper and lower boundaries, inner and outer boundaries, and rear anatomical structure of the anterior medial fenestration approach were defined. The movable hip joint can expose the four quadrants in front of the femoral head in this fenestration. Anterior medial fenestration in situ reduction and fixation surgery is feasible and safe for the treatment of Pipkin type I and II femoral head fractures.

Treatment of Femoral Neck Fractures with Cannulated Screw Invasive Internal Fixation Assisted by Orthopaedic Surgery Robot Positioning System.

OBJECTIVE: To investigate the clinical efficacy and advantages of cannulated screw internal fixation assisted by the orthopaedic surgery robot positioning system in the treatment of femoral neck fractures. METHODS: The clinical data of 128 patients with femoral neck fractures which had been treated with cannulated screw internal fixation from January 2016 to July 2018 were retrospectively analyzed. Among them, 63 patients were treated with cannulated screw assisted by orthopedic robot positioning system (orthopaedic surgery robot group), and 65 patients were treated with traditional cannulated screw (traditional surgery group). The operation time, number of intraoperative fluoroscopy, number of guide needle placements, and the amount of operative blood loss were compared between the two groups. The success rate of one-time nail placement and the fracture healing rate were calculated. Fracture healing and internal fixation were observed. The hip joint function was evaluated by the Harris hip score 1 year after operation. RESULTS: All patients were followed up for 12 to 24 months. The operation time was 65.70 ± 9.87 min in the robot group and 73.74 ± 9.78 min in the traditional group. The number of intraoperative fluoroscopy was 13.67 ± 4.39 times in the robot group and 17.09 ± 4.02 times in the traditional group. The number of guide needle placements was 9.95 ± 3.72 times in the robot group and 13.78 ± 4.39 times in the traditional surgery group. The success rate of one-time nail placement was 100% (63/63) in the robot group and 49.23% (32/65) in the traditional group. The amount of operative blood loss was 15.25 ± 6.21 mL in the robot group and 25.51 ± 6.97 mL in the traditional group. Compared with the traditional group, the robot group had shorter operation time, less fluoroscopy, less needle placement, less bleeding, and higher success rate of one-time nail placement. There was a significant difference between the two groups (P < 0.05). In the robot group, there was no infection, loosening of internal fixation, fracture displacement, and osteonecrosis of femoral head during the follow-up period. The fracture healing rate was 100% (63/63). In the traditional group, there were two cases of loosening of internal fixation and one case of osteonecrosis of femoral head during the follow-up period. The fracture healing rate was 100% (65/65). All patients were evaluated for hip joint function 1 year after operation. The Harris hip score in the robot group was 86.86 ± 4.74, and the Harris hip score in the traditional surgery group was 83.08 ± 5.44. Compared with the traditional group, the Harris hip score in the robot group was higher than that in the traditional group. There was significant difference between the two groups (P < 0.05). The excellent and good rate were 92.06% (58/63) in the robot group and 80% (52/65) in the traditional group. There was no significant difference between the two groups (P > 0.05). CONCLUSION: Cannulated screw internal fixation assisted by the orthopaedic surgery robot positioning system is an ideal method for the treatment of femoral neck fractures. This method has the advantages of relatively simple operation, more accurate screw placement during operation, high success rate of one-time nail placement, short operation time, less surgical trauma, less radiation, and good recovery of hip function.

Comparative Study of Percutaneous Sacroiliac Screw with or without TiRobot Assistance for Treating Pelvic Posterior Ring Fractures.

OBJECTIVES: To analyze the curative effect of TiRobot surgical robotic navigation and location system-assisted percutaneous sacroiliac screw fixation and percutaneous sacroiliac screw by traditional fluoroscopy, and to summarize the safety and benefits of TiRobot. METHODS: A total of 91 patients with pelvic posterior ring fractures from December 2015 to February 2018 were included in this study. According to the surgical methods selected by the patients, the patients were divided into a TiRobot surgical robotic navigation and location system group (TiRobot group) and a percutaneous sacroiliac screw fixation group (traditional group). Statistical indicators included the number of sacroiliac screws, the time of planning the sacroiliac screw path, fluoroscopy frequency, fluoroscopy time, operation time, length of incision, blood loss, anesthesia time, the healing process of skin incisions, and fracture healing time. Fracture reduction was evaluated according to the maximum displacement degree at the inlet and outlet view X-ray or CT. Matta standard was used to evaluate fracture reduction. At the last follow-up, the Majeed function system was used to evaluate the function. RESULTS: All patients were followed up for 8 to 32 months. A total of 66 sacroiliac screws were implanted in the TiRobot group. A total of 43 sacroiliac screws were implanted in the traditional group. There were statistically significant differences in terms of fluoroscopy frequency, fluoroscopy time, operation time, incision length, anesthesia time, and blood loss between the two groups; the TiRobot group was superior to the traditional group. The healing time of the TiRobot group and the traditional group was 4.61 ± 0.68 months (range, 3.5-6.3 months) and 4.56 ± 0.78 months (range, 3.4-6.2 months), respectively, and there was no statistical difference. Postoperatively, by Matta standard, the overall excellent and good rate of fracture reduction was 89.28% and 88.57%, respectively. At the last follow-up, by Majeed function score, the overall excellent and good rate was 91.07% and 91.43%. There was no statistical difference between the two groups. CONCLUSION: Sacroiliac screw implantation assisted by TiRobot to treat the posterior pelvic ring fractures has the characteristics of less trauma, shorter operation time, and less blood loss. TiRobot has the characteristics of high safety and accuracy and has great clinical application value.

Intramedullary Nail Fixation Assisted by Orthopaedic Robot Navigation for Intertrochanteric Fractures in Elderly Patients.

OBJECTIVE: To compare the clinical efficacy of intramedullary nail fixation for intertrochanteric fractures assisted by orthopaedic robot navigation and the traditional intramedullary nail fixation in elderly patients, and to investigate the application advantages of intramedullary nail fixation for femoral intertrochanteric fractures assisted by orthopaedic robot navigation in the elderly. METHODS: Among the 51 patients with intertrochanteric fractures who were selected from April 2015 to September 2017 in the Affiliated Hospital of Chengdu University, 25 patients underwent the intramedullary nail fixation assisted by orthopaedic robot navigation (orthopaedic robot navigation surgery group) and 26 patients underwent the traditional intramedullary nail fixation (traditional surgery group). The operation time, the number of intraoperative fluoroscopy images taken, the frequency of guide pins inserted into the femoral marrow cavity, the amount of intraoperative bleeding, and the one-time success rate of the guide pin inserted into the femoral marrow cavity were recorded. Fracture healing and internal fixation were observed. The Harris score was used to evaluate hip joint function 1 year after surgery. RESULTS: All patients were followed up for 12-24 months. The operation time was 65.44 ± 8.01 min in the orthopaedic robot navigation surgery group and 77.50 ± 16.64 min in the traditional surgery group. The number of intraoperative fluoroscopy images taken was 10.28 ± 0.61 in the orthopaedic robot navigation surgery group and 13.23 ± 1.75 in the traditional surgery group. The frequency of guide pins inserted into the femoral marrow cavity was 1.00 ± 0.00 times in the orthopaedic robot navigation surgery group and 2.46 ± 1.10 times in the traditional surgery group. The one-time success rate of intramedullary pin puncture was 100% (25/25) in the orthopaedic surgical robot navigation surgery group and 19.23% (5/26) in the traditional surgery group. The amount of surgical bleeding was 90.80 ± 14.98 mL in the orthopaedic robot navigation surgery group and 118.46 ± 32.21 mL in the traditional surgery group. Compared with the traditional surgery group, the operation time of the orthopaedic surgical robot navigation surgery group was shorter (P < 0.05), the number of intraoperative fluoroscopy images taken was fewer (P < 0.05), the frequency of guide pins inserted into the femoral marrow cavity was lower (P < 0.05), the one-time success rate of intramedullary pin puncture was higher (P < 0.05), and the amount of surgical bleeding was less (P < 0.05). One year after surgery, fracture healing occurred in both groups without failure of internal fixation or fracture displacement. The Harris score of hip function in the orthopaedic robot navigation surgery group was 86.68 ± 6.23 and that in the traditional surgery group was 82.69 ± 6.85. It was higher than that in the traditional surgery group (P < 0.05). The fine rate of hip joint function in the orthopaedic robot navigation surgery group was 84.00% (21/25) and that in the traditional surgery group was 73.07% (19/26). There was no significant difference between the two groups (P > 0.05). CONCLUSION: Intramedullary nail fixation for intertrochanteric fractures assisted by orthopaedic robot navigation in elderly patients is an ideal method, offering a short operation time, minimal surgical trauma, less radiation, and good recovery of hip function.

Sodium intake, salt taste and gastric cancer risk according to Helicobacter pylori infection, smoking, histological type and tumor site in China.

AIM: The risk factors mostly strongly associated with gastric cancer are gastric bacteria Helicobacter pylori and diet. Using a case-control study among residents in Jinan, we examined the association between the salt taste and gastric cancer according to H. pylori infection, smoking and histological type as well as tumor site. METHODS: This population-based case-control study included 207 cases and 410 controls. Data on potential risk factors of gastric cancer were obtained by interview of cases and controls with a questionnaire, salt taste preference was measured for all subjects, and IgG antibodies to H. pylori were applied to assess infection. Risk measures were determined using unconditional logistic regression. RESULTS: The proportions of salt taste at intervals of 1.8-7.2 g/L and ≥ 7.2 g/L were significantly higher in cases than controls, with ORs of 1.56 (1.23-3.64) and 2.03 (2.12- 4.11), respectively, subjects with high salt intake having an elevated risk for gastric cancer when infected with H. pylori. Significant modification by smoking and tumor site was observed across the different measures of salt intake, the highest salt taste showed higher cancer risk in ever smokers or with non-cardia cancers. CONCLUSION: Our study supports the view that high intake of sodium is an important dietary risk factor for gastric cancer, with a synergistic effect found between salt and H.pylori and smoking, dependent on the tumor site.

Abnormal expression of NRF-2α in hepatocellular carcinoma identified with a newly prepared monoclonal antibody against human NRF-2α protein.

Human nuclear respiratory factor 2 alpha subunit (NRF-2α) is fundamentally important to cell function and the development. We aimed to establish the monoclonal antibody (MAb) against the human NRF-2α protein and to investigate its distribution in human hepatocellular carcinoma (HCC) and tumor-adjacent tissues. The 6× His-NRF-2α fusion protein was successfully induced and purified. One monoclonal antibody (MAb) against human NRF-2α, 1-D10-E1-B11-G3 (IgG1), effective in detecting the recombinant and the cellular protein, was characterized. Using immunohistochemical analysis, the expression of NRF-2α was investigated in 38 cases of HCC specimens and 14 cases of tumor-adjacent specimens. Staining was found positive in 9 cases of HCC tissues (23.7%) and 8 cases of normal hepatic tissues (57.1%). The higher-grade frequency of expression of NRF-2α in tumor-adjacent tissues was significantly higher (P < 0.01) than that in tumor tissues, suggesting that NRF-2α may play important roles in carcinogenesis of HCC.

[Significant increase of glucose transport activity in breast cancer].

OBJECTIVE: To study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma. METHODS: A total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: In normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA). CONCLUSIONS: Glucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Heat shock protein 70 fused to or complexed with hantavirus nucleocapsid protein significantly enhances specific humoral and cellular immune responses in C57BL/6 mice.

Heat shock proteins (HSPs) are known to act as an effective molecular adjuvant to enhance the induction of antigen peptide-specific cellular immunity, when coupled with the antigen or peptide. Hantaan virus (HTNV) nucleocapsid protein (NP) is relatively conserved among hantaviruses and highly immunogenic in both animals and humans. To analyze the influence of HSP70 on NP vaccine potency, and evaluate the possibility of developing a novel effective vaccine against hantaviruses, we constructed prokaryotic expression plasmids, and expressed three recombinant proteins, namely, HTNV NP, HSP70 and HSP70-NP fusion protein. As an alternative to fusion protein, we also generated HSP70 and HTNV NP complexes (HSP70+NP) in vitro. C57BL/6 mice were immunized with those recombinant proteins, the humoral and cellular responses elicited against NP were measured by ELISA, fluorescence flow cytometry, cytotoxicity assays, and IFN-gamma ELISPOT assay. We found that immunization of mice with HSP70-NP fusion protein, or HSP70+NP complexes elicited significantly higher NP-specific antibody titers, frequencies of IFN-gamma-producing cells and cytotoxic T lymphocyte (CTL) activities in vivo than conventional HTNV NP vaccination. Antibody isotype analysis showed that the antibody response was characterized by a higher HTNV NP-specific titer of IgG2a than IgG1 antibodies, resulting in a significant higher IgG2a/IgG1 ratio. By comparison, HSP70-NP fusion protein is significantly superior to HSP70+NP complexes in enhancement of NP antigenicity. These results indicated that HSP70, when fused to or complexed with HTNV NP, greatly enhance NP vaccine potency by preferential induction of a predominant Th1 immune response in a NP-specific, HSP70-dependent manner.

A survivin double point mutant has potent inhibitory effect on the growth of hepatocellular cancer cells.

Survivin is an attractive target in cancer therapy. Previous studies have demonstrated that survivin dominant-negative mutants T34A and C84A were able to induce apoptosis in cancer cells. Given that they had different mechanisms in inducing apoptosis, our study was undertaken to determine whether a survivin double point mutant (TC34,84AA) could achieve more potent inhibitory effect on the growth of hepatocellular cancer cells. Adenoviruses expressing survivin mutants were constructed and transduced into hepatocellular cancer cells. The inhibitory effect of the survivin mutants on cancer cell growth was measured. Transduction of cancer cells with all three survivin mutants resulted in significant apoptosis. Compared with survivin mutants T34A or C84A alone, the cancer killing effect of survivin TC34,84AA was much stronger. In addition, the survivin mutants were more sensitive than wild type survivin to the degradation in the ubiquitin-proteasome pathway. Our results suggest that adenovirus-delivered dominant-negative survivin TC34,84AA promotes apoptosis-mediated hepatocellular carcinoma suppression, and could potentially be a promising candidate for cancer therapies.

HSP70 gene fused with Hantavirus S segment DNA significantly enhances the DNA vaccine potency against hantaviral nucleocapsid protein in vivo.

Heat shock proteins (HSPs) have been shown to act as adjuvants when coadministered with peptide antigens or given as fusion proteins and enhance the vaccination efficiency. To evaluate the enhancement of the potency of Hantaan virus (HTNV) nucleocapsid protein (NP) immunogenicity by heat shock protein 70 (HSP70), we developed a novel chimeric HTNV S-HSP70 DNA vaccine plasmid by genetically linking HSP70 gene to the full-length HTNV S segment DNA (HTNV S DNA). C57BL/6 mice were immunized with this plasmid followed by a subsequent boost with homologous recombinant protein. The levels of HTNV NP-specific antibody and cellular immune response were measured by use of ELISA, fluorescence activated cell sorter (FACS) analysis, cytotoxicity assay, and IFN-gamma ELISPOT assay. We found that HTNV S-HSP70 DNA vaccination significantly increased the levels of HTNV NP-specific antibody, IgG2a/IgG1 ratio, IFN-gamma producing CD8+ T-cell precursor frequencies, and cytotoxic T lymphocyte (CTL) response when compared with immunization with HTNV S DNA alone or HTNV S DNA physically mixed with HSP70 DNA. By contrast, HSP70 DNA or vector DNA immunization could not induce appreciable levels of specific antibodies and CTL response. Thus, we demonstrate for the first time that HSP70-based HTNV S DNA can induce both humoral and cellular immune response specific for HTNV NP and is a promising candidate DNA vaccine for HTNV infection.

[Construction of glucosylceramide synthase-specific siRNA expression vector and its efficiency in reversal of drug resistance in breast carcinoma cells].

OBJECTIVE: To construct a glucosylceramide synthase (GCS)-specific small interfering RNA (siRNA) expression vector and to investigate the inhibitory effect of this siRNA on GCS expression and drug resistance in breast carcinoma cells. METHODS: Two GCS gene-specific siRNAs were designed and cloned into the expression vector pSUPER to generate the plasmids pSUPER-GCS1 and pSUPER-GCS2. Human adriamycin (ADM)-resistant breast carcinoma cells of the line MCF-7/ADR and human adriamycin-sensitive breast carcinoma cells of the line MCF-7 were cultured and transfected with pSUPER-GCS1, pSUPER-GCS2, and blank vector pSUPER as controls. The expression of GCS mRNA was assayed by RT-PCR and the expression of GCS protein was observed by flow cytometry. The 50% inhibition concentration of ADM on MCF-7/ADR cells was evaluated by MTT method. Flow cytometry was performed to determine the ratio of apoptosis. RESULTS: Double enzyme digestion analysis and DNA sequencing confirmed that pSUPER-GCS1 and pSUPER-GCS2 were successfully constructed. The GCS protein positive rate of the MCF-7/ADR cells 48 hours after transfection with pSUPER-GCS1 and pSUPER-GCS2 were 8.3% +/- 1.0% and 9.2% +/- 0.8% respectively, significantly lower than that before transfection (68.3% +/- 0.6%), with a inhibition rate of 89.4% and 88.5% respectively (both P < 0.01). Forty-eight hours after transfection with pSUPER-GCS1 and pSUPER-GCS2, the relative reversal rates of sensitivity to ADM of the MCF-7/ADR cells were 93.7% and 91.6%. Flow cytometry showed that the apoptotic rate of the MCF-7/ADR cells was 0.80 +/- 0.06 before transfection, 15.38 +/- 1.16 after transfection with pSUPER-GCS1 and 13.92 +/- 1.73 after transfection with pSUPER-GCS2 (both P < 0.05), and was 0.87 +/- 0.12 in the cells transfected with blank vector (P > 0.05). CONCLUSION: A GCS-specific small interfering RNA expression vector has been constructed successfully that suppresses the GCS expression and reverses the multidrug resistance in breast carcinoma cells by increasing the ratio of apoptosis in drug-resistant cells.

[Expression and its significance of b-FGF in human benign prostatic hyperplasia and prostatic carcinoma tissues].

AIM: To investigate the clinical significance of basic fibroblast growth factor (b-FGF) in human benign prostatic hyperplasia (BPH) and prostatic carcinoma (Pca). METHODS: b-FGF expressions were examined by RNA dot blot hybridization and immunohistochemical staining in prostate tissues from 40 normal cases (NP), 38 BPH and 36 Pca. RESULTS: The expressions of b-FGF in BPH and Pca tissues were significantly higher than those in normal prostate tissues (P < 0.01). There was no significant correlation between the increased expression of b-FGF and the graduation of BPH, pathologic grading as well as clinical stages of Pca. CONCLUSION: b-FGF may be secreted from BPH and Pca tissues themselves, which is closely involved in the genesis and development of BPH and Pca.