High-level secretive expression of a novel achieved Talaromyces cellulolyticus endo-polygalacturonase in Pichia pastoris by improving gene dosage for hydrolysis of natural pectin.

Pectin is a type of complex hydrophilic polysaccharide widely distributed in plant resources. Thermal stable pectinase has its advantage in bioapplication in the fields of food processing, brewing, and papermaking, etc. In this study, we enzymatically characterized a putative endo-polygalacturonase TcPG from a Talaromyces cellulolyticus, realized its high-level expression in Pichia pastoris by in vitro constructing of a series of multi-copy expression cassettes and real time quantitative PCR screening. The secretive expression level of TcPG was nonlinear correlated to the gene dosage. Recombinants with five-copy TcPG gene in the host genome showed the highest expression. After cultivation in a bioreactor for about 96 h, the enzyme activity reached 7124.8 U/mL culture. TcPG has its optimal temperature of 70 °C. Under the optimized parameters, the pectin could be efficiently hydrolyzed into oligosaccharides.

Nano-microcapsule basic fibroblast growth factor combined with hypoxia-inducible factor-1 improves random skin flap survival in rats.

The present study aimed to investigate the effect of nano-microcapsule-basic fibroblast growth factor (bFGF) combined with hypoxia-inducible factor-1 (HIF-1) on the random skin flap survival of rats. Male Sprague-Dawley rats were used to establish the McFarlane flap model and subsequently, all model rats were randomly divided into four groups: Control, bFGF, HIF-1 and bFGF combined with HIF-1. The model rats were treated with 2.5 µg/day bFGF and 1.0 µg/day HIF-1 for 5 days by intraperitoneal injection. On day 5 following treatment, the boundaries between necrotic and surviving regions were significantly inhibited by bFGF combined with HIF-1. bFGF combined with HIF-1 inhibited oxidative stresses and inflammatory factors in random skin flap survival of rats. bFGF combined with HIF-1 also activated the protein expression levels of cyclooxygenase (COX)-2 and vascular endothelial growth factor (VEGF) in the random skin flap survival of rats. In conclusion, nano-microcapsule bFGF combined with HIF-1 prevented random skin flap survival in rats through antioxidative, anti-inflammatory and activation of the protein expression levels of COX-2 and VEGF.

Extramedullary versus intramedullary tibial alignment technique in total knee arthroplasty: A meta-analysis of randomized controlled trials.

The aim of this study was to establish whether the use of an extramedullary or intramedullary tibial cutting guide leads to superior mechanical leg axis and implant positioning. A meta-analysis of six randomized controlled trials including 350 knees was performed. For the mechanical axis, frontal tibial component angle and tibial slope, there were no significant differences in the mean values or the number of outliers (±3°) between the extramedullary and intramedullary groups. A reduced tourniquet time was associated with the intramedullary guide. No significant difference in the complication rate was noted between the two groups. Neither extramedullary nor intramedullary tibial alignment was more accurate in facilitating the tibial cut. Use of an intramedullary guide results in a shorter tourniquet time and exhibits a similar complication rate as the extramedullary guide.

Extramedullary versus intramedullary tibial alignment technique in total knee arthroplasty: A meta-analysis of randomized controlled trials

The aim of this study was to establish whether the use of an extramedullary or intramedullary tibial cutting guide leads to superior mechanical leg axis and implant positioning. A meta-analysis of six randomized controlled trials including 350 knees was performed. For the mechanical axis, frontal tibial component angle and tibial slope, there were no significant differences in the mean values or the number of outliers (±3°) between the extramedullary and intramedullary groups. A reduced tourniquet time was associated with the intramedullary guide. No significant difference in the complication rate was noted between the two groups. Neither extramedullary nor intramedullary tibial alignment was more accurate in facilitating the tibial cut. Use of an intramedullary guide results in a shorter tourniquet time and exhibits a similar complication rate as the extramedullary guide.

Hemodynamic Changes in Patients Undergoing Pericardiectomy.

OBJECTIVE: To observe the hemodynamic changes in patients undergoing pericardiectomy at different operational stages. METHODS: Totally 16 consecutive patients receiving radical pericardiectomy were enrolled in this observational study. Hemodynamic variables were monitored continuously by pulse-indicated continuous cardiac output(PiCCO)system. Totally,three sets of intraoperative hemodynamic parameters were obtained at three different stages of pericardiectomy. RESULTS: During the pericardiectomy,the cardiac index[CI,(1.9±0.6),(2.7±0.6),(3.0±0.5)L·min(-1)·m(-2);P<0.05]and stroke volume index[SI,(22.5±8.7),(29.9±8.5),(30.1±8.5)dyn·s·cm(-5)·m(2);P<0.05]showed significant improvement,whereas central venous pressure[CVP,(17.1±5.0),(13.3±3.9),(12.3±3.0)mmHg;P<0.05]decreased significantly. Global end-diastolic volume index[GEDVi,(533±156),(580±153),(559±144)ml·m(-2);P<0.05]increased and stroke volume variation[SVV,(15.6±6.1)%,(10.8±4.2)%,(9.4±5.4)%;P<0.05]decreased intra-operatively. The majority of the above-mentioned hemodynamic improvements occurred after the resection of pericardium over the left ventricular outflow tract(LVOT). CONCLUSIONS: PiCCO system can serve as a reliable,less invasive hemodynamic monitoring method during pericardiectomy. Resection of the pericardium over the LVOT is the most important step of the pericardiectomy.

Relationships between genetic polymorphisms in inflammation-related factor gene and the pathogenesis of nasopharyngeal cancer.

Our study aims to discuss the association between inflammation-related factors such as single nucleotide polymorphisms (SNPs) with susceptibility and recurrence in nasopharyngeal carcinoma. We used Taqman real-time polymerase chain reaction (PCR) to characterize the genetic variation of five SNPs in 194 nasopharyngeal carcinoma patients and 231 healthy subjects. All statistical analysis is performed with statistical product and service solutions v13.0; odds ratio (OR) value and 95 % confidence interval (CI) were calculated. There is no relationship between TGFß1 -869 T/C, IL-6 -634C/G, TGFß1 -509C/T, IL1 -511C/T and nasopharyngeal carcinoma susceptibility. Both single factor and multiple factors analysis showed that IL1a -889 T/T genotype is significantly associated with nasopharyngeal carcinoma in decreasing the risk of nasopharyngeal carcinoma. A highly significant association was found between IL1a -889 T/T genotype and protective genotype as defined by various pathological types. This is more obvious in the protective genotype of the non-keratin-type squamous carcinoma undifferentiated type. We also discovered that genotype G/G and C/G + G/G of IL6 -634 gene are associated with reduced recurrence of nasopharyngeal carcinoma. IL1a -889 gene polymorphism and susceptibility is related to nasopharyngeal carcinoma and can potentially decrease the risk of nasopharyngeal carcinoma in the Han Chinese population in north China. IL1-889 TT genotype is protective genotype for nasopharyngeal carcinoma. We have provided evidence that the GG genotype of the IL6 -634 gene is associated with recurrent risk of nasopharyngeal carcinoma. The G allele is the protective gene of nasopharyngeal carcinoma recurrence.

[Case-control study on therapeutic effects of different fixation methods for the treatment of olecranon fracture].

OBJECTIVE: To evaluate the clinical effects by comparing three different fixation methods: tension band, hollow lag screw and anatomical plate. METHODS: From January 2010 to January 2012, 82 patients with olecranon fractures who underwent surgical treatments were followed-up. All the patients were divided into three groups: tension band fixation group (group A), hollow lag screw fixation group (group B), anatomical plate fixation (group C). In group A, there were 35 patients, including 19 males and 16 females, ranging in age from 32 to 49 years old, with an average of (43.6 ± 8.7) years old, and the patients were treated with tension band fixation. According to Colton classification, there were 5 cases of type I, 3 cases of type II A,19 cases of type II B, and 8 cases of type II C in group A. Among 20 patients in group B, there were 13 males and 7 females, ranging in age from 27 to 50 years old, with an average of (41.5 ± 9.3) years old. The patients in group B were treated with hollow lag screw fixation. According to Colton classification, there were 4 cases of type I, 4 cases of type II A, and 12 cases of type II B in group B. In group C, there were 27 patients totally, including 15 males and 12 females, ranging in age from 30 to 55 years old, with an average of (38.2 ± 6.2) years old. The patients in group C were treated with anatomical plate fixation. According to Colton classification, there were 4 cases of type II B, 13 cases of type II C, and 10 cases of type II D in group C. The Fracture healing time, complications and functional recovery were retrospectively observed and recorded. RESULTS: All the patients were followed up, and the duration ranged from 8 to 24 months, with an average of 15 months. The average healing time of patients in group C was the longest among three groups. The flexion-extension and rotation activities of elbow joint in group B and C were better than that in group C. According to Broberg & Morrey score system, the therapeutic effects of patients in group A and B were better than that of group C. In group C, 2 patients had incision infections, 6 patients complained of foreign body sensation, 1 patient got a delayed fracture healing, and 1 patient had the heterotopic ossification. There were no occurrences of incision infections in group A and B; internal fixation loosening occurred in 3 patients in group A and 2 patients in group B; delayed fracture healing occurred in 2 patients in group A and 2 patients in group B; and skin bursa formation occurred in 6 patients in group A and 1 patient in group B. CONCLUSION: All the three ways are effective methods for the treatment of olecranon fractures. Fixation methods should be selected depending on the type of fracture.

[Prospective study on magnetic resonance-guided focused ultrasound surgery for symptomatic uterine fibroid: short-term follow up].

OBJECTIVE: To evaluate the safety and efficacy of magnetic resonance-guided focused ultrasound surgery (MRgFUS) in treatment of symptomatic uterine leiomyoma among Chinese reproductive age women. METHODS: From April 2010 to April 2012, 80 premenopausal women with symptomatic leiomyoma volunteered to participate in this prospective study in Department of Outpatient of Peking Union Medical College Hospital. Among 23 reproductive aged patients with size of uterus less than 16th gestational weeks, 2.5 to 10 cm of diameter of myoma, less than 10 myomas and expressing symptoms clearly were treated by MRgFUS. Treatment data, non-perfused volume ratio (NPVR) and adverse events were recorded. After treatment, patients were followed up at 1 week, 1, 3, 6, 12 and 24 months, respectively. Patients at initial screening and each time of the follow-up filled out uterine fibroid symptoms quality of life (UFS-QOL), which include symptoms severity score (SSS) and health-related quality of life (HRQL). The volumes of leiomyoma and uterine were evaluated on MRI before and after the treatment (at 6 and 12 months, respectively). Before operation, routine blood test were performed on all patients, anemia patients at 3 months and 1 year after treatment were checked with blood test. RESULTS: (1) Treatment data and adverse events: the mean therapeutic temperature was (69 ± 7)°C, the mean treatment time was (144 ± 62) min, the mean NPVR was (62 ± 23)%. Adverse events included mild erythema(1/23), abdominal cramp (8/23), vaginal discharge (5/23), and leg numbness (4/23). (2) The rate of secondary surgery: one patient was treated by myoectomy and hysterectomy within one year following up and 4 patients chose surgical treatment during the second-year follow-up. (3) Volume change:the volumes of leiomyoma before the treatment and 6, 12 months after the treatment are 75.6(P25 = 43.8, P75 = 128.9), 52.3(P25 = 23.8, P75 = 111.2), 45.9(P25 = 26.3, P75 = 71.7) cm(3), respectively; and the volumes of uterine before the treatment and 6, 12 months after the treatment are 270.0 (P25 = 208.4, P75 = 390.3), 216.4 (P25 = 151.1, P75 = 290.0), 200.0 (P25 = 149.1, P75 = 267.6) cm(3), respectively. Both leiomyoma and uterine volumes decreased significantly after treatments (P < 0.01). (4) UFS-QOL change:the symptoms severity score (SSS) before the treatment and 3, 12 months after the treatment are (34 ± 13), (22 ± 11), (19 ± 12), which decreased significantly (P < 0.01). The health-related quality of life (HRQL) before the treatment and 3, 12 months after the treatment are (74 ± 15), (82 ± 13), (89 ± 10), which increased dramatically (P < 0.01). (5) Hemoglobin (HGB) change: eleven patients suffered from anemia before treatments, the mean HGB before treatment was (87 ± 6) g/L and were (106 ± 14) g/L 3 months after treatment, (112 ± 10) g/L 12 months after treatment. The HGB was increased significantly after treatments (P < 0.01). CONCLUSIONS: MRgFUS is a safe and effective non-invasive management for symptomatic uterine leiomyoma in short-term follow up. But there is additional treatment ratio after MRgFUS.

[Influence of different tranexamic acid administration methods during and after cardiac surgery on coagulation function and postoperative blood loss].

OBJECTIVE: To evaluate the influence of different tranexamic acid administration methods during and after cardiac surgery with cardiopulmonary bypass(CPB) on coagulation function and postoperative bleeding. METHODS: Patients undergoing elective cardiac surgery with use of CPB (n=60) were randomized in a double-blind fashion to one of two treatment groups:group A(n=30) , administered with tranexamic acid 10 mg/kg (intravenous injection slowly before skin incision) , followed by infusion of normal saline until postoperative 12 hours;and group B(n=30) , administered with tranexamic acid 10 mg/kg(intravenous injection slowly before skin incision) , followed by infusion of tranexamic acid 1 mg/(kg·h) until postoperative 12 hours. Hemoglobin, platelet count, and coagulation function were assessed before anesthesia induction, after surgery, 8am next day and 24 hours after surgery. Bleeding, allogeneic blood transfusion, and fluid infusion during the postoperative 24 hours were recorded. RESULT: No differences were found between groups in terms of coagulant function, postoperative bleeding, allogeneic blood transfusion, and fluid infusion(P>0.05) . CONCLUSION: Compared with intraoperative administration alone, prolonged treatment with tranexamic acid after cardiac surgery shows no advantage because it can not further improve coagulant function, reduce bleeding, or reduce allogeneic blood transfusion.

[Analgesic effect of calpain inhibitor ALLN on the zymosan-induced paw inflammatory pain and its effect on the expression of cyclooxygenase-2 in the spinal dorsal horn].

OBJECTIVE: To examine the analgesic effect of calpain inhibitor ALLN on the zymosan-induced paw inflammatory pain and its effect on the expression of cyclooxygenase-2 (COX-2) in the spinal dorsal horn. METHODS: Forty-eight Sprague-Dawley rats were equally divided into three groups: control group, sham-operated group, and zymosan group. According to Meller's method, zymosan (1.25 mg) was injected intraplantarly to induce paw inflammation in zymosan group; an equal volume of PBS was administered in the sham-operated group. Mechanical withdrawal threshold (MWT) and maximum thickness of paw were tested or measured before and 0.5, 1, 2, 4, 8, and 24 hours after injection. All rats were killed at different occasions following surgery to examine calpain activity in the spinal dorsal horn with Western blot analysis. Another sixty-four Sprague-Dawley rats were divided into three groups: sham-operated group, zymosan-induced paw inflammation with intraperitoneal dimethyl sulphoxide (DMSO) treatment group, and zymosan-induced paw inflammation with intraperitoneal calpain inhibitor ALLN treatment group. MWT and maximum thickness of paw were tested or measured before and 0.5, 1, 2, 4, 8, and 24 hours after injection. All rats were killed at different occasions following surgery to examine the COX-2 expression in the spinal dorsal horn with Western blot analysis. RESULTS: MWT significantly decreased in the rats with zymosan-induced paw inflammation, while the maximum thickness of paw significantly increased, compared with control and sham-operated rats (P < 0.05). Calpain in the ipsilateral spinal dorsal horn was dramatically activated after zymosan injection (P < 0.01). Intraperitoneal ALLN injection significantly increased zymosan-induced MWT and decreased paw edema at the same time points after zymosan injection compared with DMSO treatment group (P < 0.05). Meanwhile, calpain inhibitor ALLN treatment significantly decreased the COX-2 expression in the spinal dorsal horn compared with DMSO treatment (P < 0.01). CONCLUSION: Administration of calpain inhibitor ALLN is effective to attenuate zymosan-induced paw inflammatory pain. Calpain activation may be one aspect of the signaling cascade that increases the COX-2 expression in the spinal cord and contributes to mechanical hyperalgesia after peripheral inflammatory injury.

[Use of blood salvage machine in oncologic surgeries: a clinical observation of 13 cases].

OBJECTIVE: To evaluate the effects on blood sparing and risk of hematogenous tumor dissemination of the use of blood salvage machine in oncologic surgeries. METHODS: The clinical data of 13 patients, 6 with malignant tumors and 7 with benign tumors, who received the use of blood salvage machine during oncologic surgeries based on informed consent, were analyzed. RESULTS: In total 42,575 ml of packed red blood cells were collected during surgery with a mean value of 3 275 ml (400 - 1500 ml) per patient. The average amount of allogenic transfusion per patient was 1 530 ml (0 - 8,000 ml). The number of blood salvage machine use in oncologic surgeries accounted for 8.6% (13/152) of the total number of blood salvage machine use in surgeries in that period. The perioperative mortality rate of the oncologic surgeries with the use of blood salvage machine was 7.7% (1/13). The post-operative metastasis rate of liver and lung was 15.4% (2/13). CONCLUSION: The use of blood salvage machine during oncologic surgeries improves the blood sparing effect. However, it cannot be used routinely, since it may result in hematogenous tumor cell dissemination.

[Evaluation of tracheal intubation in critical SARS patients].

OBJECTIVE: To analyze tracheal intubation and respiratory treatment in the critical severe acute respiratory syndrome (SARS) patients. METHODS: Review and analyze tracheal intubation and respiratory treatment in critical SARS patients in intensive care unit (ICU). RESULTS: Three of thirteen patients had been intubated or received tracheotomy before they entered into ICU, the other patients received treatment of nasal cannula or oxygen mask. With the development of the disease, two patients had been intubated because of respiratory failure or tracheotomy. Tracheal intubation was twice made in two patients in order to replace tracheal tubes. CONCLUSIONS: The patient should be intubated or received tracheotomy if non-invasive respiratory support has no effect. Standard protection could protect medical staff from infection under tracheal intubation.