The 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H), originated from Escherichia coli, converts p-coumaric acid to caffeic acid. In order to improve the efficiency of caffeic acid biosynthesis, we engineered E. coli for overexpression of 4HPA3H. The high-density fermentation of the engineered E. coli was conducted in a 5 L bioreactor. Subsequently, the conditions for whole-cell biocatalysis were optimized. The dry cell weight of the 4HPA3H-expressed strain reached 34.80 g/L. After incubated in the bioreactor for 6 h, 18.74 g/L (0.85 g/(L·OD₆₀₀)) of caffeic acid was obtained, with a conversion rate of 78.81% achieved. To the best of our knowledge, the titer of caffeic acid is the highest reported to date. The high-density fermentation of E. coli for overexpression of 4HPA3H and the efficient biosynthesis of caffeic acid may facilitate future large-scale production of caffeic acid.
Caffeic Acids ; Escherichia coli/metabolism* ; Fermentation ; Metabolic Engineering ; Mixed Function Oxygenases/metabolism* ; Phenylacetates

Traumatic rib fractures are the most common injury in thoracic trauma. Previously，the patients with traumatic rib fractures were mostly treated non-surgically，of which 50%，especially those combined with flail chest presented chronic pain or chest wall deformities and over 30% had long-term disabilities，being unable to retain a full-time job. In the past two decades，thanks to the development of internal fixation material technology，the surgical treatment of rib fractures has achieved good outcomes. However，there are still some problems in clinical treatment，including inconsistency in surgical treatment and quality control in medical services. The current consensuses on the management of regional traumatic rib fractures published at home and abroad mainly focus on the guidance of the overall treatment decisions and plans，and relevant clinical guidelines abroad lacks progress in surgical treatment of rib fractures in recent years. Therefore，the Chinese Society of Traumatology affiliated to Chinese Medical Association and Chinese College of Trauma Surgeons affiliated to Chinese Medical Doctor Association，in conjunction with national multidisciplinary experts，formulate the Chinese Consensus for Surgical Treatment of Traumatic Rib Fractures（2021）following the principle of evidence-based medicine，scientific nature and practicality. This expert consensus puts forward some clear，applicable，and graded recommendations from aspects of preoperative imaging evaluation，surgical indications，timing of surgery，surgical methods，rib fracture sites for surgical fixation，internal fixation methods and material selections，treatment of combined injuries in rib fractures，in order to provide references for surgical treatment of traumatic rib fractures.

Brain ; pathology ; Genome-Wide Association Study ; Humans ; Parkinson Disease ; genetics ; Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases ; genetics ; Proto-Oncogene Proteins c-crk ; genetics

OBJECTIVE： To investigate the effects of adiponectin （APN） on the expression of myocardial AMPK in myocardial insulin resistance （IR） model dogs during cardiopulmonary bypass （CPB）. METHODS： Totally 24 dogs were randomly divided into control group， model group， APN group （36 μg/kg）， AMPK inhibition group （APN 36 μg/kg+AMPK inhibitor compound C 0.5 mg/kg）， with 6 dogs in each group. All dogs underwent CPB； except for control group without medicine， CPB myocardial IR model were established in other groups， and perfused with St.Thomas cardiac cardioplegia lipid no medicine or containing relevant drugs after main artery block. Coronary sinus blood and carotid artery blood samples were collected before bypass and after 15， 90 min reperfusion following 60 min myocardial ischemia. Left ventricular apical tissue was taken， and the uptake rate of myocardial glucose and insulin resistance index （IRI） were determined and calculated； the changes of myocardial injury indexes （cTnT concentration） and cardiac function indexes （LVSP， +dp/dtmax） were monitored. The level of p-AMPK was detected. RESULTS： There was no statistical significance in above indexes of dogs before bypass （P＞0.05）. Compared with control group， the rate of myocardial glucose uptake， the levels of LVSP， +dp/dtmax and p-AMPK in model group were decreased significantly after 15， 90 min reperfusion （P＜0.05）， and the concentrations of IRI and cTnT were increased significantly （P＜0.05）. Compared with model group， the rate of myocardial glucose uptake， LVSP， +dp/dtmax and p-AMPK were increased significantly in APN group and AMPK inhibitor group （P＜0.05）， while the concentrations of IRI and cTnT were decreased significantly （P＜0.05）； moreover， the effect of APN group was better than that of AMPK inhibitor group （P＜0.05）. CONCLUSIONS： APN can promote myocardial glucose uptake and metabolism， and contribute the recovery of cardiac function， the mechanism of which may be associated with increasing the activity of AMPK.

Objective To investigate the safety and longterm outcomes of prophylactic autologous pericardium tricuspid valve annuloplasty(TVA)in patients with rheumatic heart disease(RHD). Methods A total of 832 patients with RHD were enrolled in this study ,including 146 patients with mild FTR but without TVA(observation group);434 patients with mild FTR underwent TVA(control group A)and 434 patients with moderate or severe FTR underwent TVA(control group B). Propensity score and survival analysis were used to evaluate perioperative safety ,FTR progression ,CHF and MACCE incidence after prophylactic TVA. Results A total of 192 patients were successfully matched. There was no significant difference in the perioperative complications between the observation group and the control groups(P>0.05). The progression rate of FTR in the observation group was significantly lower than that in the control groups (P = 0.005 & 0.032 ). There was no significant difference in the incidence of CHF and MACCE events between the observation group and the control groups (P > 0.05). Conclusions The treatment strategy of prophylactic autologous pericardium TVA at the time of left heart valve surgery for patients with RHD doesn′t increase operation costs ,perioperative complications and mortality ,but effectively prevent postoperative FTR recurrence or progression.

OBJECTIVE:To explore the effects of acadesine on myocardial energy metabolism of model dogs with myocardial ischemia-reperfusion injury (MIRI) after cardiopulmonary bypass (CPB). METHODS:Dogs were randomly divided into control group,model group,acadesine low-dose,high-dose groups(0.8,3.2 mg/kg),6 in each group. All dogs received CPB. Except for control group,dogs in other groups were reduced for MIRI model,and perfused St.Thomas cardiac cardioplegia lipid containing rel-evant drugs 60 min after main artery block. The uptake rates of myocardial glucose and free fatty acid(FFA),creatine kinase isoen-zyme(CK-MB)concent in venous sinus plasma and adenosine triphosphate(ATP)content in mitochondria were detected and calcu-lated before bypass and after 15,60,90 min of reperfusion. Left ventricular systolic pressure(LVSP)and left ventricular end dia-stolic pressure(LVEDP)were analyzed,and mRNA expression of adenylate-activated protein kinase(AMPK)and protein expres-sion of phosphorylated AMPK(p-AMPK)in myocardial tissue were detected. RESULTS:Before bypass,all indexes in each group had no statistic significances(P>0.05). After bypass,compared with control group,uptake rates of myocardial glucose and FAA, ATP content,mRNA expression of AMPK and protein expression of p-AMPK and LVSP in 3 time points in model group and each administration group were obviously decreased(P<0.05);LVEDP and CK-MB concent in plasma were obviously increased(P<0.05). Compared with model group,uptake rates of myocardial glucose and FAA,ATP content,mRNA expression of AMPK and protein expression of p-AMPK and LVSP in 3 time points in each administration group were obviously increased (P<0.05);LVEDP and CK-MB concent in plasma were obviously decreased (P<0.05);and high-dose group showed more obvious change than that of low-dose group (P<0.05). CONCLUSIONS:Acadesine can promote the AMPK phosphorylation,contribute to the myocardial glucose and FFA uptake to promote the increase of ATP in myocardial mitochondria and relieve MIRI after CPB.

Objective To compare the ear baric function between 4000m altitude chamber test with inhaling air and 6900m altitude chamber test with inhaling pure oxygen.Methods Eleven healthy male volunteers attended two tests as two groups by self-comparison. As the air group the volunteers inhaled air at 4000m, while as the pure oxygen group they inhaled pure oxygen at 6900m altitude, and the time interval between the two tests was more than two weeks. During the test, the volunteers breathed air or pure oxygen at random for 1h, and then were exposed at a speed of 20m/s to the target altitude for 5min. Hereafter they were sent back to the ground at the same speed. The changes of subjective symptoms, degree of tympanic congestion, acoustic immitance index and pure-tone auditory threshold were recorded before and after the test. The acoustic impedance index and pure-tone threshold were statistically analyzed.ResultsFour volunteers (4 ears) in air group and 7 volunteers (7 ears) in pure oxygen group reported ear pain in altitude chamber exposures, respectively. The pain-triggering altitude was higher in the pure oxygen group. Immediately after tests, there were 3 (3 ears) and 5 volunteers (5 ears) with Ⅲ degree congestion of the tympanic membrane in the two groups respectively. Four volunteers (6 ears) developed gradually aggravated hemorrhages after altitude exposure. And the tympanic membrane congestion difference between groups was statistically significant at 3 and 24h after tests (P<0.01). The type A tympanogram appeared in 11 (15 ears) and 11 (14 ears) volunteers respectively immediately after tests. The increase of static compliance value was significantly greater in pure oxygen group than in air group immediately after tests (P<0.05), the decrease of middle ear pressure was more significant in pure oxygen group than in air group at 3 and 24h after tests (P<0.05). Both the two altitude exposure tests resulted in eustachian tube dysfunction. At 3 and 24h after the tests, the increase of individual frequency pure-tone threshold was significantly higher in pure oxygen group than in air group (P<0.05).Conclusion Breathing pure oxygen and lifting height could increase the screening degree of ear baric function test in hypobaric chamber, and have greater influence on degree of tympanic congestion, acoustic immittance and pure-tone auditory threshold in 24 hours.

Objective: To summarize the methods of preventing and managing the complications in thoracoscopic lobectomy. Methods:The participants of this study included 317 patients undergoing lobectomy with video-assisted thoracoscopic surgery in the Department of Thoracic Surgery between January 2007 and December 2012. Intra-operative complications were observed, and countermeasures were summarized. Results: Complications occurred 28 times (8.8%), including bleeding in 16 cases because of accidental vascular injury (5.0%), accidental injury/break of bronchus in two cases (0.6%), vascular stump errhysis from cutting stapler in four cases (1.3%), lung stump air leakage in three cases (0.9%), lung injury in two cases (0.6%), and diaphragmatic injury in one case (0.3%). Conversion to thoracotomy was conducted in 17 cases, with a conversion rate of 5.4%. Thoracoscopic repair operation was performed in 14 cases that exhibited bleeding, with a success rate of 70% (14/20). No mortality was reported during the operation. Conclusion:Thoracoscopic lobectomy is a highly difficult method in thoracic surgeries. The procedure requires substantial attention on the timely prevention and correct management of intra-operative complications, particularly the injury and bleeding of major vessels, to reduce the rate of conversion to thoracotomy and the incidence of post-operative complications, as well as to promote the surgery in clinics.

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.

This study aims to investigate the effects of fibroblast growth factor 21 (FGF-21) on learning and memory abilities and antioxidant capacity of D-galactose-induced aging mice. Kunming mice (37.1 +/- 0.62) g were randomly divided into normal control group, model group and FGF-21 high, medium and low dose groups (n = 8). Each group was injected in cervical part subcutaneously with D-galactose 180 mg x kg(-1) x d(-1) once a day for 8 weeks. At the same time, FGF-21-treated mice were administered with FGF-21 by giving subcutaneous injection in cervical part at the daily doses of 5, 2 and 1 mg x kg(-1) x d(-1). The normal control group was given with normal saline by subcutaneous injection in cervical part. At seventh week of the experiment, the learning and memory abilities of mice were determined by water maze and jumping stand tests. At the end of the experiment, the mice were sacrificed and the cells damage of hippocampus was observed by HE staining in each group. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and total antioxidant capacity (T-AOC) in the brain of mice were determined. The results showed that different doses of FGF-21 could reduce the time reaching the end (P < 0.01 or P < 0.05) and the number of touching blind side (P < 0.01 or P < 0.05) in the water maze comparing with the model group. It could also prolong the latency time (P < 0.05) and decrease the number of errors (P < 0.01 or P < 0.05) in the step down test. The result of HE staining showed that FGF-21 could significantly reduce brain cell damage in the hippocampus. The ROS and MDA levels of three different doses FGF-21 treatment group reduced significantly than that of the model group [(5.58 +/- 1.07), (7.78 +/- 1.92), (9.03 +/- 1.77) vs (12.75 +/- 2.02) pmol (DCF) x min(-1) x mg(-1), P < 0.01 or P < 0.05], [(2.92 +/- 0.71), (4.21 +/- 0.81), (4.41 +/- 0.97) vs (5.62 +/- 0.63) nmol x mg(-1) (protein), P < 0.01]. Comparing with the model group, the activities of SOD, GPx, CAT and T-AOC of the three different doses FGF-21 treatment groups were also improved in a dose-dependent manner. This study demonstrates that FGF-21 can ameliorate learning and memory abilities of D-galactose induced aging mice, improve the antioxidant abilities in brain tissue and delay brain aging. This finding provides a theoretical support for clinical application of FGF-21 as a novel therapeutics for preventing aging.