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Objective: To investigate the clinical characteristics and drug susceptibility test (DST) of patients infected with different nontuberculous mycobacteria (NTM). Methods: The patients with nontuberculous mycobacterial lung disease (NMLD) in Shanghai Pulmonary Hospital from March 2014 to March 2015 were studied retrospectively by analyzing the clinical characteristics, radiological features and DST results. A total of 201 NMLD patients [male 108, age(58±15) yrs] were enrolled into this study including 48 cases of M. Kansasii [male 13, age (52±16) yrs],46 cases of M. Abscess [male 46, age (57±16) yrs], 92 cases of M. Intracellulare [male 43, age (61±13) yrs], and 15 cases of M. Avium [male 6, age (67±10) yrs]. Clinical data were collected when the diagnosis was made and Chi-square test was used to compare the differences among 4 groups of patients. Bonferroni method was used for further pairwise comparisons. Results: There were significant differences among the 4 groups in the age(χ(2)=6.42, P< 0.001) and the gender(χ(2)=49.18, P< 0.001) of the patients. The history of bronchiectasis in the groups of M. Kansasii, M. Abscess , M. Intracellulare and M. Avium were 2/48, 31/46, 39/92 and 4/15 cases respectively(χ(2)=41.84, P< 0.001). For the Gamma-interferon release assays (ELISA) (IGRA), the positive rate of IGRA in the groups of M. Kansasii, M. Abscess, M. Intracellulare and M. Avium were 83%(40/48), 30%(14/46), 23%(21/92) and 33% (5/15) respectively(χ(2)=50.96, P< 0.001). The radiological features were significantly different in tree-in-bud(8/48, 35/46, 36/92 and 4/15 cases respectively, χ(2)=36.48, P< 0.001), pleural thickness or mild effusion (21/48, 36/46, 69/92 and 7/15 cases, χ(2)=19.54, P< 0.001), bronchiectasis (20/48, 39/46, 78/92 and 10/15 cases, P< 0.001) and cavities (38/48, 21/46, 63/92 and 10/15 cases, χ(2)=12.38, P< 0.001) among the 4 groups( M. Kansasii, M. Abscess, M. Intracellulare and M. Avium ). The drug resistance rates of M. Kansasii to rifampin, ethambutanol and ofloxacin were 10%(5/48), 8%(4/48) and 15%(7/48) respectively; the resistance rates of M.Intracellulare to ethambutanol was 45%(41/92), and the resistance rates of M.Abscess were all over 80% to all anti-TB drugs. The results of pairwise comparisons showed that the male proportion(46/48) and IGRAs positive rate(40/48) of patients with M. Kansasii were higher than those of other groups, and the incidence of bronchiectasis(20/48) and pleural changes(21/48) was lower than those of other groups. The female ratio(33/46), history of bronchiectasis (31/46) and tree-in-bud sign of patients(35/46) with M. Abscess were higher than those of other groups. Conclusions: There were differences in the clinical manifestations and imaging features of 4 common NMLD diseases, which were helpful for clinical differentiation. The patients with M. Kansasii infection were mainly male, with a high IGRA positive rate and fewer lesions of bronchiectasis or pleural changes. Most of the patients with M. Abscess were female, with a previous history of bronchiectasis, and with most of the lesions showing tree-in-bud signs. The NTM species had a high rate of resistance to anti-TB drugs except M. Kansasii.

Objective: To investigate the protection effects of the activation of NMDAR1(NMDA receptor 1)/ERK1/2 signal pathway on visual cortex nerve cells induced by levodopa in amblyopia rats. Methods: SD rats of SPF grade, were randomly divided into for groups of 9, group A. Control, B. MD group, C. L-levodopa(20 mg·kg(-1))+MD group, D. H-levodopa (80 mg·kg(-1))+MD group, E. H-levodopa+MD+MK801 group, F. H-levodopa+MD+PD98059 group, G. MD+MK801 group, H. MD+PD98059 group, I. MD+DMSO group. Amblyopia rats were made by suture of the right eye. Levodopa was used to treatment amblyopia by gavage, and intervened by intracerebroventricular injection of MK801 and PD98059 respectively. The expression of NR1, p-ERK1/2, ERK1/2, NGF and c-FOS were detected by Western blotting. Nissl staining was used to detect morphological changes of neurons. Neuronal apoptosis was detected by TUNEL method, and detected the expression of Caspase-3, NGF and c-FOS by immunohistochemical staining. One/Two way Chi-square analysis was used for data analysis. LSD- t test was used as comparison between every two groups. Results: The morphology of Nissl bodies in neurons was complete and clear in A group, and the size of Nissl bodies got smaller, and caused karyopyknosis and loss of neurons in visual cortex of B group. Compared with A group, the apoptosis of visual cortical neurons(23.09±2.00 vs . 2.20±0.35, t= 12.120, P= 0.000) and the number of Caspase-3 postive cells (22.70±1.50 vs . 3.30±0.54, t= 12.120, P= 0.000)were significantly increased, the expression of NGF(0.31±0.04 vs . 0.74±0.09, t= 7.674, P= 0.000) and c-FOS(0.25±0.03 vs . 0.57±0.07, t= 5.919, P= 0.000) and the rats of NGF(8.30±0.82 vs . 35.18±2.01, t= 12.37, P= 0.0000) and c-FOS (10.84±1.02 vs . 35.68±2.55, t= 9.056, P= 0.0001) postive cell were decreased significantly in B group. After treatment with levodopa, the morphology of neurons recovered, the apoptosis of visual cortical neurons relieved, the expression of NR1(0.75±0.09 vs . 0.40±0.05, t= 8.528, P= 0.001) and p-ERK1/2(2.13±0.26 vs . 0.68±0.17, t= 3.488, P= 0.008) were increased significantly, and the rats of NGF (18.07±0.87 vs . 8.30±0.82, t= 8.18, P= 0.0000) and c-FOS (19.78±0.91 vs. 10.84±1.02, t= 6.543, P= 0.0001) postive cells were significantly increased. MK-801 or PD98059 intervention could effectively attenuate the effect of levodopa. It could effectively down-regulated the expression of NR1 (0.53±0.06 vs . 0.95±0.12, t= 5.647, P= 0.005) and p-ERK1/2(1.52±0.18 vs . 2.58±0.30, t= 3.091, P= 0.013) interference with MK801 or PD98059 in MD rats. MK-801 or PD98059 intervention further promote the Nissl body volume reduced, neurons karyopyknosis, the apoptosis of visual cortical neurons and Caspase-3 expression, and restrain the expression of NGF and c-FOS. Conclusion: Levodopa played a protective role in visual cortex nerve cells of amblyopia rats at least partially through activation of NMDA-ERK1/2 signal pathway. (Chin J Ophthalmol, 2017, 53: 931-940) .

Objective: To explore the value of employing the small intestinal feeding tube in treating high position intestinal obstruction of newborn infant., Method: Five newborn infants (3 males and 2 females; 1 premature infant and 4 fully-mature infants; 2 had membranous atresia of duodenum, 1 had annular pancreas, and 2 had proximal small intestine atresia; 1 infant had malrotation). The duodenal membrane-like atresia and the blind-end of small intestine were removed and intestinal anastomosis was performed, which was combined with intestinal malrotation removal. Before the intestinal anastomosis surgery, the anesthetist inserted via nose a 6Fr small intestinal ED tube, made by CREATE MEDIC CO LTD of Japan[, Registration Number: the State Food and Drug Administration-instrument (Im.) 2007-NO.2661620]. Twenty-four hours after surgery, abdominal X-ray plain film was taken and patients were fed with syrup; 48 hours later, formula milk was pumped or lactose-free milk amino acids were given by intravenous injection pump through the feeding tube. The amount of milk and fluids was gradually increased to normal amount according to the condition. In initial 3 days the intravenous nutrition was given and one week after operation, the infants were fed through mouth in addition to pumping milk through the tube and stopped infusion. Ten to 22 days after operation, the tube was removed and the infant patients were discharged., Result: All the five infants showed that the feeding through the nutrition tube was accomplished and the time of venous nutrition was reduced and fistula operation was avoided. None of the infants on question was off the tube and no jaundice exacerbation was found and the liver function was also found normal. At the very beginning, the tube was occasionally blocked by milk vale in one infant and after 0.9% sodium chloride solution flushing patency restored. After that, the feeding tube was washed once with warm water after feeding. In one infant vomiting occurred due to enough oral milk. The photograph of upper gastrointestine did not show anastomomotic stricture or fistula, or intestinal obstruction. After pulling out the tube, the symptoms disappeared and then the patient was discharged. One child was found to have diarrhea with no lactose nutrition liquid and given compound lactic bacteria preparations for oral administration, the symptom disappeared. In the 5 cases, the shortest hospital stay was 10 days and the longest was 22 days, the average stay was 16 days. Three to 5 days after operation the weight restored to birth weight, the weight had increased, when discharged, to an average of 5.5 g (kg·d)., Conclusion: The small intestinal feeding tube was very effective for the postoperative nutrition maintenance of high position intestinal obstruction in newborn infants.

Objective: To analyze the clinicopathological characteristics and prognosis of a rare histological type of esophageal cancer-sarcomatoid carcinoma., Methods: Clinicopathological data of 31 patients with esophageal sarcomatoid carcinoma who underwent surgery in the Department of Thoracic Surgery of Zhejiang Cancer Hospital from Jan 2000 to Dec 2009 were collected and analyzed. The survival analysis was performed using Kaplan-Meier method., Results: All the patients underwent surgery. Of the 31 patients, one received preoperative chemoradiotherapy and postoperative chemotherapy, and 8 received postoperative chemotherapy. All the tumors were located in the middle or lower esophagus. Microscopically, the tumors were composed of both carcinomatous and sarcomatous components, and there was a transition between the two components, but no obvious heterogenous elements such as osteosarcoma, chondrosarcoma or rhabdomyosarcoma were found. In the carcinomatous components, positive expression of CK and EMA was found in all the 31 cases, and positive expression of vimentin in 5 of the 31 cases. In the sarcomatous components, positive expression of CK, EMA and vimentin was found in 29, 28 and 23 cases, respectively. The 1-, 3-, and 5-year survival rates were 80.6%, 55.9% and 33.4%, respectively, and the median survival time was 40 months., Conclusions: Esophageal sarcomatoid carcinoma is a particular type of esophageal malignancy with unique clinicopathological features. The diversity and complexity of the carcinomatous and sarcomatous components and their potential of transformation and differentiation lead to different prognosis from each other.

Objective: To introduce a self-developed bone collector designed by the authors and evaluate its effect in clinical application., Methods: A self-developed bone collector was used in 38 patients who underwent the dental implant operation from May 2008 to October 2009. During the operation the bone particles were harvested, which was used for the reconstruction of the bone defects around the implant simultaneously. The defects were fenestrate in 29 regions and triangle in 23 regions on buccal or lingual side of the dental implants., Results: All cases had no surgical wound infection and dehiscence 7 - 10 days after operation. Three to six months after implantation, the X-ray showed that all the dental implants had perfect osseointergration and the alveolar bone were successfully augmented., Conclusions: Bone debris harvested by self-developed bone collector could repair small peri-implant bone defects, which is simple to use in clinic.

Objective: To explore the expressions and the significance of CD(3)(+)CD(16)(+)CD(56)(+) NKT cells, CD(3)(-)CD(16)(+)CD(56)(+) NK cells and T lymphocyte subsets in peripheral blood of patients with multi-drug resistant (MDR-TB) and extensively drug-resistant (XDR-TB) pulmonary tuberculosis., Methods: The data of 316 patients with pulmonary tuberculosis hospitalized in Shanghai Pulmonary Hospital from January 2008 to June 2009 were retrospectively analyzed, of whom 119 were newly diagnosed, and 197 were retreated patients. There were 204 males and 112 females, aged from 17 - 88 years, mean (44 ± 16) years. According to the results of drug-resistance, these patients were divided into a MDR group, an XDR group and a sensitive group. There were 146 patients in the MDR group, with 102 males and 44 females, aged from 19 - 84 years, mean (42 ± 16) years. There were 77 patients in the XDR group, with 42 males and 35 females, aged from 18 - 88 years, mean (50 ± 16) years. There were 93 patients in the susceptible group, with 60 males and 33 females, aged from 17 - 83 years, mean (43 ± 19) years. According to the distribution of cavitation in lung fields, these patients were also divided into 1 - 2 lung field affected group (n = 132), 3 - 4 lung field affected group (n = 49) and 5 - 6 lung field affected group (n = 9). The frequencies of NKT cells, NK cells and T cells from whole blood were tested by flow cytometry. Rank test (SAS software) was used for statistic analyses., Results: The expression rate of NKT cells and NK cells was the highest in the XDR group [11% (6% - 16%) and 7% (4% - 12%)], as compared to the MDR group [8% (5% - 14%) and 6% (4% - 11%)], and the susceptible group [7% (4% - 11%) and 5% (3% - 9%)], the difference being statistically significant (H = 6.478 - 8.369, P < 0.05). The expression rate of the NKT (14 ± 9)% and NK cells (11 ± 7)% in males of the XDR group was significantly higher than that in females [NKT (9 ± 5)% and NK cell (6 ± 4)%], while CD(4) (38 ± 10)% and CD(4)/CD(8) (1.9 ± 1.3) were significantly lower than those of the females [CD(4) (44 ± 10)% and CD(4)/CD(8)(2.2 ± 0.7)], the difference being statistically significant (z = -2.91 - -2.79, P < 0.05, P < 0.01). The expression rate of CD(4) (42 ± 9)% was the highest, but CD(8) (22 ± 8)% was the lowest in the 1 - 2 lung field group. While in the 5 - 6 lung field group, the expression rate of CD(4) (36 ± 11)% was the lowest, CD(8) (28 ± 12)% was the highest, and CD(4)/CD(8) (1.5 ± 0.8) was the lowest, the difference being statistically significant (H = 8.404 - 16.175, P < 0.01)., Conclusions: With the increasing level of drug resistance, the expression rate of NKT cells and NK cells increased, while the expression of T cell subsets did not change. The value of CD(4) and CD(4)/CD(8) in peripheral blood decreased, but CD(8) increased as the extent of cavitation increased in these patients. The impairment of cellular immune function in XDR-TB was more prominent in male patients.

Objective: To investigate the differentiation potential of human bone marrow mesenchymal stem cells (hMSC) inducing into epithelial-like cells, even corneal epithelial-like cells, and to discuss the plasticity that make hMSC the seed cells used in corneal tissue engineering., Methods: hMSC were isolated and purified by density gradient centrifugation combined with an attachment culture method and passaged in vitro. hMSC were identified by flow cytometry. The passaged hMSC were planted on fresh pig corneal Bowman's membrane. The expression of CK12, ABCG2 and CK19 in hMSC was identified by immunofluorescence staining. We used in vitro method to obtain a multilayer culture of hMSC. When hMSC formed a monolayer, the cells were inserted to Millicell culture and grew into multilayers by using the air-lifting cultivation methodology. Four weeks later, after fixed and dehydrated, the hMSC were observed under the light microscope after hemotoxylin and eosin (HE) and immunohistochemistry staining., Results: hMSC could be cultured, expanded in vitro, and showed great potential of proliferation. The result of flow cytometry showed that the positive staining percentage was 0.06% for CD45, 0.41% for CD34, 86.43% for CD44, 85.72% for CD29 and 90.72% for CD105. This indicated that hMSC expressed CD44, CD29, CD105 but not CD45 and CD34. After four weeks induction, part of hMSC expressed CK12 and CK19 but not ABCG2. In the in vitro stratification, HE and immunohistochemical staining showed that there were one or two layers epithelial-like cells, even corneal epithelial-like cells after using the air-lifting cultivation., Conclusions: This study suggests that hMSC have the potential to differentiate into epithelial cells, even corneal epithelial cells. hMSC could be the option of cells used to reconstruct the corneal epithelium by tissue engineering technology.

Objective: To summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment., Method: The process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed., Results: The main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding., Conclusion: For children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.

Objective: To study the clinicopathological and immunohistochemical features, histogenesis and biological behavior, clinical treatment and prognosis of solid pseudopapillary tumor of the pancreas (SPT)., Methods: Routine HE and immunohistochemical (SP) stainings were used in the pathological examination of 18 cases of SPT. Their clinical data were retrospectively analyzed. All the 18 postoperative patients were followed-up for 3 months to 10 years with an average of 29.2 months., Results: There were 16 females and 2 males, age ranging from 9 to 65 years with mean age of 25.3 years. Abdominal pain and palpable mass were among the major complains. Tumors were encapsulated and mixed with solid and cystic tissues. Histological features were pseudopapillary structure with a fibrovascular core. Immunhistologically, most tumors were positive for alpha-AT, alpha-ACT and Vim, with a high percentage of 94.4%. The eighteen cases were followed-up from 3 to 120 months. Five cases received reoperation after recurrence, and 14 cases were alive. Maximum survival time was 121 months and the minimum survival time was 3 months, with a median survival time of 23.0 months. The 5-year survival rate was 72.2%. A Kaplan-Meier analysis revealed that patient's age, tumor size, pathologic features, metastasis were major prognostic factors for SPT., Conclusion: SPT is a tumor of low-grade malignancy and may be derived from multipotent stem cells. SPT most frequently affects young female, and has distinct clinicopathologic manifestation with excellent prognosis after surgical treatment.

Objective: To report nine cases of descending necrotizing mediastinitis (DNM) and to summarize the management experience., Methods: Between December 2005 and December 2008, nine patients (mean age, 55.7 years; age range, 38 to 78 years) with DNM were treated. Eight patients underwent surgical drainage of the involved cervical region and mediastinum (4 with cervical drainage alone; 4 with cervical drainage and right thoracotomy)., Results: Two patients died, one of them refused surgical therapy and the other one died of multiorgan failure related to postoperative septic shock. Seven patients recovered. The mortality rate was 22%., Conclusions: Delayed diagnosis and inadequate drainage are the main causes of high mortality rate of DNM. Aggressive surgical drainage and debridement of the neck and mediastinum by a multidisciplinary team of surgeons are very important in the treatment of DNM.

Objective: To study the efficacy of anti-telomerase siRNA in hepatocellular carcinoma both in vitro and in vivo., Methods: Lentvirus vectors contained anti-telomerase siRNA were conducted with a high performance homologous recombination system, and then were transduced into human hepatocellular carcinoma HepG2 cells. The telomerase activity was detected by RT-PCR, HepG2 cell proliferation was determined by MTT assay, and apoptosis was detected by TUNEL assay. The in vivo experiment was carried out by inoculation of HepG2 cells into nude mice and the tumor growth was measured and analyzed., Results: The growth of transfected HepG2 cells was significantly inhibited and the inhibition rate was 57.5% at the 8th day after transfection. The telomerase activity was significantly suppressed in vitro. The growth of transfected human hepatocellular HepG2 tumor in the nude mice was also significantly inhibited., Conclusion: The results of this study demonstrate that the growth of hepatocellular carcinoma cells is effectively inhibited by transfection of anti-telomerase siRNA both in vitro and in vivo.

Objective: To establish anisotropic mandible model with dental implants and to investigate the effect of anisotropy material on stress and strain distribution of implant-bone interface., Methods: Three-dimensional finite element models of whole mandible with anisotropic and equivalent isotropic material were created by CT scanning and universal surgical integration system (USIS) software developed by the authors. Two ITI threaded implants were implanted in the posterior teeth area. The values of principal stress and principal strain on the bone around dental implants were calculated in two different finite element models with buccolingual load., Results: In the anisotropic mandible model, nearly all values of the principal stress and principal strain on cortical and cancellous bone increased compared with the equivalent isotropy model, 2.1%-74.1% for principal stress and 4.7%-57.3% for principal strain, but 10. 3%-71.4% for principal stress and 19.5%-63.4% for principal strain on cancellous bone., Conclusions: In the three-dimensional finite element analysis, anisotropic mandible model with dental implants has an apparent effect on the stress and strains of the implant-bone interface. Anisotropic mechanical properties of mandible should be emphasized in biomechanical study.

Objective: To study the mechanism of enhancement of the CTL activity in mice co-expressing of CD80, CD86 and CD137L genes., Methods: The mice were randomly divided into five groups, named A, B, C, D and E. The group A and B were control groups (CG). H22-BAL B/c HCC mouse model was established by subcutaneous injection with hepatocellular carcinoma cells of cell line H22-Wt (group A), H22-neo (group B), H22-CD80/CD86(+) (group C), H22-CD137L(+) (group D) and H22-CD80/CD86/CD137L(+) (group E), respectively. On the 14th, 35th, 56th and 84th day after the first inoculation of tumor cells, TUNEL staining and DNA ladder examination were used to detect apoptosis of splenic T lymphocytes in all groups at each post-inoculation time point. Electrophoretic mobility-shift assay (EMSA) method was used to detect the activity of nuclear factor kappaB (NF-kappaB) in splenic T lymphocytes in each group at each time point post-inoculation., Results: Apoptosis was found in a great number of T lymphocytes in CG on the 14th day, much more than that in group C and E. The number of apoptotic T cells in group C had a significant difference compared with that in the group E from 14th to 84th day (P = 0.003). DNA ladder analysis showed typical positive results in group C and E. The significant apoptosis fragments were found in group C on 21st, 35th and 84th days. NF-kappaB activity of T cells in groups C and E was remarkably higher than that of groups CG and D, with higher in group D than that of CG (P = 0.002), and with no significant difference between group C and E on 14th day. The activity in group E was stable and remarkably higher than that of group C on 56th and 84th days after the first inoculation., Conclusion: H22-CD80/CD86/CD137L(+) induces higher NF-kappaB activity of the host T cells by synergistic action of CD28 and CD137, which may be one of the mechanisms of enhancement of the host CTL activity induced by co-expression of CD80, CD86 and CD137L genes.

Objective: To evaluate and compare effects of compound galvano-ceramic bridges with Ni-Cr alloy ceramic bridges and gold alloy ceramic bridges in clinic., Methods: A total of 105 bridges were made for 103 patients. Of them, 35 were compound galvano-ceramic bridges, 35 Ni-Cr alloy ceramic bridges, and 35 gold alloy ceramic bridges respectively. The marginal fitness and fracture of ceramic bridges were checked and caries or second caries of all abutments were also examined in clinic at 6 years after cementation., Results: The marginal fitness of compound galvano-ceramic bridges was evaluated as good in 100% of the bridges, which was better than the fitness of gold alloy ceramic bridges (91%) and Ni-Cr alloy ceramic bridges (77%) (P < 0.01). There was no difference in fracture of ceramic layer among compound galvano-ceramic, gold alloy ceramic, and Ni-Cr alloy ceramic bridges at 6 years (P > 0.05). None of compound galvano-ceramic bridge was found out caries or second caries of abutments. Two of 35 (6%) gold alloy ceramic bridges was found out second caries of abutments and five of 35 (14%) Ni-Cr alloy ceramic bridges were found out caries or second caries of abutments at 6 years. None of frameworks of all bridges was loose or broken., Conclusions: The 6-year clinical study on compound galvano-ceramic bridges presents good results in marginal fitness and fracture resistance, which indicates the compound galvano-ceramic bridge can be used in clinic.

Objective: To introduce the internal tri-focal distractor developed by the authors and evaluate its primary application and feasibility in animal experiment., Methods: Four hybrid dogs were selected and segmental resection at the mandibular symphysis was performed. Two transfer disks, 10 mm in width, were respectively fixed at the two ends of bone defect and the internal tri-focal distractor was installed. The force was applied one week after the operation, with a tractive speed of 0.5 mm/12 h. After 13 - 16 days, the traction was completed when the two transfer disks were combined. The occlusal films were taken at the 1st day, 4th, 8th and 12th week after traction. The animals were killed at the 12th week, samples of the traction area taken and histological examination performed. Finally, the new bone formation was observed., Results: All the animals survived in the experiment, abruption and rust-eaten was not found in distractor. In the 12th week after installation, new bones in traction gap were mature and bony union between the two transfer dishes was accomplished., Conclusions: The internal tri-focal distractor used in this study may become a potential device in reconstruction of mandibular symphyseal defects.

Objective: To understand the influence of co-expression of CD80, CD86 and CD137L genes on tumor immunogenicity in hepatocellular carcinoma H22-BAL B/c mouse models., Methods: The mice were randomly divided into five groups, named A, B, C, D and E, and control groups A and B, 20 mice in each group. Hepatocellular carcinoma H22-BAL B/c mouse model was established by subcutaneous injection of cells H22-Wt, H22-neo, H22-CD80/CD86+, H22-CD137L+ and H22-CD80/CD86/CD137L+, respectively. The rate and incubation period of tumor development, survival rate, and the tumor growth in vivo were observed and recorded. The effects of gene transduction on immunogenicity of the tumor and antitumor immunity of the animals were assessed by re-innoculation of wild type H22 cells., Results: The rate of tumor development in group E was only 50%, much lower than that in other four groups (P < 0. 01). The tumor growth in group C was reduced with complete tumor regression in two hosts (20%, 2/10). In group E, there was more pronounced reduction of tumor size. The maximal tumor sizes were remarkably smaller than those of group C, and there was complete tumor regression in three mice (60%, 3/5). No tumor regression was found in the other three groups. Survival rates of group C and E were significantly higher than that of animals in groups A, B and D (P < 0. 01), but no significant difference was seen between group C and E. The results of re-inoculation test showed that tumor formation rate was 40% (4/10) in group C, 100% (8/8) in group D, and 0 (0/5) in group E. There were significant differences between groups C and E and control group, between group E and C, but not between C and D. After the third time of re-inoculation with H22-Wt cells at the 56th day, tumor occurred in 6/6 mice (100%) of group C, but 0 (0/5) in group E. The difference was very significant. Five animals without tumor formation after the first inoculation in group E, were re-inoculated with H22-Wt cells on the 21st day and the third re-inoculation on the 56th day, no tumor was found (0/5)., Conclusion: Both co-expression and solo-expression of CD80+ CD86 and CD137L genes reduce the tumorigenicity of wild type H22 cells, but co-expression of CD80, CD86 and CD137L genes can more significantly improve the immunogenicity of H22-CD80/CD86/CD137L+ cells.

Objective: Gene therapy of leukemia is a new and effective method. It is known to all that the pathogenesis and development of leukemia are related to a variety of genes. Survivin is a member of inhibitors of apoptotic proteins (IAP). Its cDNA was cloned from target cell protease receptor-1 (EPR-1). It is expressed in common tumors, but there is no expression in normal and mature tissues. High expression of survivin was detected in leukemic cells. The present study was conducted to examine the role of survivin in the differentiation of leukemic cells by using antisense-oligonucleotides., Methods: Human leukemic cell K562 was used as the model for the study. K562 cells were divided into 4 groups randomly: antisense oligonucleuotide (ASON) group, nonsense oligonucleotide (NSON) group, lipofectin group and control group. There were 5 samples in each group, and the experiment was repeated for three times. ASON was designed with the reference to targeting survivin mRNA. K562 cells were cultured in RPMI1640 contained fetal cattle serum at a concentration of about 10 percent. Cell transfection was induced by lipofectin. Forty-eight hours after thansfection, the morphology and ultrastructure were observed. Twenty-four hours and 48 hours after thansfection, the function of K562 cells was detected by benzidine staining, POX staining and NBT staining, respectively. The mean fluorescence intensity of CD33 was determined by flow cytometry. The method of immunohistochemistry was used to examine the protein level of survivin., Results: After thansfection with ASON, the size of K562 cells was reduced, but the cytoplasm was increased. The metarubricyte, segment granulocyte, apoptotic cells could be found. Morphologically and ultrastructurally, erythroid and myelocytic differentiation was observed. The positive level of benzidine staining in ASON group (11.90 +/- 2.30 at 24 h and 18.20 +/- 2.93 at 48 h) was higher than that of NSON group, lipofectin group and control group, respectively. The positive level of POX staining in ASON group (17.40 +/- 3.54 at 24 h and 29.40 +/- 3.70 at 48 h) was also higher than that of any other groups. The positive level of NBT staining in ASON group (7.50 +/- 2.26 at 24 h and 12.10 +/- 2.63 at 48 h) was significantly higher than that of NSON group, lipofectin group and control group, respectively (P < 0.01). In ASON group, the mean fluorescence intensity of CD33 (21.43 +/- 1.61 at 24 h and 14.86 +/- 1.20 at 48 h) was significantly lower than that of any other groups (P < 0.01). After thansfection for 24 h, the protein level of survivin in ASON group was decreased significantly compared to that of control group. There was no difference in survivin protein level amongst ASON group, NSON group and lipofectin group at 24 h (P > 0.05). Forty-eight hours after thansfection, the protein level of survivin was decreased significantly., Conclusions: ASON targeting survivin can induce K562 to erythroid and myelocytic differentiation. Survivin is related to differentiation of K562 cells, and it can be a target of gene therapy for leukemia.

Objective: To analyze the prognostic factors in patients with cervical squamous cell carcinoma of stage Ib and IIa treated by surgery, and to investigate their guide roles in available post-operation adjuvant therapy., Methods: The clinicopathologic records of 306 patients with cervical squamous cell carcinoma of stage Ib and IIa who underwent radical hysterectomy and pelvic lymphadenectomy were retrospectively analyzed, and the prognostic factors were explored by univariate and multivariate methods. Independent prognostic factors were identified by COX proportional hazards regression model., Results: The overall 5-year survival rate of these 306 patients was 78.1%. In univariate survival analysis, the poor prognostic factors included poor differentiation, positive pelvic lymph nodes, deep stromal invasion, parametrial extension, tumor size > or = 4 cm, and lymph vascular space involvement (P < 0.05). While in multivariate survival analysis, the independent prognostic factors included positive pelvic lymph nodes, deep stromal invasion and parametrial extension (P < 0.05). According to the risk factors, all patients were divided into low, intermediate and high risk groups with 5-year survival rate of 90.3%, 83.9% and 43.1%, respectively. In low risk group, no risk factor or only parametrial extension was involved, pelvic recurrence rate was only 2.2%. In intermediate risk group, deep stromal invasion or together with parametrial extension was involved. Pelvic recurrence rate and extra pelvic metastasis rate were 13.5% and 1.3%, respectively. In high risk group, lymph node metastasis or together with other high risk factors was involved, pelvic recurrence rate and extra pelvic metastasis rate were 25.9% and 48.3%, respectively, and 10.3% had both pelvic recurrence and extra pelvic metastasis., Conclusions: Lymph node metastasis, deep stromal invasion and parametrial extension were independent prognostic factors of stage Ib and IIa cervical squamous cell carcinoma patients undergoing radical hysterectomy and lymphadenectomy. The establishment of prognosis-predicting system based on high risk factors may be helpful to predict the risk of recurrence and metastasis, and guide adjuvant therapy after surgery.

Objective: The shear fracture strengths of carbon fiber post, IPS-Empress 2 all-ceramic post (without ZrO2 post), cast Ni-Cr alloy post, prefabricated zirconia ceramic post, human dentin and the shear bonding strengths of the first three kinds of post bonded in the human root canals were measured and compared, which are as the reference for dental clinic., Methods: Standard cylindrical samples were made respectively in each group of carbon fiber post, IPS-Empress 2 all-ceramic post, cast Ni-Cr alloy post, prefabricated zirconia ceramic post, human dentin, three for each group. The shear fracture strengths of these samples were measured by universal testing machine (AUTOGRAPH DCS5000). Standard cylindrical samples were made respectively in each group of carbon fiber post, IPS-Empress 2 all-ceramic post (without ZrO2 post), cast Ni-Cr alloy post, five for each group. They were bonded in extracted human root canals that had been prepared to 3.0 mm length and 2.0 mm diameter with Glass ionomer cement (Japan Shofu). The shear bond strengths were measured by the same universal testing machine., Results: The shear fracture strengths of the carbon fiber post (199MPa), cast Ni-Cr alloy post (210MPa, shear bend strength) and prefabricated zirconia ceramic post (193MPa) were statistically higher than those of IPS-Empress 2 all-ceramic post (109MPa) and human dentin (100MPa). The shear fracture strength of the carbon fiber post was statistically similar to that of cast Ni-Cr alloy post and prefabricated zirconia ceramic post. There were no statistical differences between the shear bond strengths of carbon fiber post (2.4MPa) and cast Ni-Cr alloy post (3.8MPa). IPS-Empress 2 all-ceramic post broke before debonding (2.7, break value)., Conclusions: Carbon fiber post, as well as cast Ni-Cr alloy post and prefabricated zirconia ceramic post, has a comparatively high shear fracture strength. The shear bond strengths of carbon fiber post is similar to cast Ni-Cr alloy post.

Objective: To study the construction of the PCI-neo mammalian expression vector system containing murine 4-1BBL gene and its stable and effective expression in rat hepatocellular carcinoma cell line CBRH7919., Methods: The murine full-length 4-1BBL cDNA was obtained and subcloned into the PCI-neo mammalian expression vector. The recombinant named as PCI-neo-4-1BBL was identificated by restriction enzyme digestion and sequencing. Subsequently, PCI-neo-4-1BBL was transfected into CBRH7919 with lipofectamine reagent, then G418-resistance clone was acquired and named as PCI-neo-4-1BBL-CBRH7919+. The stable expression of 4-1BBL mRNA in PCI-neo-4-1BBL-CBRH7919+ was detected by RT-PCR., Results: The fragment of 980 bp (4-1BBL) and 5.4 kb (PCI-neo) was shown after PCI-neo-4-1BBL had been digested by EcoR I and Not I and agarose gel electrophoresis. The DNA sequencing of 4-1BBL, proved to be identical to the data of 4-1BBL in Genebank, showed stable and effective expression in PCI-neo-4-1BBL-CBRH7919+., Conclusion: The PCI-neo mammalian expression vector system containing murine 4-1BBL has been constructed successfully, which shows stable and effective expression in CBRH7919.