Geminiviruses cause substantial crop yield losses worldwide. The replication initiator protein (Rep) encoded by geminiviruses is indispensable for geminiviral replication. The Rep protein encoded by beet severe curly top virus (BSCTV, genus Curtovirus, family Geminiviridae) induces VARIANT IN METHYLATION 5 (VIM5) expression in Arabidopsis leaves upon BSCTV infection. VIM5 functions as a ubiquitination-related E3 ligase to promote the proteasomal degradation of methyltransferases, resulting in reduction of methylation levels in the BSCTV C2-3 promoter. However, the specific domains of Rep responsible for VIM5 induction remain poorly characterized. Although Rep proteins from several geminiviruses act as viral suppressors of RNA silencing (VSRs), whether BSCTV Rep also possesses VSR activity remains to be illustrated. In this study, we employed a transient expression system in the 16c-GFP transgenic and the wild-type Nicotiana benthamiana plants to analyze the VSR and the VIM5-inducing activities of different truncated Rep proteins haboring distinct domains. We found that the N-terminal domain (amino acids 1-180) of Rep suppressed GFP silencing in 16c-GFP transgenic N. benthamiana leaves. The minimal N-terminal fragment (amino acids 1-104) induced VIM5 expression upon co-infiltration, while C-terminal truncations lacked VIM5-inducing activity. Our results indicate that the N-terminal domain of Rep encoded by BSCTV mediates the suppression of RNA silencing and induces VIM5 expression. Thus, our findings contribute to a better understanding of interactions between geminiviral Rep and plant hosts.
Geminiviridae/genetics* ; Nicotiana/metabolism* ; Arabidopsis/metabolism* ; RNA Interference ; Viral Proteins/metabolism* ; Arabidopsis Proteins/metabolism* ; Plants, Genetically Modified/metabolism* ; Protein Domains ; Plant Diseases/virology* ; Methyltransferases/metabolism* ; Ubiquitin-Protein Ligases/metabolism* ; DNA Helicases/genetics*

Objective To determine the effect of isopsoralen(ISO)on the healing of tibia fracture in mice and explore its underlying mechanism.Methods Fifty male C57BL/6 mice(2 month old,20±2 g)were randomly divided into model group and ISO treatment group,with 25 animals in each group.From the 3rd day after modeling,the mice from the ISO group were given an intragastric gavage of 40 mg/kg ISO,once per day for 28 consecutive days,while those of the model group was given same volume of normal saline in same way.On the 7th,14th,21st,and 28th day after gavage,the tibia on the surgical side was taken,and the fracture area was quantified by bone volume/total volume(BV/TV)after micro-CT scanning.The healing and shaping of the fracture end were observed through HE staining.ELISA was used to detect the serum contents of bone alkaline phosphatase(BALP)and procollagen type I N-terminal peptide(PINP)on the 14th day of gavage.Western blotting was employed to determine the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX,and VEGF in the tibial callus tissue in 7 and 14 d after gavage.Vascular perfusion was applied to observe the callus microvessels in 28 d to quantitatively analyze the vascular volume fraction and vessel diameter.Immunohistochemical staining was conducted to observe the expression of VEGF in the callus in 14 d after gavage.Results HE staining displayed that the ISO group had faster healing process than the model group.Micro-CT quantification results showed that the ISO group had higher BV/TV ratio in 7 d after gavage though no statistical difference,significantly higher ratio in 14 d(P＜0.05),but obviously lower ratio in 21 and 28 d after gavage(both P＜0.05)when compared with the model group.The serum contents of BALP and PINP were also remarkably higher in the ISO group than the model group(P＜0.05).Western blotting results indicated that the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX and VEGF in the ISO group were higher than those in the model group(P＜0.05).The results of angiography revealed that the vascular volume fraction and vessel diameter were notably increased in the ISO group than the model group(both P＜0.05).Immunohistochemical assay showed that the expression of VEGF was higher in the ISO group than the model group(P＜0.05).Conclusion ISO can improve the activity of osteoblasts,increase the expression of osteogenesis-related proteins,and accelerate the angiogenesis to promote fracture healing.

BACKGROUND: Cholecystectomy is a standard surgery for patients suffering from gallbladder diseases, while the causal effects of cholecystectomy on colorectal cancer (CRC) and other complications are still unknown. METHODS: We obtained genetic variants associated with cholecystectomy at a genome-wide significant level ( P value <5 × 10 -8 ) as instrumental variables (IVs) and performed Mendelian randomization (MR) to identify the complications of cholecystectomy. Furthermore, the cholelithiasis was also treated as the exposure to compare its causal effects to those of cholecystectomy, and multivariable MR analysis was carried out to judge whether the effect of cholecystectomy was independent of cholelithiasis. The study was reported based on Strengthening the Reporting of Observational Studies in Epidemiology Using Mendelian Randomization guidelines. RESULTS: The selected IVs explained 1.76% variance of cholecystectomy. Our MR analysis suggested that cholecystectomy cannot elevate the risk of CRC (odds ratio [OR] =1.543, 95% confidence interval [CI]: 0.607-3.924). Also, it was not significant in either colon or rectum cancer. Intriguingly, cholecystectomy might decrease the risk of Crohn's disease (OR = 0.078, 95% CI: 0.016-0.368) and coronary heart disease (OR = 0.352, 95% CI: 0.164-0.756). However, it might increase the risk of irritable bowel syndrome (IBS) (OR = 7.573, 95% CI: 1.096-52.318). Cholelithiasis could increase the risk of CRC in the largest population (OR = 1.041, 95% CI: 1.010-1.073). The multivariable MR analysis suggested that genetic liability to cholelithiasis could increase the risk of CRC in the largest population (OR = 1.061, 95% CI: 1.002-1.125) after adjustment of cholecystectomy. CONCLUSIONS The study indicated that cholecystectomy might not increase the risk of CRC, but such a conclusion needs further proving by clinical equivalence. Additionally, it might increase the risk of IBS, which should be paid attention to in clinical practice.
Humans ; Mendelian Randomization Analysis ; Irritable Bowel Syndrome ; Colorectal Neoplasms/genetics* ; Cholelithiasis/complications* ; Cholecystectomy/adverse effects* ; Genome-Wide Association Study ; Polymorphism, Single Nucleotide

Objective:To study the effects of a simulated plateau environment on fracture healing in rats.Methods:A rat model of mid-femoral fracture was established by hacksaw truncation and intramedullary fixation with Kirschner wires in 60 male Wistar rats which were divide into 2 groups ( n=30) by the random number table method. The rats in the control group were raised in the animal experiment center of The 940 Hospital of Joint Logistic Support Force of Chinese PLA at an altitude of 1,400 m, while the rats in the plateau group were placed in an animal experimental cabin in a simulated plateau environment at a simulated altitude of 5,000 m. The body weight was weighed once a week and X-ray films were taken every 2 weeks. Blood samples were collected after 4 weeks for detection of biochemical indicators of bone metabolism. After 8 weeks, the femurs of the surgical side were taken for bone biomechanical detection and the bone mineral density of the healthy side was detected. After 4 and 8 weeks, the femurs of the surgical side were taken for in vitro Micro-CT scanning and angiography detection. After 1, 2, 4 and 8 weeks, the femurs of the surgical side were taken for bone histopathologic detection. Results:During the entire experiment, no rats in the control group died while the mortality rate of the rats in the plateau group was as high as 26.7% (8/30). In the plateau group, some organs were pathologically damaged in the rats, fracture union was delayed, and the callus differentiated and matured slowly with the chondrocytes still dominant at the 8th week. The bone mineral density and the maximum load of the femur in the plateau group were significantly lower than those in the control group ( P< 0.05). Angiography showed that the rats in the plateau group had microvascular proliferation which did not penetrate the fracture end at the 8th week. The bone formation indexes like osteocalcin, procollagen type Ⅰ N-terminal propeptide (PⅠNP), and osteoprotegerin of the rats in the plateau group were significantly lower than those in the control group at the 4th week ( P<0.05). The bone resorption indexes like tartrate resistant acid phosphatase 5b (TRACP-5b) and receptor activator for nuclear factor-κB ligand (RANKL) in the plateau group were significantly higher than those in the control group ( P<0.05). Conclusion:A simulated plateau environment at an altitude of 5,000 m may lead to delayed fracture healing in rats.

Objective:To investigate the diagnosis and treatment of emergency inguinal hernia.Methods:The retrospective cross-sectional study was conducted. The clinical data of 236 patients with emergency inguinal hernia who were admitted to the First Affiliated Hospital of Soochow University from January 2015 to May 2020 were collected. There were 194 males and 42 females, aged (69±30)years. Hospitalized patients received routine blood biochemistry test and imaging examinations for evaluation of characteristics of hernia contents and intestinal obstruction. Manual reduction and surgical treatment were selected according to the conditions of patients. Observation indicators: (1) treatment; (2) follow-up. Follow-up using outpatient examination and telephone interview was performed to detect hernia recurrence and late-onset mesh infection up to August 2020. Measurement data were described as M (range) or M ( P25, P75), and comparison between groups was analyzed using the Wilcoxon rank sum test. Count data were represented as absolute numbers, and comparison between groups was done using the chi-square test. Results:(1) Treatment: of the 236 patients, 106 cases had successful manual reduction, 124 cases underwent emergency operation, 6 cases refused surgery. ① For 106 cases with successful manual reduction (including 4 cases guided by B-ultrasonography), the manual reduction time was 5 minutes (2 minutes,7 minutes). Ninety-three of 106 patients underwent selective operation after manual reduction, including 89 cases with indirect hernia, 2 cases with direct hernia and 2 cases with compound hernia. The time to selective operation was 3 days(2 days,5 days) after manual reduction. Patients underwent mesh repair, of which the operation time, volume of intraoperative blood loss, time to postoperative first flatus, duration of postoperative hospital stay were 44 minutes (29 minutes, 66 minutes),10 mL(5 mL,20 mL), 1 day(1 day,2 days), 1 day(1 day,2 days), respectively. Eleven patients didn't undergo selective operation. Two patients with abdominal pain and fever after manual reduction were diagnosed with perforation of intestine by emergency surgical exploration, and then underwent partial intestinal resection combined with high ligation of hernial sac. ② There were 93 of 124 patients undergoing emergency operation with indirect hernia, 18 cases with femoral hernia, 6 cases with obturator hernia, 6 cases with compound hernia and 1 case with direct hernia. There were 54 of 124 patients undergoing open operation, including 21 cases with Bassini surgery, 18 cases with Lichtenstein surgery, 9 cases with Mc Vay surgery, 6 cases with high ligation of hernia sac. There were 70 patients undergoing laparoscopic operation, including 57 cases with laparoscopic transperitoneal preperitoneal hernia repair (TAPP), 10 cases with laparoscopic explora-tion + tissue repair and 3 cases with laparoscopic exploration + closure of inner inguinal ring. The operation time, volume of intraoperative blood loss, time to postoperative first flatus, cases with short-term postoperative complications were 60 minutes (50 minutes,76 minutes), 20 mL(14 mL,30 mL), 2 days(1 day,2 days), 15 cases for patients undergoing open surgery, respectively. The above indicators were 56 minutes (47 minutes,77 minutes), 20 mL(10 mL,25 mL), 2 days(1 day,2 days), 21 cases for patients under-going laparoscopic surgery. There was no significant difference in the above indicators between the two groups ( Z=?0.88, ?1.37, ?1.56, χ2=0.07, P>0.05). Cases with intraoperative placement of mesh and duration of hospital stay were 18 cases and 5 days(3 days,8 days) for patients undergoing open surgery, versus 57 cases and 3 days(2 days,5 days) for patients undergoing laparoscopic surgery, showing significant differences between the two groups ( χ2=29.50, Z=?4.32, P<0.05). (2) Follow-up: of 236 patients, 192 were followed up for 2?60 months, with a median follow-up time of 19 months. Seven patients had recurrence of hernia after emergency operation, including 3 with high ligation of the hernia sac, 2 with Bassini surgery, 1 with Lichtenstein surgery, and 1 with laparoscopic exploration + closure of inner inguinal ring. One patient with late-onset mesh infection after Lichtenstein surgery was improved after mesh removal. No long-term complications such as hernia recurrence or late-onset mesh infection occurred to the 184 patients. Conclusions:Emergency inguinal hernia had different state of illness, manual reduction is suitable for partial patients with incarceration. Surgery is the first choice, and the surgical procedure needs to be individually selected.

The pathophysiological process of immune inflammatory response after severe trauma is extremely complex, especially manifested in the dynamic changes. In the physiological response state, the inflammatory and anti-inflammatory conditions are in a dynamic balance. The immune inflammatory response is relatively stable, avoiding excessive inflammatory reactions or immunosuppression and reducing further damage to the body. In the pathological response state, the dynamic balance between inflammatory and anti-inflammatory is broken, and it can also lead to persistent inflammatory-immunosuppression-catabolism syndrome (PICS). As a result, it increases serious complications such as uncontrolled inflammatory reactions, sepsis, multiple organ dysfunction syndrome (MODS), and multiple organ failure (MOF). Current researches on post-traumatic immune inflammatory response have also expanded to the genetic level, indicating that the over-expression of genes and the generation and increase of immune response media are likely to be the key reasons for the disorder of immune inflammatory response. The author reviews the research progress of immune inflammatory response mechanism and related clinical intervention after severe trauma, in order to summarize the previous research results and explore the future research direction.

Objective To observe the effect of stress on the rapid component of delayed rectifier potassium current (IKr) in rat cardiomyocytes. Methods Forty male SD rats were randomly divided into four groups (10 each): control group (Ctrl), exhaustive group (ES), noise group (WN) and composite group (ES+WN). Stress animal models were prepared as follows: Rats in ES group were undergoing exhaustive swimming as the stress factor, in WN group undergoing white noise and in ES+WN group undergoing exhaustive swimming + white noise as the stress factor. Langendorff device was used to reversely perfuse collagenase for isolating the rat's ventricular myocytes. The effect of stress on IKr current and gating mechanism of single ventricular myocyte was recorded by whole-cell patch clamp technique. Results Compared with the Ctrl group, the tail current density of IKr (IKr,tail) of ventricular myocytes increased significantly in ES group and WN group (P<0.01). The IKr,tail current density of the ventricular myocytes in ES+WN group was significantly higher than that in ES group and WN group (P<0.01), and the effect was voltage dependent. Gating mechanism revealed that the half inactivation voltage of IKr,tail (V1/2,inact) can be shifted to the right in ES group, WN group and ES+WN group when compared with the Ctrl group, and the recovery time constant shortened after inactivation (P<0.01). However, the steady-state activation, fast inactivation constant and voltage dependence of IKr,tail were not statistically significant in ES group, WN group and ES+WN group when compared with the Ctrl group. Conclusion Stress increases the IKr current in rat cardiomyocytes, suggesting it be one of the electrophysiological bases of stress-induced arrhythmia.

Objective To evaluate the feasibility and outcome of laparoscopic cholecystectomy as a overnight procedure in China.Methods The data of 59 consecutive patients who had undergone outpatient laparoscopic cholecystectomy between Januaiy 2013 and January 2015.All the patients were operated in the morning hours and discharged during 24 hours after operation.They were contacted by telephone 3 days subsequent to surgery and were seen in the outpatient unit 7 days after.Results Fifty-nine laparoscopic cholecystectomies were performed.No Conversion to open surgery case.The average operation time was 25 minutes,and restore semi-liquid diet 6 hours after the operation All the 59 patients were discharged during 24 hours after operations.None of the patients had an emergency readmission.None of the patients had complications 7 days after discharged.The average payment was 8 240 yuan.Conclusion These results suggest that laparoscopic cholecystectomy can be routinely performed as a overnight procedure.

Objective To study the effect of allicin ( All) on potassium current in single atrial myocytes in rats. Methods Isolated rat atrial myocytes were isolated by perfusion and administered by extracellular perfusion method.Whole cell patch clamp technique was used to record transient outward potassium current ( Ito ) in atrial myocyte. Results In presence of 30 μmol·L-1 of All,the peak current of Ito was significantly reduced from (20.5±2.2) pA/pF to (11.3±2.1) pA/pF at+50 mV of test potential (P<0.01,n=12).This effect of All showed voltage- and concentration-dependence with IC50 of (19.0±2.5)μmol·L-1 .The steady-state activation curve of Ito was shifted to more positive potential and recovery time from inactivation was prolonged.In addition, they fail to find any effect of All on the steady-state inactivation and closed-state inactivation of Ito. Conclusion All can inhibit Ito by slowing down the process of activation and recovery from inactivation of channel.