OBJECTIVE: Animal experiments prove that alcohol may cause adipo-accumulation in femoral head and then induce osteonecrosis,but its mechanism of action is not known. NIH 3T3 fibroblast is a kind of pluripotent stem cell.There are not relative reports on the effect of alcohol on NIH 3T3fibroblast at present.OBJECTIVE: To observe the effect of alcohol on the differentiation of NIH 3T3 fibroblast into adipocyte and the influence of that on the expression of adipogenic transcription factor-peroxisome proliferator activated receptor gamma (PPAR-γ).DESIGN: Single sample observation.SETTING: Department of Biochemistry, Basic Medical College,Zhengzhou University and Department of Orthopaedics, First Affiliated Hospital, Zhengzhou University.MATERIALS: The experiment was completed in the Laboratory of Department of Biochemistry, Basic Medical College, Zhengzhou University from April 2002 to June 2003.NIH 3T3 fibroblasts were provided by the Laboratory of Department of Orthopaedics,University of Virginia Hospital.METHODS: The NIH 3T3 fibroblasts were divided into 6 groups randomly.The alcohols (volume fraction was 0.998) with the dosage 0(control group),0.03,0.06,0.09,0.15,0.21 mol/L were added respectively to the NIH 3T3 fibroblasts daily.The mixtures were cultured for 14 days and then stained with Sudan Ⅳ .The percentage of adipocyte was determined with image analysis software of computer,Image Pro Plus 4.1.The expression of PPAR-γ mRNA was determined with the technology of reverse transcription-polymerase chain reaction.alcohol and control groups.RESULTS: The cells were processed with the alcohols of progressive inpercentages of adipocytes in all the alcohol groups were 11.9% ,23.8%,28.2% ,31.1% ,42.6% respectively and 1.2,2.5,2.9,3.2,4.4 times that in control group(9.6%) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant (P ＞ 0.05),but the differences between the other 4 groups and control group,0.03 mol/L alcohol ues of PPAR-γ mRNA to 18S gene in the cells of all the groups were 0.218,0.411,0.486,0.473,0.453 respectively and were 1.1,2.1,2.5,2.4,2.3times that in control group (0.197) respectively. The differences between 0.03 mol/L alcohol group and control group were not significant(P ＞ 0.05),but the differences between the other 4 groups and control group and 003 mol/L alcohol group were significant extremely(P ＜ 0.001).CONCLUSION: Alcohol can induce differentiations of a large quantity of NIH 3T3 fibroblasts into adipocytes directly, which might be one of the factors of lipotrophy in bone marrow wheri alcohol-induced osteonecrosis happens.

BACKGROUND: Gene transfer to study the effect of a exogenous gene on cells is a commonly used technique for molecular biology. Compared with viral vector,pcDNA4.0-TGF-?1 eukaryotic expression plasmid transfection has no inherent toxicity or cytotoxicity,with high safety. OBJECTIVE: To construct and identify eukaryotic expression plasmid carrying recombined TGF-?1 by cloning transforming growth factor (TGF)-?1 gene. DESIGN,TIME AND SETTING: The in vitro cell experiment was performed at the Opening Laboratory of Key Clinical Medical Sciences,Henan Provincial High-learning School from March 2007 to February 2008. MATERIALS: Clean healthy Sprague Dawley rats aged 2 months,of both gender,were used for isolating RNA from cells. METHODS: The TGF-?1 mRNA was extracted from the rat bone marrow cells,and amplified by reverse transcription-polymerase chain reaction (RT-PCR),and cloned into the pGEM-T-vector and identified by PCR. Then TGF-?1 gene was subcloned into pcDNA4.0 plasmid vector. The inserting DNA sequences were analyzed. MAIN OUTCOME MEASURES: Expression of TGF-?1 cDNA,pGEM-T-TGF-?1 and recon pcDNA4.0-TGF-?1. RESULTS: There was an obvious electrophoresis strip of TGF-?1 cDNA by RT-PCR. The obtained pGEM-T-TGF-?1 was confirmed correct with PCR amplification and sequence identification. Recombinant eukaryotic expression plasmid pcDNA4.0-TGF-?1 was obtained and identified by digestion. The inserted sequences were conformity to the designed sequences. CONCLUSION: Rat TGF-?1 gene has been successfully cloned,and recombinant TGF-?1 eukaryotic expression vector has been constructed successfully.

Objective To observe the effect of methylprednisolone (MP) pre-intervention on ex-pressions of heat shock protein 27 (HSP27) and tumor necrosis factor alpha (TNF-α) in cells in rat spinal cord following ischemia-reperfusion injury. Methods One hundred and fifty male Sprague-Dawley (SD) rats were randomly divided into 3 equal groups: group A (control) in which the abdominal aorta was exposed without any treatment, group B in which the abdominal aorta was clipped for 30 minutes before reperfusion for 3 bours to establish a model of ischemia- reperfusion injury, and group C in which intravenous MP injection was conducted 30 minutes before the establishment of the ischemia-reperfusion injury model. Three hours later the spinal cords were harvested. Pathological changes of spinal cord cells were observed with HE staining and expressions of HSP27 and TNF-α in spinal cord cells were observed with immunohistochemical staining. The motor function of hind-limbs before was evaluated before sample harvest. The data were analyzed with SPSS software. Results There were significant differences between groups A and B in the expressions of TNF-α and HSP27. Compared with group B, the expression of TNF-α decreased and HSP27 increased in group C, with statistically significant differences between the 2 groups. The motor function score of hind-limbs decreased in group B but improved in group C. Conclusions Since MP can decrease the expression of TNF-α and up-regulate the expression of HSP27, it has a potency of neuro-protection. Spinal cord ischemia-reperfusion injury can be avoided or decreased after MP pre-intervention.

Objective To explore the surgical method, clinical result, biomechanical property, and surgical indication of treatment of thoracolumbar burst fractures with titanium mesh fusion device. Methods 22 patients with the thoracolumbar burst fracture and paralysis were treated by anterior decompression and spinal stability reconstruction with titanium mesh fusion device and a titanium plate. In this group, the fracture was at the 12th thoracic vertebra in 7 cases, the 1st lumbar in 14, and the 2nd lumbar in 1. According to Frankel’s classification, 4 cases were of Grade A, 6 of Grade B, and 12 of Grade C. 20 patients experienced the kyphosis deformity. The kyphosis angles varied from 12 to 29 degrees with a mean of 19.5 degrees. Results The period of postoperative follow up varied from 0.6 to 2 years with an average of 1.2 years. The neural function was improved for more than 1 grade. 15 cases could control urination after the surgery. The kyphosis angles were reduced to 3 to 8 degrees with a mean of 5 degrees. The complications caused by the implants were not found. Conclusions To treat the thoracolumbar burst fractures with paralysis, the compressive elements from anterior side of the spinal cord should be completely removed. Since the titanium mesh fusion device plus a plate can reconstruct the spinal stability that satisfies spinal biomechanical property, it presents important application value.

BACKGROUND: Animal tests verified that alcohol could cause lipid accumulation in femoral head, leading to osteonecrosis. Molecular biology tests verified that alcohol could induce differentiation of marrow stromal cells (MSCs) into adipocytes. If a drug can resist the differentiation of MSCs into adipocytes induced by alcohol, it can prevent and treat alcoholinduced osteonecrosis.OBJECTIVE: To observe the effects of inhibition of puerarin on differentiation of MSCs into adipocytes induced by alcohol from the aspect of cell biology.DESIGN: Randomized controlled experiment.SETTING: Department of Orthopaedics, First Affiliated Hospital,Zhengzhou University and Department of Biochemistry and Molecular Biology, Basic Medical College, Zhengzhou University.MATERIALS: The experiment was conducted at the Department of Biochemistry and Molecular Biology, Basic Medical College, Zhengzhou University from April 2000 to December 2002. A total of 50 clean healthy Kunming mice aged 6-8 weeks, of either sex, were selected. Bone marrow cells of the bilateral femur were obtained, with the inoculated density of 1.5×106/cm2, implanted into 6 wells and 24 wells cultured plate, randomized into groups, a well as a specimen and 24 specimens in each group.METHODS: MSCs were separated, cultured and assigned into 3 groups at random: model group (cells were treated with 0.09 mol/L alcohol), experimental group (0.09 mol/L alcohol and 0.01 mg/mL final concentration of puerarin) and controlled group (no alcohol and puerarin). Sudan Ⅲ staining was performed and adipose cells was counted under light microscope.Content of triacylglycerol, activity of alkaline phosphatase and content of osteocalcin in cell culture fluid were measured.MAIN OUTCOME MEASURES: ①Result of differentiation of MSCs into adipocytes in micé of the three groups, ②content of triacylglycerol in MSCs of mice of the three groups, ③activity of alkaline phosphatase in MSCs of mice of the three groups, ④content of osteocalcin in cell culture fluid in the three groups.RESULTS: ①After being disposed and cultured for 21 days, number of adipocytes in the experimental group and control group decreased markedly compared with the model group. The number of adipocytes in the model group was 8.9 times of that in the experimental group, and 15.6 times of that in the control group [(319.17±19.92), (35.92±23.77)(20.42±12.15)per cm2,P＜0.001]. ②After being disposed and cultured for 21 days, coutent of triacylglycerol in the experinental group and control group was remarkably lower than that in the model group [(4.15±1.92) and (3.42±1.60),(11.55±4.42) μgin a well, P ＜ 0.001]. ③After being disposed and cultured for 12 days, activity of alkaline phosphatase in the experimental group and control group was distinctly higher than that in the model group [(9.51±2.96),(11.18±3.11), (4.84±2.23) μkat/g,P ＜ 0.001]. ④After being disposed and cultured for 14 days, content of osteocalcin in the experimental group and control group was 2.2 times and 2.7 times of that in the model group, respectively, and the difference was very significant com pared with themodel group [(11.11±4.57),(13.43±5.29), (4.95±2.31) μg/g,P ＜ 0.001].CONCLUSION: Puerarin can inhibit the differentiation of MSCs into adipocytes induced by alcohol, and maintain the osteogenic differentiation,which may prevent the development of osteonecrosis.

ObjectiveTo explore the clinical diagnosis and surgical treatment of primary retroperitonealneurilemoma(schwannoma). MethodsClinicaldataof 47patientsof primary retroperitoneal schwannoma admitted and surgically treated from January 1995 to December 2009 were retrospectivelly reviewed.ResultsAs diagnosed by pathology there were 36 cases of Benign schwannoma,with a median age at onset of 41years, among those 11 patients were symptomatic, and 25 were asymptomatic. There were 11 malignant 11 cases, the median age was 38 years, among those 6 patients were symptomatic, and 5 were asymptomatic. The positive diagnostic rate of preoperative CT and MRI were 36. 2％ ( 17/47 ) and 58. 3％ ( 7/12 ) respectively. Immunohistochemically positive rates of S-100 were 100％ and 81.8％(9/11) in benign and malignant group respectively.All cases underwent surgical treatment. Surgical resection rates for benign and malignant groups were 100％ and 90. 9％(10/11)respectively. There was no perioperative death, Overall 5-year survival rates were 100％ and 45.5％ for benign and malignant tumors groups respectively. In benign group 2 cases recurred, in malignant group 4 cases recurred, and 3 had distant metastasis.ConclusionsPrimary retroperitoneal schwannomas are less common. It is difficult to make an accurate preoperative diagnosis. Surgery is the most effective therapy.Prognosis is good for benign and poor for malignant retroperitoneal neurilemomas.

BACKGROUND:Deep vein thrombosis and pulmonary embolism are certainly related to seasons.
OBJECTIVE:To analyze whether there is a seasonal pattern for deep venous thrombosis (DVT) after total hip and knee arthroplasty.
METHODS:We retrospected 866 patients (1 114 joints) underwent total hip and knee arthroplasty. There were 506 cases of total hip arthroplasty, including 287 male cases and 219 female cases, and 608 cases of total knee arthroplasty, including 133 male cases and 475 female cases. The average age was (60.98±0.87) years. After arthroplasty, patients were given color Doppler ultrasound in the deep veins of the lower limbs and pelvis to analyze the correlation between occurrence of deep vein thrombosis and seasons.
RESULTS AND CONCLUSION:There was a significant difference in the incidence of deep vein thrombosis in summer and winter (χ2=7.190, P=0.007), in summer and spring (χ2=6.995, P=0.008), as wel as in summer and autumn (χ2=5.663, P=0.017). No statistical differences were tested in spring and autumn, in autumn and winter, and in spring and winter (P>0.05). These findings indicate that there is a seasonal variation of deep venous thrombosis after arthroplasty, and the highest incidence of deep vein thrombosis is in winter.

Objective To evaluate the comprehensive medical goal appraisal system on hand hygiene compliance rate of health care workers(HCWs).Methods Comprehensive medical goal appraisal system was adopted to inter-vene hand hygiene compliance rate of HCWs in a comprehensive hospital ,hand hygiene compliance rates of HCWs and consumption of instant hand sanitizer per bed-day before (December 2012)and after intervention (January 2013-June 2014)were compared.Results Hand hygiene compliance rate after intervention was higher than before interven-tion (85.17% [18 208/21 379]vs 39.92%[853/2 137]),hand hygiene compliance rate enhanced by 113.35%(χ2 =2 590. 81,P <0.001).Hand hygiene compliance rates of HCWs of different departments,different occupations and different hand hygiene moments were all higher than before intervention (all P <0.001);after intervention ,hand hygiene compliance rate revealed a increased tendency,and has maintained high since October 2013 (>90%),consumption of instant hand sanitizer before and after intervention was 7.24 mL/bed-day(4 200 L/579 841 bed-day)and 10.54 mL/bed-day(9 323.5L/884 489 bed-day)respectively,the consumption after intervention increased by 45.58% compared with that before intervention. Conclusion Comprehensive medical goal appraisal can effectively enhance hand hygiene compliance rate ,and maintains at a high level;the measure can affect hand hygiene behavior of HCWs by hawthorne effect,and is an effective and long-term measure to improve hand hygiene compliance of HCWs.

BACKGROUND:Anterior subtotal corpectomy,decompression and fusion is a conventional method to treat cervical degenerative diseases.A titanium cage is an important implant to maintain the stability of the cervical spine after subtotal corpectomy.In recent years,many patients have complications such as titanium cage sinking,which are highly controversial. OBJECTIVE:To investigate the internal biomechanical relationship between the tilt angle of the titanium cage and postoperative titanium cage subsidence after anterior subtotal cervical corpectomy,decompression and fusion. METHODS:A three-dimensional finite element model of the C4-C6 segment was established by CT images of a normal human cervical spine,in which the anterior subtotal resection,decompression and fusion of the C5 vertebral body were simulated,and titanium cages with different tilt angles(-6° to-1° negative angle,that is,the front edge of titanium cage is shorter than the rear edge of titanium cage;1° to 6° positive angle,that is,the front edge of titanium cage is longer than the rear edge of titanium cage)were placed.After setting the boundary conditions,preloads of 50,100 and 150 N were applied respectively on the C4 vertebral body.The stress value of each contact point between the titanium cage and C4 lower-end plate and C6 lower-end plate(seven stress contact points on the contact surface of titanium mesh)was recorded and statistical analysis was conducted. RESULTS AND CONCLUSION:(1)The tilt angles of the titanium cage of the positive angle group and negative angle group under 50,100 and 150 N stress respectively were found by Mann Whitn test,with P<0.05,which was statistically significant.The dispersion coefficients of the positive angle group were smaller than those of the negative angle group under 50,100 and 150 N stress conditions.(2)Under 50,100 and 150 N stress conditions,the Wilcoxon sign rank test in the positive angle group of titanium cage tilt angle found that when the angle was set to 1° to 5°,the difference was not statistically significant(P>0.05).However,when the tilt angle of the titanium cage was set to 6°,the difference was statistically significant(P<0.05).(3)Under 50,100 and 150 N stress conditions,the Wilcoxon sign rank test in the negative angle group of titanium cage tilt angle found that when the tilt angle was set to-1° to-6°,the difference was not statistically significant(P>0.05).(4)It is concluded that in the sagittal position,the titanium cage with a positive tilt angle is more stable than with a negative tilt angle,which is more suitable for clinical use.The tilt angle of the titanium cage is relatively stable in the range of 1° to 5°.When the tilt angle is 6°,the stability starts to decline,which is easy to cause complications of titanium cage sinking after surgery.It is more suitable to select the titanium cage with a tilt angle of 1° to 5° according to the clinical situation during surgery to improve the efficacy.

Objective:To discuss the classification and treatment of ureteroileal anastomotic stricture (UAS) after radical cystectomy.Methods:The clinical data of 34 patients with UAS after radical cystectomy in the Department of Urology of Tongji Hospital from January 2017 to January 2022 were reviewed and analyzed. There were 25 males and 9 females. The average age was (66.3±7.7)years, including 2 cases of bilateral hydronephrosis and 32 cases of unilateral hydronephrosis. The average time of UAS was detected (14.7±6.5)months after radical cystectomy. There were 32 patients of unilateral hydronephrosis and 2 patients of bilateral hydronephrosis. Two patients had undergone nephrostomy in an external hospital. Three patients had elevated leukocytes in blood routine. Among them, two patients had fever. First, nephrostomy on the hydronephrosis side and anti-infection treatment were performed. After routine blood tests showed that the white blood cells were normal and antibiotics were stopped for 24 hours without fever, the operation was performed. 34 patients had preoperative hydronephrosis of (2.7±0.6) cm. Of the 34 cases in this group, 5 cases were injected with methylene blue through a preoperative nephrostomy tube, and 29 were injected with methylene blue through the renal pelvis using an 18G puncture needle under ultrasound guidance. Using a ureteroscope to observe in the ileal bladder, methylene blue was seen in 4 cases. Methylene blue was used to guide the search for the stenosis and a super smooth guide wire was inserted. Among them, 3 cases were dilated with a 5 mm ureteral dilation balloon catheter, 1 case was dilated with a F14 ureteral access sheath, and then a F6 single J stent was inserted. Methylene blue was not seen in the ileal conduit in 30 cases, of which 16 cases were treated with a flexible ureteroscope through the nephrostomy to locate the stenosis, incised with a 30 W holmium laser. 9 cases were treated with 5 mm ureteral dilation balloon catheter, and 7 cases were treated with a F14 ureteral access sheath, and then an F6 single J stent was inserted. 14 cases were unable to find the stenosis by antegrade method. According to the operation time and patient's condition, it was decided to perform immediate or second stage dual endoscope surgery. Through the nephrostomy, a flexible ureteroscope was used to enter the stenosis along the super slide guide wire. A rigid ureteroscope was used to observe the stenosis through the ileal conduit, and the stenosis was found. The stenosis was found in 10 cases and incised with a 30 W holmium laser. 8 cases were treated with 5 mm ureteral dilation balloon catheter, and 2 cases were treated with a F14 ureteral access sheath, and then an F6 single J stent was inserted. 4 cases were still unable to accurately locate the stenosis using the dual endoscope surgery(one case was bilateral stenosis, and one side was relieved), and continued indwelling nephrostomy. The definition of successful removal of stricture in this study is that an F6 single J stent can be inserted into the ureter.Results:UAS were classified into four types based on the severity of the intraoperative findings: Type Ⅰ, the narrow ureteral lumen is more than 50% narrower than the normal ureteral lumen, but methylene blue can pass through in strands; Type Ⅱ, needle like stricture of the ureteral lumen, allowing only methylene blue filaments to pass through; Type Ⅲ, membranous atresia of the ureter, with a narrow segment of 1 to 3 mm in length, and methylene blue cannot pass through; Type Ⅳ, long segment stenosis. Of the 34 cases in this group, 4 cases were type Ⅰ, and the stenosis was dredged by retrograde method; 16 cases were type Ⅱ, and the stenotic segments were dredged by antegrade method; 10 cases were type Ⅲ, and the stenosis was dredged by the dual endoscope surgery; Four cases were of type Ⅳ (one case was of bilateral UAS, one side was of type Ⅲ, and the other side was of type Ⅳ, which was classified as type Ⅳ). The stenotic segment could not be solved through the above methods. Among the 34 patients, 30 patients were successfully relieved of anastomotic obstruction, and 1 patient with bilateral obstruction was unilaterally relieved of anastomotic obstruction. In the other 3 cases, because the stenosis segment was too long, 2 cases were changed to nephrostomy, and 1 case was changed to open surgery, with a success rate of 88.2%. UAS was classified into 4 types based on the severity of UAS seen during surgery. No serious complications occurred during and after the operation. During the follow-up of 6-24 months, the imaging evaluation of 4 patients showed that hydronephrosis was aggravated, with an average increase in creatinine of (32.5±10.9)μmol/L, requiring replacement of a single J tube. The imaging evaluation of the remaining 26 patients showed that the postoperative hydronephrosis was 0.9 ± 0.6 cm less than the preoperative hydronephrosis 2.6 ± 0.6 cm, with a statistically significant difference ( P<0.01). The quality of life score at 3 months after surgery was (1.9±0.6), which was significantly improved compared to the preoperative indwelling nephrostomy period (5.2±0.7), with a statistically significant difference ( P<0.01) Conclusions:The treatment of UAS after radical cystectomy with retrograde, antegrade, and dual endoscope surgery has a high success rate, which can help some patients avoid the inconvenience of indwelling external drainage tubes and the risk of open surgery. Choosing an appropriate surgical method can achieve the goal of treating UAS with minimal trauma.