Objective To explore the rehabilitation for amputation and paraplegia. Methods A case with both lower limbs and left arm amputation complicated with L1 complete spinal cord injury was reported. Results Her participation improved after rehabilitation. Conclusion It is important to improve the participation under the limit of activity.

Objective To construct lentivirus vectors carrying 16 short hairpin RNA (shRNA) expression cassettes targeting histone acetyltransferases and provide a powerful research approach to explore the mechanism of epigenetic genes in regulating hepatitis B virus (HBV).Methods Following the rule of short shRNA primer design,eight-pair primers (A ~ H )for each gene,which had stable interfering efficiency,were designed.The annealed primers were connected to the empty lentiviral vectors of shRNA for transformation.In order to confirm the positive clones,clones were analyzed by real-time polymerase chain reaction (RT-PCR ).Then, qualified plasmids were verified by enzyme digestion technology.Four shRNA lentivirus plasmids against the same gene were mixed to build lentivirus respectively.After the virus transfected into 293T cells for 48 and 72 hours,supernatants were collected to infect HepG2.2.15 cells.The percentage of fluorescent cells were observed and assessed by microscope 72 hours after infection.Results One hundred and twenty-eight lentiviral vectors of RNA interference (RNAi)library were constructed against 16 histone acetyltransferases and more than 80% of HepG2.2.15 cells were infected with lentivirus 72 hours after infection.Conclusions Sixteen shRNA lentivirus vectors against histone acetyltransferase are successfully constructed.Thus,a solid foundation for the study of the effect of human histone deacetylase on HBV replication is established.

The in vitro release is an important index to evaluate the quality of liposome formulation.Currently, there is no evaluation method for the in vitro release of liposome in pharmacopoeia of various countries, which leads to the lack of unified standard and safety guarantee for the quality evaluation of liposome formulation.Taking the self-made paclitaxel derivative liposomes as an example, the paddle membrane binding method established by optimizing external release conditions was used to simulate the complete release of paclitaxel derivative drugs in 12 hours under physiological conditions.The results showed that using 0.5% SDS-HEPES as the release medium and a dialysis bag with a molecular weight cutoff of 1 000 kD to release the liposome solution met the requirements and had discrimination ability, providing a reference for the development of drug-loaded liposomes release methods in vitro.

Objective:To evaluate the clinical value of the transcutaneous neuromodulation (TN) in improving gastrointestinal function after gastrointestinal tumor operation.Methods:From April 2019 to June 2020, at The Affiliated People′s Hospital of Ningbo University, 100 patients who underwent gastrointestinal tumor surgery were included. The 100 patients were randomly divided into treatment group(receiving TN treatment, 50 cases)and control group (receiving sham TN treatment, 50 cases). The clinical data of the two groups was compared to evaluate the recovery of gastrointestinal function, which included the time of first defecation, time of first flatus, time of first ambulation, time of resuming diet, the incidence of nausea and vomiting within 3 d after operation and pain score (0 to 10). Heart rate variability (HRV) was compared between two groups to analyze the possible mechanism of TN improving gastrointestinal function after gastrointestinal tumor surgery. Independent sample t test and Chi-square test were used for statistical analysis. Results:Among 100 patients, there were 63 male and 37 female patients, the age was (67.0±11.3) years old, ranged from 28 to 92 years old. Compared with the control group, the time of first defecation, first flatus, first ambulation and resuming diet of treatment group reduced by 31.0%, 39.8%, 38.0% and 32.4% ((72.1±3.0) h vs.(104.5±2.9) h, (49.4±5.7) h vs.(82.1±3.1) h, (3.1±0.7) d vs.(5.0±0.9) d, (4.8±0.9) d vs. (7.1±0.8) d)), respectively; the pain scores on the day 2 and day 3 after operation and incidence of nausea and vomiting within 3 d after operation decreased by 50.0%, 65.5%, 26.0%(1.5±0.6 vs. 3.0±0.7, 1.0±0.6 vs. 2.9±0.6, 16.0%, 8/50 vs. 42.0%, 21/50), respectively, and the differences were statistically significant ( t=54.28, 35.72, 11.67, 13.66, 12.00 and 14.90, χ2=8.21, all P<0.01). The results of HRV analysis showed that the high frequency on day 3 was higher than that on day 1 of treatment group, and the ratio of low frequency to high frequency after operation was lower than that before operation of treatment group (0.5±0.1 vs. 0.4±0.1, 1.2±0.7 vs. 1.9±1.0), and the differences were statistically significant( t=-4.81 and 4.26, both P<0.01), which indicated TN could enhance vagal activity. Conclusions:TN promote the recovery of gastrointestinal function after gastrointestinal tumor operation, and can be used as an adjuvant therapy to accelerate the recovery of gastrointestinal function after gastrointestinal tumor operation.

Current in vitro assays for the activity of HIV-RT (reverse transcriptase) require radio-labeled or chemically modified nucleotides to detect reaction products. However, these assays are inherently end-point measurements and labor intensive. Here we describe a novel non-radioactive assay based on the principle of pyrosequencing coupled-enzyme system to monitor the activity of HIV-RT by indirectly measuring the release of pyrophosphate (PP(i)), which is generated during nascent strand synthesis. The results show that our assay could monitor HIV-RT activity with high sensitivity and is suitable for rapid high-throughput drug screening targeting anti-HIV therapies due to its high speed and convenience. Moreover, this assay can be used to measure primase activity in an easy and sensitive manner, which suggests that this novel approach could be wildly used to analyze the activity of PP(i)-generated and ATP-free enzyme reactions.
Anti-HIV Agents ; pharmacology ; Colorimetry ; Diphosphates ; analysis ; metabolism ; Drug Evaluation, Preclinical ; HIV ; drug effects ; enzymology ; HIV Reverse Transcriptase ; analysis ; antagonists & inhibitors ; metabolism ; Humans ; In Vitro Techniques ; Nevirapine ; pharmacology ; Reverse Transcriptase Inhibitors ; pharmacology ; Sequence Analysis, DNA ; Thymine Nucleotides ; metabolism

Surgical operation is the main treatment method for pancreatic cancer, and in clinical practice, radical surgery for pancreatic cancer is often combined with superior mesenteric-portal vein confluence pancreaticoduodenectomy to achieve R0 resection. However, severe left-sided portal hypertension (LSPH) may occur after splenic vein dissection, resulting in a series of pathological changes such as congestive splenomegaly, thrombocytopenia, backflow obstruction of splenic vein, and gastrointestinal varices, and in some cases, it can lead to fatal gastrointestinal hemorrhage and hemorrhagic shock. Therefore, in order to better manage LSPH in clinical practice, this article systematically analyzes and reviews the pathogenesis, treatment regimens, and control strategies of LSPH after combined superior mesenteric-portal vein confluence pancreaticoduodenectomy and put forward corresponding suggestions based on current studies.

【Objective】 To prepare liposomes encapsulate hemoglobin and paclitaxel(LEHP)to improve tumor hypoxia resistance. 【Methods】 LEHP were prepared by thin-film method, and the particle size, Zeta potential and polydispersity were investigated by nanoparticle size analyzer, and encapsulation efficiency was investigated by high performance liquid chromatography, and the interaction between the liposomes and tumor cells was evaluated by in vitro cell experiments. 【Results】 The optimal preparation conditions of LEHP was as follows: total phospholipid 36 mM, DPPC∶Dope∶cholesterol molar ratio 7∶2∶1, paclitaxel 3 mg, hydrated with 3 mg·mL^-1 Hb-PBS for 30 min at room temperature; The average particle size was (189.17±8.22) nm, polydispersity was 0.14±0.023, paclitaxel encapsulation efficiency was (58.27±2.55)%, hemoglobin content was (0.63±0.05) mg·mL^-1. In vitro cell experiments, the killing effect of LEHP was about 1.5 times that of LEP, about 1.2 times that of LEP, and ROS production was about 1.8 times that of LEP. 【Conclusion】 The preparation conditions of LEHP was optimized, and cell experiments showed that LEHP can promote tumor cell apoptosis by improving hypoxia and increasing ROS production, which is expected to provide a safe and effective new method for drug resistance caused by tumor hypoxia.

OBJECTIVES: Safe and effective anticoagulation is essential for hemodialysis patients who are at high risk of bleeding. The purpose of this trial is to evaluate the effectiveness and safety of two-stage regional citrate anticoagulation (RCA) combined with sequential anticoagulation and standard calcium-containing dialysate in intermittent hemodialysis (IHD) treatment. METHODS: Patients at high risk of bleeding who underwent IHD from September 2019 to May 2021 were prospectively enrolled in 13 blood purification centers of nephrology departments, and were randomly divided into RCA group and saline flushing group. In the RCA group, 0.04 g/mL sodium citrate was infused from the start of the dialysis line during blood draining and at the venous expansion chamber. The sodium citrate was stopped after 3 h of dialysis, which was changed to sequential dialysis without anticoagulant. The hazard ratios for coagulation were according to baseline. RESULTS: A total of 159 patients and 208 sessions were enrolled, including RCA group (80 patients, 110 sessions) and saline flushing group (79 patients, 98 sessions). The incidence of severe coagulation events of extracorporeal circulation in the RCA group was significantly lower than that in the saline flushing group (3.64% vs. 20.41%, P<0.001). The survival time of the filter pipeline in the RCA group was significantly longer than that in the saline flushing group ((238.34±9.33) min vs. (221.73±34.10) min, P<0.001). The urea clearance index (Kt/V) in the RCA group was similar to that in the saline flushing group with no statistically significant difference (1.12±0.34 vs. 1.08±0.34, P=0.41). CONCLUSIONS Compared with saline flushing, the two-stage RCA combined with a sequential anticoagulation strategy significantly reduced extracorporeal circulation clotting events and prolonged the dialysis time without serious adverse events.
Humans ; Citric Acid/adverse effects* ; Prospective Studies ; Sodium Citrate ; Hemorrhage/chemically induced* ; Citrates/adverse effects* ; Anticoagulants/adverse effects* ; Renal Dialysis/adverse effects*

Contrast induced nephropathy (CIN) is acute renal injury following administration of contrast media during angiographic or other medical procedures, which represents as the third cause of hospital-acquired renal failure. CIN is associated with prolonged hospital stay, increased health-care costs, and undesirable clinical outcome. The risk of CIN includes advanced age and diabetes mellitus. With the rapid development of iconography and the wide application of interventional techniques, the patients with CIN are increasing. The preventive measures of CIN include hydration, using appropriate contrast media, stopping nephrotoxic drugs, ischemic preconditioning, renal replacement therapy, and using appropriate drugs. In this paper, the current status and early prevention progress of CIN will be reviewed from three aspects of the high-risk factors, pathogenesis and prevention, aiming to provide guidance for the early prevention of CIN and explore new research directions.