A Human Case of Lumbosacral Canal Sparganosis in China

In this study, we intended to describe a human case of lumbosacral canal sparganosis in People’s Republic of China (China). A 56-year-old man was admitted to Xiangya Hospital Central South University in Changsha, Hunan province, China after having an experience of perianal pain for a week. An enhancing mass, a tumor clinically suggested, was showed at the S1-S2 level of the lumbosacral spine by the examination of magnetic resonance imaging (MRI) with gadolinium contrast. The patient was received the laminectomy from S1 to S2, and an ivory-white living worm was detected in inferior margin of L5. In ELISA-test with cerebrospinal fluid (CSF) and serum samples, anti-sparganum antibodies were detected. He had a ingesting history of undercooked frog meat in his youth. By the present study, a human case of spinal sparganosis invaded in lumbosacral canal at the S1-S2 level was diagnosed in China. Although the surgical removal of larvae is known to be the best way of treatment for sparganosis, we administered the high-dosage of praziquantel, albendazole and dexamethasone to prevent the occurrence of another remain worms in this study.

Design of an axial blood pump of diffuser with splitter blades and cantilevered main blades / 生物医学工程学杂志

An implantable axial blood pump was designed according to the circulation assist requirement of severe heart failure patients of China. The design point was chosen at 3 L/min flow rate with 100 mm Hg pressure rise when the blood pump can provide flow rates of 2-7 L/min. The blood pump with good hemolytic and anti-thrombogenic property at widely operating range was designed by developing a structure that including the spindly rotor impeller structure and the diffuser with splitter blades and cantilevered main blades. Numerical simulation and particle image velocimetry (PIV) experiment were conducted to analyze the hydraulic, flow fields and hemolytic performance of the blood pump. The results showed that the blood pump could provide flow rates of 2-7 L/min with pressure rise of 60.0-151.3 mm Hg when the blood pump rotating from 7 000 to 11 000 r/min. After adding the splitter blades, the separation flow at the suction surface of the diffuser has been reduced efficiently. The cantilever structure changed the blade gap from shroud to hub that reduced the tangential velocity from 6.2 m/s to 4.3-1.1 m/s in blade gap. Moreover, the maximum scalar shear stress of the blood pump was 897.3 Pa, and the averaged scalar shear stress was 37.7 Pa. The hemolysis index of the blood pump was 0.168% calculated with Heuser's hemolysis model. The PIV and simulated results showed the overall agreement of flow field distribution in diffuser region. The blood damage caused by higher shear stress would be reduced by adopting the spindle rotor impeller and diffuser with splitter blades and cantilevered main blades. The blood could flow smoothly through the axial blood pump with satisfactory hydraulics performance and without separation flow.

Sulfated modification and DPPH free radical scavenging ability of polysaccharide from Rhodiola sachalinensis / 中草药

Objective: To optimize the sulfated modification conditions of Rhodiola sachalinensis polysaccharide (RSP) and improve the anti-oxidant activity of RSP. Methods: RSP were sulfated by chlorosulfonic acid-pyridine method, and the optimal conditions for the sulfated modification were determined by single factor experiments. The physical and chemical properties of RSP and sulfated RSP (S-RSP) were analyzed by infrared spectroscopy (IR) and scanning electron microscopy (SEM). The relation between substituting degree (DS) of S-RSP and anti-oxidative activity of polysaccharide was investigated by testing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging ability of RSP and S-RSP. Results: When the volume ratio of chlorosulfonic acid to pyridine was 1∶4, the reaction time was 2 h and the reaction temperature was 60 ℃, the maximum sulfur content of S-RSP was 18.83% and the DS was the highest for 2.38. Moreover, the anti-oxidant activity of RSP was enhanced by the sulfated modification, and there was a certain positive proportional relationship between DS and DPPH free radical scavenging ability of S-RSP. Conclusion: The volume ratio of chlorosulfonic acid to pyridine affects the DS of S-RSP, and the sulfated modification can increase the anti-oxidant capacity of RSP by changing its polarity.

Progress on strategies to promote vascularization in bone tissue engineering / 中国骨伤

With the continuous development of bone tissue engineering, a variety of emerging bone graft materials provided various methods for repairing bone defects. Early and rapid accomplishment of revascularization of materials interior after implantation of bone transplantation materials is a difficulty faced to bone tissue engineering. Blood vessels ingrowth provides the requisite netritional support for the regeneration reconstruction of bone tissue, for this reason, vascularization plays a significant role in bone tissue engineering. However,there is not a golden standard strategy of vascularization at present. Scaffold materials, cells and growth factors still are three indispensable elements in tissue engineering, and are cardinal points of the promoting vascularization strategies. Multiple growth factors or multiple cells combined with scaffolds, which are hot spots, have obtained excellent vascularization. This review focused on the comprehensive strategies for promoting the successful vascularization of tissue engineered scaffolds.

Protection of the superior petrosal vein in microneurosurgery for acoustic neuroma / 中南大学学报(医学版)

Objective:To explore the clinical signiifcance of the protection of superior petrosal vein (SPV) in the microneurosurgery for acoustic neuroma. Methods:From January 2009 to July 2011, 149 cases of acoustic neuroma microsurgery were observed. hTe difference in hematoma in surgical area, cerebellar hematoma and cerebellar edema were compared between a SPV without protection group (SPVWP group, n=8) and a SPV protection group (SPVP group, n=141). Results:In the 149 patients with acoustic neuroma, the SPV was reserved in 141 patients. In the SPVWP group (8 patients), hematoma in the surgery area occurred in 4 patients, cerebellar edema in 5, and cerebellar hemorrhage in 3. In the SPVP group (141 patients), hematoma in the surgery area occurred in 40 patients, cerebellar edema in 56, and cerebellar hemorrhage in 12. hTere was signiifcant difference in the incidence of cerebellar hemorrhage (χ2=3.84, P=0.05), no signiifcant difference in the incidence of hematoma in the surgical area (χ2=0.646, respectively, P=0.422), and no significant difference in the incidence of cerebellar edema (χ2=0.611, P=0.434) between the SPVWP group and the SPVP group. Conclusion:In acoustic neuroma surgery, the SPV should be protected, which may reduce the risk of cerebellar hemorrhage.

Study on the risk management for medical devices in use / 中国医疗器械杂志

Risk management penetrate the entire process of medical device regulation, and it is also very necessary for medical devices in use. Based on the analyzing of the status of risk management for medical devices, this paper discusses the principal, participants and entry point of risk management for medical devices.

Expression of fibroblast activation protein in HBV related hepatocellular carcinoma / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze the gene expression level of fibroblast activation protein in HBV related hepatocellular carcinoma patients and discuss its clinical significance.</p><p><b>METHODS</b>FAP gene expression in 33 hepatocellular carcinoma patients cancer tissues, peficancerous tissues, distant relative normal liver tissues and 13 normal liver tissues were examined by reverse transcription PCR; and real-time fluorescent quantitative PCR (qRT-PCR) was used to quantify their expression.</p><p><b>RESULTS</b>FAP were expressed in all the tissues,the relative expression values in cancer tissues, peficancerous tissues and distant relative normal liver tissues were 5.14 +/- 6.69, 1.58 +/- 0.96, 1.63 +/- 0.94, respectively, the differences were statistically significant (F = 4.401, P < 0.05); and in TNM stage I, II, IIII, they were 2.89 +/- 3.35, 4.15 +/- 4.69, 10.09 +/- 9.51 respectively; in well-differentiated, differentiated and poorly differentiated hepatocellular carcinoma were 1.62 +/- 1.74, 3.84 +/- 3.79, 1.26 +/- 13.34 respectively. The differences were all statistically significant (P < 0.05).</p><p><b>CONCLUSION</b>FAP may play an important role in the occurrence and development of HBV related hepatocellular carcinoma.</p>

Sequencing and analyses of the adenovirus polymerase gene in fecal samples of captively bred Rhesus macaques / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In an attempt to study the moleculr characterization and epidemiology of simian adenoviruses in nonhuman primate (NHP) populations.</p><p><b>METHODS</b>We examined a colony of captively bred rhesus macaques (Macaca mulatta) in China for the presence of adenoviral DNA in stool samples. This was done by using the PCR method that targeted the adenovirus polymerase gene, and the PCR positive fragments were cloned for sequencing and phylogenetic analyses.</p><p><b>RESULTS</b>Among the 57 animals analyzed, fecal samples from 12 animals were positive for the presence of adenoviral DNA. The results suggested that the viral DNA clones were primarily segregated into two large groups: SAdV-6 (2 non-redundant sequences) and SAdV-7 (9 non-redundant sequences). In addition, there were three clones with more similarity to SAdV-1, SAdV-3 and HAdV-52 respectively.</p><p><b>CONCLUSION</b>Our data confirmed the prevalence of adenoviral DNA in the feces of NHPs and revealed the adenoviruses in the gastrointestinal tract of the study animals. heterogeneity and phylogenetics of the adenoviruses in the gastrointestinal tract of the study animals.</p>

Association of single nucleotide polymorphisms of PATZ1 gene with azoospermia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the relationship between the polymorphisms of single nucleotide polymorphisms(SNPs) in rs2240424, rs2057951, rs2240427 and rs714909 loci in the PATZ1 gene and azoospermia.</p><p><b>METHODS</b>The allele and genotype frequencies of the four SNPs were investigated in 180 patients with azoospermia and 190 normal men as controls by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. The allele and genotype frequencies of the four SNPs were investigated in 180 patients with azoospermia and 190 normal men as controls by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis.</p><p><b>RESULTS</b>The frequencies of allele C (35.0% vs. 27.6%, P=0.031) and individuals with allele C (TC+CC) (57.8% vs. 46.3%, P=0.027) at the rs2057951 locus in azoospermic patients were significantly higher than that in normal men. There was a significant difference in distribution of haplotypes of the four SNPs between the two groups (P=0.01). Hapoltypes ACAC (11.1% vs. 6.6%, P=0.029) and ACGC (11.2% vs. 5.2%, P=0.003) increased significantly in azoospermic patients compared with controls.</p><p><b>CONCLUSION</b>The allele C of rs2057951 locus and haplotypes ACAC and ACGC of the four SNPs in PTAZ1 gene increased the susceptibility to azoospermia, suggesting that PATZ1gene may be associated with azoospermia.</p>

The comprehension of preservation for petrosal vein in 147 operative cases with acoustic neuroma / 中华显微外科杂志

Objective To discuss the preservation and clinical significance of petrosal vein in microsurgical operation of acoustic neuroma. Methods 147 patients with acoustic neuroma were operated, with internal decompression of the tumor firstly then dissected the tumor with surrounding structures, the petrosal vein were protected well in 143 cases and failed to protect in 4 cases. Results No hemorrhagic infarction in cerebellar was observed in 143 cases with intact petrosal vein. One case occurred with extensive cerebellar edema, which has gait disturbance after 18 months follow-up. The other three cases occurred with vein infarction and hemorrhagic edema after petrosal vein damage. One was dead and the other two were recovered well after decompression of posterior cranial fossa. One has no significant neurological deficit after 33 months follow-up, while the other has difficulty in line walking after 12 months follow-up. Conclusion Petrosal vein should be well protected in the operation of acoustic neuroma, the decompression of posterior cranial fossa should be considered if petrosal vein failed to protect.

Surgical outcomes of pediatric symptomatic epilepsy and their influencing factors / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the surgical outcomes of pediatric symptomatic epilepsy and the influencing factors for postoperative outcomes.</p><p><b>METHODS</b>A cohort of 48 children with symptomatic epilepsy received surgical treatment from October 2004 to September 2008. The surgical outcomes were followed up.</p><p><b>RESULTS</b>A 27.3 months (range 12-51 months) follow-up was performed in 43 cases. Engel classification for evaluating postoperative epileptic outcomes showed that class I in 32 cases (74%), class II in 4 cases (9%), class III in 4 cases (9%) and class IV in 3 cases (7%). Preoperative seizure frequency is an independent predictor of postoperative epileptic outcomes (P<0.05).</p><p><b>CONCLUSIONS</b>Operative treatment can lead to a favorable result in children with symptomatic epilepsy. Preoperative seizure frequency is an independent influencing factor for postoperative outcomes.</p>

Cell cycle arrest at M phase induced by vinblastine in MOLT-4 cells / 中国实验血液学杂志

This study was purposed to investigate the biological effect of vinblastine (VLS), usually known as inductor of mitotic arrest, on MOLT-4 of ALL cells and to evaluate its significance. The cell arrest in M phase and/or cell apoptosis were induced by treatment of MOLT-4 cells with 0.05 microg/ml VLS for 0 - 12 hours; the DNA histogram was detected by flow cytometry; the morphological changes of cells were observed by confocal microscopy; the cell cycle distribution, cell apoptosis and morphological changes of cells before and after arrest were analyzed by using arrest increasing rate (AIR), arrest efficiency (AE), apoptosis rate (AR) and morphologic parameters respectively. The results indicated that the cell arrest did not accompanied by significant increase of apoptosis rate; the DNA histogram of cell arrest showed dynamic change of cell cycle in time-dependent manner; the arrest efficiency could be quantified. The cell arrest at M phase was accompanied by cell stack in S phase, the cell proliferation rate dropped after cell arrest occurred. The cells arrested at M phase possessed of characteristic morphologic features in cell mitosis. It is concluded that the vinblastine can solely induce arrest of MOLT-4 cells at M phase. This study provides experimental basis for further investigating the relation of cell cycle arrest to apoptosis, mechanism of checkpoint and development of new anticancer drugs.

The significance of antimitochondrial IgA and IgG in the diagnosis of primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the sensitivity and the specificity of Anti-M2-3E ELISA for the detection of IgG- and IgA-specific isotypes of antimitochondrial antibody (AMA), and to investigate the significance of antimitochondrial IgA and IgG in the diagnosis of primary biliary cirrhosis (PBC).</p><p><b>METHODS</b>Sera were collected from 107 PBC patients, 87 disease controls and 26 healthy controls, and the antimitochondrial antibodies (IgG and IgA) were detected using indirect immunofluorescence (IFL), Anti-PDC ELISA and Anti-M2-3E ELISA.</p><p><b>RESULTS</b>The AMA IgG positive rate in PBC patients was 90.6% detected by Anti-M2-3E ELISA, which is significantly higher than that (81.3%) detected by IFL(t = 4.32, P < 0.05) and that (72.9%) detected by Anti- PDC ELISA (t = 6.03, P < 0.05). The AMA IgA was positive in 59 of the 107 PBC patients, and 99 of the 107 patients were positive for AMA IgG or/and IgA. 9 of the 20 IFL-negative patients were positive for AMA IgG as indicated by Anti-M2-3E ELISA, 11 of the 20 IFL-negative patients were positive for AMA IgG or/and IgA as indicated Anti-M2-3E ELISA. Compared to patients negative for IgG AMA, patients positive for IgG AMA had more severe histopathology and higher levels of ALP, IgG, and IgM.</p><p><b>CONCLUSION</b>The IgG and IgA Anti- M2-3E ELISAs are more sensitive for the AMA detection than IFN and the Anti-PDC ELISA. The presence of AMA IgG is the characteristics of severe PBC.</p>

Discrimination of anticancer agent action loci at G(2) and M phases by flow cytometry and confocal microscopic imaging / 中国实验血液学杂志

This study was purposed to evaluate a method to discriminate the action loci of anticancer agents in G(2) and M phases of cell cycle. The meta-amsacrine (m-AMSA) and vinblastine (VBL), already known as G(2) and M phase arrest agent respectively, were used to induce the arrest of MOLT-4 cells at G(2) and M phases, the change of DNA content was detected by flow cytometry, the morphology of arrested cells was observed by confocal microscopy so as to find the arrest efficacy difference of 2 anticancer agents. As a result, the flow cytometric detection showed that the arrested MOLT-4 cells displayed the raise of peaks in G(2) and M phases, but flow cytometric detection alone can not discriminate the difference between them. The observation with confocal microscopy showed that the MOLT-4 cells arrested by m-AMSA displayed the morphologic features in G(2) phase, while the MOLT-4 cells arrested by VBL displayed the morphologic features in M phase. This observation with confocal microscopy is helpful to discriminate the difference between them. In conclusion, the combination of flow cytometry with confocal microscopy is one of the effective methods to discriminate the kind of G(2) or M phase arresting agent of anticancer drugs.

Influence of chemotherapy on expression of TRAIL in primary acute leukemic cells / 中国实验血液学杂志

In order to explore the expression of TRAIL in primary acute leukemic cells and the effect of chemotherapeutic drug on TRAIL expression in acute leukemic cells, the expression of TRAIL was assessed by flow cytometry on day 0, day 1, day 3 and day 5 in 16 patients with acute leukemia received chemotherapy. Meanwhile, the bone marrow mononuclear cells of acute leukemia patients were cultured in vitro with VP-16 and INFalpha-2a. Expression of TRAIL was analyszed by flow cytometry at 24, 48 and 72 hours after treatment. The results showed that the expression of TRAIL in the peripheral blood mononuclear cells was upregulated significantly from day 1 after chemotherapy (P < 0.05). In in vitro culture test, VP-16 upregulated the expression of TRAIL on acute leukemia bone marrow mononuclear cells (P < 0.05). Compared with VP-16 alone, the combination of VP-16 with IFNalpha-2a showed no synergic effects on the expression of TRAIL. It is concluded that the expression of TRAIL increases after chemotherapy in vivo and after treatment with VP-16 and IFN in vitro, which suggests that the apoptosis induced by TRAIL may play an important role in chemotherapy of leukemia.

Influence and significance of intermittent ventilation on lung cytokines during cardiopulmonary bypass / 中南大学学报(医学版)

OBJECTIVE@#To determine the influence and significance of intermittent ventilation on lung cytokines during cardiopulmonary bypass (CPB).@*METHODS@#Thirty-six patients with rheumatic heart disease (RHD) were divided into 2 groups randomly: Treatment group (n = 18, given intermittent ventilation once every 5 minutes during CPB; control group ( n = 18), no ventilation during CPB. A brochioalveolar lavage was performed at 2 hours after the CPB. The numbers of granulocytes and the contents of IL-1, IL-6, TNF-alpha, and LDH in the brochioalveolar lavage fluids were measured, and lung PaO2/FiO2 were measured preoperatively and at 1 hour, 4 hours, and 8 hours after the CPB termination.@*RESULTS@#The numbers of granulocytes and the contents of IL-1, IL-6, TNF-alpha, and LDH of the treatment group in the brochioalveolar lavage fluids were significantly lower than those of the control group (P < 0.05), and the lung PaO2/FiO2 of the treatment group at 1 hour, 4 hours, and 8 hours after the CPB termination significantly increased than those of the control group (P < 0.05). A significant decrease of lung PaO2/FiO2 occurred in both groups at 1 hour, 4 hours, and 8 hours after the CPB compared with the same group at the baseline before the CPB (P < 0.01).@*CONCLUSION@#Intermittent ventilation has protective effects on the lung injury during CPB by decreasing granulocyte adhesion and the level of lung cytokines, alleviating the lung inflammatory reaction and endothelial cell injury.

The change of HBV nucleotide sequence during Adefovir dipivoxil treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the change of HBV nucleotide sequence during Adefovir dipivoxil treatment.</p><p><b>METHODS</b>Serum samples were collected from 4 patients (numbers 228, 229, 230 and 233) with chronic hepatitis B (CHB) treated with PMEA. PCR amplification of full-length genomes, total genome clone, and sequencing and linkage of the HBV viruses were done with the serum samples collected before the therapy, and again on the 28th week, 52nd week and on the 92nd week of the therapy. Gene types and serum types were studied according to the HBV DNA sequence. The related biological information was analyzed, including the homogeneity and heterogeneity of the HBV DNA sequence before and during PMEA therapy, and variance of amino acids coded by HBV sequence.</p><p><b>RESULTS</b>The gene types and serum types of HBV of the 4 patients were determined. Patient 228 was gene type C/serum type adrq+. Patient 229 was complex gene type B and C/complex serum type adw2 and adrq+. Patients 230 and 233 were both serum type adw2. The homogeneity of total gene sequence was 97.5% - 99.8% in one group of patients and 90.6% - 100% between groups in variant patients. The base mutation quantity of 52W was significantly higher than that of 28W and 92W. There was a common site of hotspot congregating mutation that existed in the P region. The target of PMEA was located in the P region encoding reverse transcriptase, which was generally very conservative. Therefore the mutations in this region were very significant. This might affect the activity of reverse transcriptase, leading to resistance to PMEA.</p><p><b>CONCLUSION</b>We discovered some unusual aminophenol variations in the HBV genomes, which most probably are related to the HBV mutation that resulted in the developing resistance to PMEA.</p>

Combined surgical therapy for hepatolithiasis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the combined surgical therapy for calculi of intrahepatic duct.</p><p><b>METHODS</b>One hundred and eight cases of hepatolithiasis treated in our hospital from January 1986 to September 2003 were summarized and analyzed retrospectively. The primary surgery included 57 cases of biliary tract exploration and cholangiolithotomy, 26 cases of cholangiojejunostomy, and 25 cases of partial hepatectomy. Of these cases, 156 operations were performed totally. There were 36 patients undergoing two or more operations for cholangiolithiasis problem. Fiberoptic choledochoscopy was employed to examine and remove the residual stones during and after operation in all cases.</p><p><b>RESULTS</b>The residual stone rates were 57.89% (33/57), 26.92% (7/26), and 24.0% (6/25) for biliary tract exploration and cholangiolithotomy, cholangiojejunostomy, and partial hepatectomy, respectively. Fiberoptic choledochoscopy was utilized pre- and post-operatively for each patient, which was performed 2 to 9 times on each case. Following the combined treatment, the rate of residual stone after operation decreased to 12.96% (14/108).</p><p><b>CONCLUSIONS</b>The combined surgical therapy is valuable for resolving the problem of hepatolithiasis in most of the patients. Regarding the clinical outcomes of different surgical procedures, partial hepatectomy is superior to cholangiojejunostomy or biliary tract exploration and cholangiolithotomy. Fiberoptic choledochoscopy is also important to reduce the occurrence of residual stones and the rate of reoperation.</p>

Risk factors for postoperative pulmonary complications after gastroduodenal operation / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the risk factors for postoperative pulmonary complications (PPC) after gastroduodenal operation.</p><p><b>METHODS</b>From December 1999 to December 2003, clinical data of 508 patients undergoing gastroduodenal operation were analyzed retrospectively. Risk factors for PPC were screened.</p><p><b>RESULTS</b>The complication rate of PPC was 25.8% (131/508). Multivariate logistic regression analysis revealed that age (OR=1.052), history of respiratory diseases (OR=2.915), serum albumin level (OR=0.995), length of intratracheal intubation (OR=1.005), length of nasogastric intubation (OR=1.059) and length of postoperative mechanical ventilation (OR=1.367) were risk factors for PPC.</p><p><b>CONCLUSION</b>Patients with old age, lower serum albumin level, intraoperative or postoperative nasogastric intubation, intratracheal intubation or long-term mechanical ventilation were more prone to develop PPC.</p>

Ventricular Endoscopy for Obstructive Hydrocephalus: Report of 33 Cases / 中国微创外科杂志

Objective To discuss the techniques of ventricular endoscopy for obstructive hydrocephalus.Methods A total of 33 patients with obstructive hydrocephalus were treated by ventriculostomy using ventricular endoscopy from June 2003 to December 2006.The patients had received CT,MRI,and 99mTc-DTPA cistern SPECT before the surgery.Results Endoscopic third ventriculostomy was performed successfully in 32 patients,the other patient died of hemorrhage under the third ventricle fistula.Postoperative fever occurred in 13 patients.One patient developed bilateral chronic subdural hematoma 6 months after the operation,and was cured by drill and drainage.Three patients developed subdural hydroma 6 months postoperation without showing symptoms.They were monitored without any treatment.The 32 patients were followed up for 2-42 months(mean,16.7 months),all of them achieved clinical relieve.In the patients,the intracranial pressure(ICP) was 26(16-39) cm H2O before the operation,and was significantly decreased to 22(12-28) cm H2O 7 days after the operation(t=2.903,0.005