Cas13b-mediated RNA targeted therapy alleviates genetic dilated cardiomyopathy in mice.

BACKGROUND: Recent advances in gene editing technology have opened up new avenues for in vivo gene therapy, which holds great promise as a potential treatment method for dilated cardiomyopathy (DCM). The CRISPR-Cas13 system has been shown to be an effective tool for knocking down RNA expression in mammalian cells. PspCas13b, a type VI-B effector that can be packed into adeno-associated viruses and improve RNA knockdown efficiency, is a potential treatment for diseases characterized by abnormal gene expression. RESULTS: Using PspCas13b, we were able to efficiently and specifically knockdown the mutant transcripts in the AC16 cell line carrying the heterozygous human TNNT2R141W (hTNNT2R141W) mutation. We used adeno-associated virus vector serotype 9 to deliver PspCas13b with specific single guide RNA into the hTNNT2R141W transgenic DCM mouse model, effectively knocking down hTNNT2R141W transcript expression. PspCas13b-mediated knockdown significantly increased myofilament sensitivity to Ca2+, improved cardiac function, and reduced myocardial fibrosis in hTNNT2R141W DCM mice. CONCLUSIONS: These findings suggest that targeting genes through Cas13b is a promising approach for in vivo gene therapy for genetic diseases caused by aberrant gene expression. Our study provides further evidence of Cas13b's application in genetic disease therapy and paves the way for future applicability of genetic therapies for cardiomyopathy.

Construction of recombinant adenovirus vector of Uhrf1 gene and its role in DNA damage repair of cardiomyocytes / 中国病理生理杂志

AIM:To construct a recombinant adenovirus vector carrying the mouse ubiquitin-like with plant homeodomain and RING finger domains 1(Uhrf1)gene,validate the expression of Uhrf1 in neonatal mouse cardiomyo-cytes and explored its role in hydrogen peroxide(H2O2)-induced DNA damage.METHODS:The mouse Uhrf1 gene cod-ing sequence was amplified by polymerase chain reaction(PCR),digested,and inserted into the pADM-CMV-C-FH vec-tor to create the recombinant adenoviral plasmid ADM-Uhrf1.Following transfection into HEK293T cells,we generated re-combinant adenoviral particles,amplified,purified,and determined the titer.Neonatal mouse cardiomyocytes were infect-ed at an multiplicity of infection(MOI)of 50,UHRF1 protein expression was validated via Western blot and immunofluo-rescence staining.H2O2-induced DNA damage was explored along with adenovirus-mediated Uhrf1 overexpression to inves-tigate its role in DNA damage repair.RESULTS:ADM-Uhrf1 virus titer,determined by capsid immunofluorescence as-say,was 1.8×1013 pfu/L.Western blot confirmed a significant increase in UHRF1 protein expression(P＜0.05),with im-munofluorescence indicating predominant nuclear localization.Uhrf1 overexpression effectively inhibited the expression of the DNA damage marker,phosphorylated H2AX protein(γH2AX)(P＜0.01).CONCLUSION:We successfully con-structed a recombinant adenoviral vector carrying the mouse Uhrf1 gene,facilitating Uhrf1 overexpression in neonatal mouse cardiomyocytes.Furthermore,this overexpression effectively alleviated DNA damage in cardiomyocytes.

Technical points of modified (port-free) single-site robot-assisted laparoscopic radical prostatectomy / 现代泌尿外科杂志

With the development of minimally invasive technology, robot-assisted laparoscopic radical prostatectomy (RARP) has become the main method and gold standard in the treatment of organ-localized prostate cancer. After previous exploration of various surgical approaches and surgical methods in our center, we first proposed the modified (port-free) single-site RARP (pf-ssRARP), which has been proved safe and feasible by theoretical verification and practical operation. The technique has certain advantages in postoperative rehabilitation, urinary control recovery, sexual function improvement, incision cosmetics and social economics. In this paper, the key steps of this technique are introduced and illustrated in detail.

Mucus-penetrating nonviral gene vaccine processed in the epithelium for inducing advanced vaginal mucosal immune responses

Establishment of vaginal immune defenses at the mucosal interface layer through gene vaccines promise to prevent infectious diseases among females. Mucosal barriers composed of a flowing mucus hydrogel and tightly conjugated epithelial cells (ECs), which represent the main technical difficulties for vaccine development, reside in the harsh, acidic human vaginal environment. Different from frequently employed viral vectors, two types of nonviral nanocarriers were designed to concurrently overcome the barriers and induce immune responses. Differing design concepts include the charge-reversal property (DRLS) to mimic a virus that uses any cells as factories, as well as the addition of a hyaluronic acid coating (HA/RLS) to directly target dendritic cells (DCs). With a suitable size and electrostatic neutrality, these two nanoparticles penetrate a mucus hydrogel with similar diffusivity. The DRLS system expressed a higher level of the carried human papillomavirus type 16 L1 gene compared to HA/RLS in vivo. Therefore it induced more robust mucosal, cellular, and humoral immune responses. Moreover, the DLRS applied to intravaginal immunization induced high IgA levels compared with intramuscularly injected DNA (naked), indicating timely protection against pathogens at the mucus layer. These findings also offer important approaches for the design and fabrication of nonviral gene vaccines in other mucosal systems.

Short term follow-up of unstable femoral neck fractures in young adults treated with femoral neck system and cannulated compression screws / 中国骨伤

OBJECTIVE@#To investigate the clinical efficacy of femoral neck system(FNS) and three cannulated compression screws(CCS) in the treatment of unstable femoral neck fractures in young adults.@*METHODS@#The clinical data of 52 young and middle-aged patients with unstable femoral neck fractures admitted from August 2018 to August 2021 were reviewed and analyzed. All patients were divided into two groups according to the internal fixation method, 25 cases were treated with FNS fixation, 27 cases were treated with closed reduction and 3 CCS inverted triangular distribution. The operation time, incision length, intraoperative bleeding, hospitalization expenses and fracture reduction quality of two groups were recorded and compared;The patients were followed up regularly after operation. The fracture healing time, complete weight-bearing time and postoperative complications(nonunion, femoral neck shortening, femoral head necrosis) of two groups were compared. The Harris score was used to evaluate hip function 6 months after surgery.@*RESULTS@#The operation was successfully completed in both groups. The patients in FNS group had more bleeding, longer incision length and higher hospitalization cost than CCS group(P<0.01). There ware no significant difference in operation time and Garden index between two groups(P>0.05). Patients in both groups were followed up for 6 to 32 months.The fracture healing time in FNS group was less than that in CCS group, the time of complete weight bearing after surgery was earlier than that in CCS group, and the hip Harris score was higher than that in CCS group (P<0.01). There were no internal fixation fracture complications in two groups during follow-up. In the FNS group, there were 4 cases of avascular necrosis of the femoral head and 2 cases of femoral neck shortening, of which 3 cases underwent total hip replacement due to avascular necrosis of the femoral head. In the CCS group, there were 2 cases of nonunion, 9 cases of avascular necrosis of the femoral head, and 11 cases of femoral neck shortening, among which 5 cases underwent total hip replacement due to nonunion and avascular necrosis of the femoral head.@*CONCLUSION@#With simple operation, rotational stability and angular stability, FNS enables patients to start functional exercise as early as possible and reduces the incidence of postoperative complications of unstable femoral neck fracture. It is a new choice for the treatment of unstable femoral neck fracture in young adults.

Lung-protective effect of pressure-controlled volume-guaranteed ventilation combined with dexmedetomidine in pediatric patients undergoing laparoscopic surgery / 中华麻醉学杂志

Objective:To evaluate the protective effect of pressure-controlled volume-guaranteed ventilation (PC-VG) combined with dexmedetomidine on the lung of pediatric patients undergoing laparoscopic surgery.Methods:Forty-eight pediatric patients of either sex, aged 2-6 yr, weighing 8-21 kg, scheduled for elective laparoscopic pyeloplasty, were divided into 3 groups ( n=16 each) using a random number table method: volume-controlled ventilation (VCV) group (V group), PC-VG group (P group), and PC-VG combined with dexmedetomidine group (PD group). In PD group, dexmedetomidine was intravenously infused for 15 min at a loading dose of 0.5 μg/kg starting from the time point before anesthesia induction followed by a continuous infusion of 0.2-0.5 μg·kg -1·h -1 until the end of operation. VCV mode was used in group V, and PC-VG mode was used in P and PD groups, ventilator settings were adjusted to the mode with a tidal volume 6-8 ml/kg, respiratory rate 15-25 breaths/min, inspiratory/expiratory ratio 1∶2, oxygen flow rate 2 L/min, fraction of inspired oxygen 60%, and P ETCO 2 was maintained at 35-40 mmHg during mechanical ventilation in three groups. At 5 min before pneumoperitoneum (T 0), 10, 60 and 120 min of pneumoperitoneum (T 1-3) and 10 min after release of pneumoperitoneum pressure (T 4), peak airway pressure (Ppeak), mean airway pressure (Pmean), compliance of lung (C L) and airway resistance (Raw) were recorded, alveolar-arterial oxygen partial pressure difference (PA-aO 2), oxygenation index (OI), and respiratory index (RI) were recorded. The occurrence of pulmonary complications was recorded within 7 days after operation. Results:Compared with V group, the Ppeak and Raw were significantly decreased and C L was increased at T 1-4, and PA-aO 2 and RI were decreased and OI was increased at T 3, 4 in P group and PD group ( P<0.05). Compared with P group, no significant change was found in the parameters of respiratory mechanics mentioned above at each time point ( P>0.05), and PA-aO 2 and RI were decreased and OI was increased at T 3, 4 in PD group ( P<0.05). There was no significant difference in the incidence of postoperative pulmonary complications among the three groups ( P>0.05). Conclusions:PC-VG combined with dexmedetomidine has a certain lung-protective effect in pediatric patients undergoing laparoscopic surgery.

Study of senescence protein p66Shc on myocardial tissue repair in adult mice / 生理学报

Our previous study has shown that p66Shc plays an important role in the process of myocardial regeneration in newborn mice, and p66Shc deficiency leads to weakened myocardial regeneration in newborn mice. This study aims to explore the role of p66Shc protein in myocardial injury repair after myocardial infarction in adult mice, in order to provide a new target for the treatment of myocardial injury after myocardial infarction. Mouse myocardial infarction models of adult wild-type (WT) and p66Shc knockout (KO) were constructed by anterior descending branch ligation. The survival rate and heart-to-body weight ratio of two models were compared and analyzed. Masson's staining was used to identify scar area of injured myocardial tissue, and myocyte area was determined by wheat germ agglutinin (WGA) staining. TUNEL staining was used to detect the cardiomyocyte apoptosis. The protein expression of brain natriuretic peptide (BNP), a common marker of myocardial hypertrophy, was detected by Western blotting. The results showed that there was no significant difference in survival rate, myocardial scar area, myocyte apoptosis, and heart weight to body weight ratio between the WT and p66ShcKO mice after myocardial infarction surgery. Whereas the protein expression level of BNP in the p66ShcKO mice was significantly down-regulated compared with that in the WT mice. These results suggest that, unlike in neonatal mice, the deletion of p66Shc has no significant effect on myocardial injury repair after myocardial infarction in adult mice.

The impacts of corrected left ventricular ejection time combined with dobutamine on hepatocellular carcinoma patients.

To evaluate the clinical effect of corrected left ventricular ejection time (LVETc) combined with dobutamine on the intraoperative management of patients undergoing hepatectomy for hepatocellular carcinoma. Sixty-eight patients with elective proposed pancreaticoduodenectomy, aged 61-78 years, body mass index 19-26 kg/m2, and ASA classification II or III, were divided into two groups (n = 34) using the random number table method: the esophageal ultrasound group (S group) and the esophageal ultrasound combined with dobutamine group (D group). In both groups, an esophageal ultrasound probe was placed after induction of anesthesia, and the left ventricular ejection time (LVET) and stroke volume (SV) were measured via a long-axis section of gastric fundus to guide fluid infusion. Nitroglycerin or a combination of dobutamine and nitroglycerine were pumped intravenously from the beginning of surgery to the completion of hemostasis after partial hepatectomy, in groups S or D, respectively. Central Venous Pressure (CVP), heart rate HR, and mean arterial pressure MAP were recorded at entry (T0), immediately after induction (T1), at the beginning of the operation (T2), during hilar occlusion (T3), after partial hepatectomy (T4), and after the operation (T5). SV and LVETc were recorded between T1 and T5. At T0 and T5, blood samples from radial artery and central vein were taken to determine the concentration of blood lactic acid, and the oxygen supply index (DO2I) and oxygen uptake rate (O2ERe) were calculated by blood gas analysis. The operation time, hilar occlusion time, intraoperative urine volume, intraoperative crystalloid and colloid infusion, intraoperative blood loss and blood transfusion, and the occurrence of cardiac gas emboli during the operation were also recorded. Adverse events of cardiovascular, pulmonary, and renal function during and after operation were registered. Sixty-four patients were included in the final analysis. Compared with group S, group D had lower CVP values at T2-T3 and higher SV values at T2-T5, reduced intraoperative blood loss, significantly increased intraoperative urine output, a smaller total dose of nitroglycerin use, and lower incidences of intraoperative hypotension and cardiac gas emboli (P < 0.05). Esophageal ultrasound detection of LVETc combined with dobutamine ensures hemodynamic stability in patients undergoing partial hepatectomy while reducing the incidence of intraoperative hypotension and air embolism.

Clinical application effects of two longitudes three transverses method in perforator location of thoracodorsal artery perforator flap and deep wound repair / 中华烧伤杂志

Objective: To explore the clinical application value of two longitudes three transverses method in the location of the perforator of thoracodorsal artery perforator and deep wound repair. Methods: The retrospectively observational study was conducted. From December 2018 to June 2020, 17 patients with deep wounds who were admitted to the Affiliated Hospital of Zunyi Medical University met the inclusion criteria and were included in this study, including 7 males and 10 females, aged 12 to 72 years. The wound areas of patients after debridement were 7 cm×3 cm to 11 cm×7 cm. Two longitudinal lines were located through the midpoint of the armpit, the posterior superior iliac spine, and the protruding point of the sacroiliac joint, and three transverse lines were located 5, 10, and 15 cm below the midpoint of the armpit between the two longitudinal lines, i.e. two longitudes three transverses method, resulting in two trapezoidal areas. And then the thoracodorsal artery perforators in two trapezoidal areas were explored by the portable Doppler blood flow detector. On this account, a single or lobulated free thoracodorsal artery perforator flap or flap that carrying partial latissimus dorsi muscle, with an area of 7 cm×4 cm to 12 cm×8 cm was designed and harvested to repair the wound. The donor sites were all closed by suturing directly. The number and location of thoracodorsal artery perforators, and the distance from the position where the first perforator (the perforator closest to the axillary apex) exits the muscle to the lateral border of the latissimus dorsi in preoperative localization and intraoperative exploration, the diameter of thoracodorsal artery perforator measured during operation, and the flap types were recorded. The survivals of flaps and appearances of donor sites were followed up. Results: The number and location of thoracodorsal artery perforators located before operation in each patient were consistent with the results of intraoperative exploration. A total of 42 perforators were found in two trapezoidal areas, with 2 or 3 perforators each patient. The perforators were all located in two trapezoid areas, and a stable perforator (the first perforator) was located and detected in the first trapezoidal area. There were averagely 1.47 perforators in the second trapezoidal area. The position where the first perforator exits the muscle was 2.1-3.1 cm away from the lateral border of the latissimus dorsi. The diameters of thoracodorsal artery perforators were 0.4-0.6 mm. In this group, 12 cases were repaired with single thoracodorsal artery perforator flap, 3 cases with lobulated thoracodorsal artery perforator flap, and 2 cases with thoracodorsal artery perforator flap carrying partial latissimus dorsi muscle. The patients were followed up for 6 to 16 months. All the 17 flaps survived with good elasticity, blood circulation, and soft texture. Only linear scar was left in the donor area. Conclusions: The two longitudes three transverses method is helpful to locate the perforator of thoracodorsal artery perforator flap. The method is simple and reliable. The thoracodorsal artery perforator flap designed and harvested based on this method has good clinical effects in repairing deep wound, with minimal donor site damage.

Extraperitoneal robot-assisted laparoscopic radical prostatectomy through single incision: Establishment and application of a modified channel / 中华男科学杂志

Objective@#To assess the feasibility and validity of the establishment of a modified channel for extraperitoneal robot-assisted laparoscopic radical prostatectomy (RARP) through single incision.@*METHODS@#From November 2020 to January 2021, 35 cases of localized PCa were treated by extraperitoneal RARP through single incision in our center. All the operations were performed by the same surgeon, none via the multichannel port for the establishment of the channel. We recorded and analyzed the intra- and postoperative parameters, operation cost, complications, pathological findings and follow-up data.@*RESULTS@#All the operations were successfully completed, without conversion to open surgery or additional channels, or serious postoperative complications, the time for establishing the extraperitoneal space averaging 25.4 (20.0－45.0) min, the operation time 67.3 (35.0－125.0) min, intraoperative blood loss 75.5 (60.0－150.0) ml, time to first postoperative anal exhaust 26 (8－48) h, and postoperative hospital stay 7.89 (7－10) d. Postoperative pathology showed adenocarcinoma in all the cases, with Gleason score (GS) 3+3 in 9 (25.7%), GS 3+4 in 9 (25.7%), GS 4+3 in 8 (22.9%), and GS ≥ 8 in 9 (25.7%) of the cases, 23 (65.7%) in the

Comparison of clinical efficacy of transperitoneal robot assisted laparoscopic radical prostatectomy versus extraperitoneal single port robot assisted laparoscopic radical prostatectomy / 中华泌尿外科杂志

Objective:To compare the clinical efficacy of transperitoneal robot assisted laparoscopic radical prostatectomy (RARP) versus extraperitoneal single port RARP.Methods:A retrospective analysis was perfoumed on 142 cases of RARP from July 2019 to June 2020 in Robotic Minimally Invasive Center of Sichuan Provincial People's Hospital, including 82 cases(Group A) , aged 70.0(65.6-78.0), undergoing transperitoneal RARP and 60 cases(Group B), aged 68.2 years old(60.1-79.2), undergoing extraperitoneal single port RARP. For group A, PSA was 12.9(5.6-64.0) ng/ml, with 26 cases of less than 10 ng/ml(31.7%), 40 cases of 10-20 ng/ml(48.8%), and 16 cases of more than 20 ng/ml(19.5%), the Gleason score was 7.2(6.0-10.0), with 14 cases(17.1%) of ≤6, 56 cases(68.3%) of 7, and 12 cases(14.6%) of ≥8, and the prostate volume was 61.3(29.0-112.0) ml. There were 49 cases with BMI≤25 kg/m 2, accounting for 59.8%, and 33 cases with BMI>25 kg/m 2, accounting for 40.2%. There were 17 cases(20.7%) of T 1, 44 cases(53.7%) of T 2 and 21 cases(25.6%) of T 3. The proportion of lymph node dissection was 17.1%, and 4 cases(4.9%)had a history of operation. For group B, the PSA was 12.2(1.0-42.6)ng/ml, with 20 cases (33.3%) of <10 ng/ml, 31 cases(51.7%)of 10-20 ng/ml, and 9 cases (15%) of >20 ng/ml. Gleason score was 7.1(6.0-9.0), with 12 cases (20.0%) of ≤6, 42 cases (70.0%) of 7, and 6 cases (10.0%)of ≥8. Prostate volume was 42.4(31.2-72.8)ml on average. There were 37 cases (61.7%) with BMI≤25 kg/m 2, and 23 cases (38.3%)with BMI >25 kg/m 2 . There were 17 cases(28.3%)of T 1, 32 cases(53.3%)of T 2 and 11 cases(18.3%)of T 3.The proportion of lymph node dissection was 11.7% and 4 cases (6.7%) had a history of operation.There was no statistically significant difference in term of age, PSA level, Gleason score, BMI, clinical stage, proportion of lymph node dissection or history of operation between the two groups( P>0.05), but there was statistically significant difference for prostate volume( P<0.05). All operations were performed by the same operator. Four different ways of bladder neck and urethral dissociation was selected according to the intraoperative conditions in Group A, include VIP style, T-shape incision style, VIP plus T-shape incision style or the style along the lateral side of the bladder neck. Small and single anterograde incision stripping of bladder neck was routinely performed in the Group B. Postoperative follow-up was performed to compare the operation time, intraoperative blood loss, bladder neck and urethral anastomosis time, postoperative hospital stay, postoperative exhaust time, postoperative complications, positive rate of surgical margin, indwelling time of urinary catheter, urinary continence satisfaction rate of immediately after operation, 3 months and 6 months after operation, wound healing and aesthetics. Results:All of the operations were successfully completed under robot-assisted laparoscopy, and there was no conversion to open surgery. The operation time was 56.0(45.0-112.0) min in the Group A and 65.4(55.5-96.8) min in the Group B, and there was no statistically significant difference( P>0.05). The intraoperative blood loss was 76.2(30.0-120.5) ml and 55.6(45.5-114.6) ml, respectively, and the difference was not statistically significant( P>0.05). The time of bladder neck urethral anastomosis was 18.9(12.6-25.6) min and 16.2(10.7-19.3) min, respectively, and the difference was not statistically significant( P>0.05). The postoperative hospital stay days were 9.3(8.0-16.0) d and 8.4(7.0-13.0) d, respectively, and the difference was not statistically significant( P>0.05). The postoperative exhaust time was 1.3(0.7-3.0) d and 3.4(2.0-7.0) d, respectively, and the difference was statistically significant( P<0.05). There was 1 case of anastomotic fistula with ureteral injury in Group A, and no serious complication in Group B, and the difference was not statistically significant( P>0.05). The number of positive surgical margin in the two groups was 13(15.9%)and 9(15.0%)respectively, and the difference was not statistically significant( P>0.05). The indwelling time of urinary catheter after operation was 9(7-21) d and 6(4-8) d, respectively, and the difference was statistically significant( P<0.05). The number of patients with satisfactory urinary continence immediately after surgery, 3 months and 6 months after surgery in the two groups were 8(9.8%), 51(62.2%), 62(75.6%) and 17(28.3%), 43(71.7%) and 54(90.0%), respectively. The differences were statistically significant( P<0.05). The total incision lengths in the two groups were 12.1(10.4-13.4) cm and 5.6(5.0-6.0) cm, respectively, and the difference was statistically significant( P<0.05). Conclusions:The extraperitoneal single port RARP is safe and feasible, and the postoperative effect is similar to that of transperitoneal RARP. It has the advantages of shorter recovery time, higher urinary continence satisfaction rate, neater and more beautiful incision. The long-term therapeutic effect needs further confirming by prospective study.

The modified single incision robot-assisted laparoscopic radical prostatectomy: initial experience and clinical efficiency / 中华泌尿外科杂志

Objective:To evaluate the feasibility and clinical efficiency of robot-assisted laparoscopic radical prostatectomy (RARP) via extraperitoneal PORT-free single incision approach.Methods:The data of 33 patients with prostate cancer underwent the extraperitoneal PORT-free single incision RARP from November 2020 to January 2021 in Sichuan Provincial People's Hospital was retrospectively reviewed. The average age was 66.7 (58-78) years, the median PSA was 20.77 (2.89, 56.44) ng/m, and the mean Gleason score was 7.0 (6.0-9.0). The mean prostate volume was 48.4 (25.0-220.0) ml. Clinical stage: 32 cases was in cT 2a-2cN 0M 0, 1 case in cT 3aN 0M 0. 16 cases had a history of operation. All 33 operations were performed by the same operator. All operations were performed by extraperitoneal PORT-free single-incision approach. The surgical condition, postoperative complication, pathology, and follow-up results were observed. Results:In this study, 33 operations were successfully completed without conversion to open or additional single hole channel instruments. The average operation time was 61.3 (38.0-120.0) min, with the mean intraoperative bleeding volume of 72.2 (45.0-220.0) ml and the mean bladder neck urethral anastomosis time of 11.7 (8.5-15.7) min. The mean postoperative hospital stay was 7.9 (6.0-15.0) d, the mean postoperative indwelling time of urinary catheter was 6.8 (6.0-14.0) d, and the mean postoperative evacuation time was 1.0 (0.5-3.0) d. The average incision length was 5.2 (4.6-5.8) cm. There was no obvious complications. The postoperative pathological stage: 21 cases were in < pT 3a, 12 cases were in ≥ pT 3a, and 6 cases (18.8%) had positive resection margin. 29 cases (88.9%) acquired satisfactory urinary continence after operation, and the frequency of urinary pad use was ≤ 1 tablet/day. Conclusions:The extraperitoneal single-incision RARP surgical channel without PORT is safe and feasible with a satisfying cosmetic effect, which saves costs and requires less specific channel device. Simultaneously, the new approach has strong replicability, short-term tumor control and urinary control effect with rapid postoperative recovery. However, the sample size of this study is relatively small, which needs further research and demonstration

A Novel Long-circulating DOX Liposome: Formulation and Pharmacokinetics Studies.

BACKGROUND: Doxorubicin (DOX) is a leading chemotherapeutic in cancer treatment because of its high potency and broad spectrum. Liposomal doxorubicin (Doxil®) is the first FDA-approved PEG-liposomes of DOX for the treatment of over 600,000 cancer patients, and it can overcome doxorubicin-induced cardiomyopathy and other side effects and prolong life span. The addition of MPEG2000-DSPE could elevate the total cost of cancer treatment. OBJECTIVE: We intended to prepare a novel DOX liposome that was prepared with inexpensive materials egg yolk lecithin and Kolliphor HS15, thus allowing it to be much cheaper for clinical application. METHODS: DOX liposomes were prepared using the combination of thin-film dispersion ultrasonic method and ammonium sulfate gradient method and the factors that influenced formulation quality were optimized. After formulation, particle size, entrapment efficiency, drug loading, stability, and pharmacokinetics were determined. RESULTS: DOX liposomes were near-spherical morphology with the average size of 90 nm and polydispersity index (PDI) of less than 0.30. The drug loading was up to 7.5%, and the entrapment efficiency was over 80%. The pharmacokinetic studies showed that free DOX could be easily removed and the blood concentration of free DOX group was significantly lower than that of DOX liposomes, which indicated that the novel DOX liposome had a certain sustainedrelease effect. CONCLUSION: In summary, DOX liposome is economical and easy-prepared with prolonged circulation time. Lay Summary: Doxorubicin (DOX) is a leading chemotherapeutic in cancer treatment because of its high potency and broad spectrum. Liposomal doxorubicin (Doxil®) is the first FDAapproved PEG-liposomes of DOX to treat over 600.000 cancer patients, overcoming doxorubicin- induced cardiomyopathy and other side effects and prolonging life span. The addition of MPEG2000-DSPE could elevate the total cost of cancer treatment. We intend to prepare a novel DOX liposome prepared with inexpensive materials egg yolk lecithin and Kolliphor HS15, thus allowing it to be much cheaper for clinical use. The novel DOX liposome is economical and easy-prepared with prolonged circulation time.

Clinical application of different bladder neck separation techniques in robot-assisted laparoscopic radical prostatectomy / 中华泌尿外科杂志

Objective:To investigate the clinical effect of different bladder neck separation methods in robot-assisted laparoscopic radical prostatectomy (RARP).Methods:To retrospective analysis the data of robot-assisted laparoscopic radical prostatectomy (RARP)in our center from October, 2014 to October, 2018. All operations were performed by the same urologist. According to the different methods of bladder neck separationAccording to the different methods of bladder neck separation, all the patients were divided into four groups. Group A routine forward peeling method (500 cases): Make a 1cm incision at 12 o'clock on the front of the bladder neck, cut off the detrusor muscle and cut the bladder neck. Group B T-cut incision of the bladder neck (133 cases): identify the bladder and prostate Junction, T-shaped incision of the anterior wall of the bladder neck. Group C conventional stripping method combined with T-shaped incision of the bladder neck (81 cases). Group D lateral approach (36 cases): along the lateral side of the bladder neck and the medial posterior ligament of the prostate is separated and merges with the previously established Dirichlet space. The general data of patients were analyzed statistically.The average ages of groups A, B, C, and D were 63 years (62.5 to 67 years), 65 years (61 to 68 years), 66 years (64.5 to 70.5 years), and 62 years (59.5 to 66.5 years)respectively, there was no statistical significance difference in terms of age in 4 groups( P>0.05); PSA is 13 ng/ml(9.0 to 22 ng/ml), 7.4 ng/ml(6.4 to 26.0 ng/ml), 6.2 ng/ml(5.3 to 27.0 ng/ml), 14ng/ml(8.4 to 21.0 ng/ml), ( P>0.05); Gleason scores of puncture were 6.9(5 to 9), 7(6 to 12), 9(8 to 16), 10(6 to 18), ( P>0.05); the prostate volume was 66ml(42 to 78 ml), 70ml(50 to 89 ml), 53ml (43 to 72 ml), 80 ml (68 to 92 ml), ( P>0.05); the proportions of body mass index ≤25 kg/m 2 were 60.0%, 63.9%, 39.1%, 42.0%, and>25 kg/m 2 were 40%, 36.1%, 60.9%, and 58.0%, respectively, ( P>0.05). The operation time, bleeding volume, anastomosis time, postoperative hospital stay, postoperative complications, positive rate of proximal incision margin, urinary indwelling time, and urinary control rate in the four groups analyzed. Results:All 750 RARP operations were successful, and none were converted to open.The operation time of groups A, B, C, and D were 100 min(70 to 120 min), 89 min(70 to 95 min), 105 min(80 to 127 min), and 110 min(90 to 130 min), ( P>0.05); anastomosis time was 20.5 min (18.0 to 25.0 min)、16.1min (10.7 to 17.3 min)、25.4 min (18.9 to 27.0 min)、and 28.5 min (21.0 to 32.0 min), the anastomosis time in group B was significantly shorter than other groups ( P<0.05); the postoperative hospital stays were 9.3 days (8.0 to 13.0 days), 8.4 days (6.0 to 16.0 days), 10.8 days (8.0 to 16.0 days)and 7.8 days (7.0 to 14.0 days), ( P>0.05). Postoperative complications: Anastomotic fistula and ureteral injury occurred in 3 cases in group A, and no serious complications occurred in the other 3 groups. Proximal marginal positive rate: 40 cases (8.0%) in group A, 0 cases in group B, 6 cases (7.3%) in group C, 3 cases (8.3%) in group D, and low positive rate of margin incision in group B( P<0.05). The urinary indwelling time was 7 d (6 to 8 d), 6 d(4 to 8 d), 12 d(6 to 18 d), 10 d(6 to 13 d), ( P>0.05). Six-month postoperative urine control rate: 381 cases (75.2%) in group A, 102 cases (76.9%) in group B, 61 (75.4%) in group C, and 27 (73.8%) in group D, ( P>0.05). Conclusions:The above four method of bladder neck separation during robot-assisted laparoscopic radical prostatectomy is safe and feasible, which can effectively avoid ureteral damage. Each method can obtain better urine control within six months after surgery rate. The positive rate of proximal incision margin after T-shaped bladder neck was lowest among four groups.

The experience of suprapubic extraperitoneal single-port robot-assisted radical prostatectomy / 中华泌尿外科杂志

Six patients with prostate cancer, treated by suprapubic extraperitoneal single-port robot-assisted radical prostatectomy, had been studied retrospectively from December 2019 to January 2020. All 6 patients have been treated by suprapubic peritoneum single port robot assisted laparoscopic surgery without other channels. The robot assisted laparoscopic radical prostatectomy via suprapubic peritoneum is safe and feasible when based on reasonable selection criteria of patients. It has been shown that the postoperative recovery was fast and the tumor control and continence rate were good under the short-term follow-up. However, the long-term outcome should be evaluated by a long-term follow-up.

Erratum to: Exosome-derived galectin-9 may be a novel predictor of rejection and prognosis after liver transplantation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Erratum to: J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2019 20(7):605-612. https://doi.org/10.1631/jzus.B1900051. The original version of this article unfortunately contained a mistake. In p.605, the number of the Zhejiang Provincial Natural Science Foundation of China (No. Y17H160118) in Funding is incorrect. The correct number should be LY17H160026, which is the approval number of the project, whereas Y17H160118 is the application number of the project.

Exosome-derived galectin-9 may be a novel predictor of rejection and prognosis after liver transplantation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Acute cellular rejection (ACR) remains a major concern after liver transplantation. Predicting and monitoring acute rejection by non-invasive methods are very important for guiding the use of immunosuppressive drugs. Many studies have shown that exosomes and their contents are potential biomarkers for various liver diseases. Here, we identify and validate the role of exosomes and galectin-9 in ACR after liver transplantation. Exosomes were isolated from three sets of paired patients, with and without ACR, and the proteins within the exosomes were isolated and identified. Candidate proteins were then validated using a tissue microarray containing resected liver samples from 73 ACR and 63 non-rejection patients. Finally, protein expression and clinical manifestations were included in Kaplan-Meier survival and Cox regression analyses. Circulating exosomes were isolated from ACR and non-rejection patients and characterized using transmission electron microscopy and western blotting for CD63/CD81. Western blotting experiments revealed higher levels of galectin-9 protein in circulating exosomes from ACR recipients. Immunohistochemical analysis of the tissue microarray showed that the expression of galectin-9 in resected liver was significantly higher in the ACR group than in the non-rejection group (P<0.05). Higher levels of galectin-9 expression in resected livers were associated with poorer prognosis (P<0.05). Exosome-derived galectin-9 may be a novel predictor of rejection and prognosis after liver transplantation.

Comparison of different approaches to fascia iliaca compartment block for postoperative analgesia in elderly patients undergoing total hip arthroplasty / 中华麻醉学杂志

Objective To compare the efficacy of ultrasound-guided fascia iliaca compartment block(FICB)above inguinal ligament and longitudinal inguinal FICB for postoperative analgesia in elderly patients undergoing total hip arthroplasty(THA).Methods Sixty American Society of Anesthesiologists physical statusⅠ orⅡ patients of both sexes,aged 65-76 yr,with body mass index of 19-26 kg/m2,scheduled for elective unilateral THA,were divided into 2 groups(n＝30 each)using a random number ta-ble method: FICB above inguinal ligament group(S group)or longitudinal inguinal FICB group(G group).After the end of surgery,patients received ultrasound-guided FICB using a short-axis in-plane technique in S group and longitudinal inguinal FICB using a long-axis in-plane technique in G group,and patients received 0.4％ropivacaine as a loading dose of 40 ml,followed by continuous infusion of 0.2％ropivacaine 5 ml/h for 48 h.Ultrasound imaging time,puncture injection time and operating time of FICB were recorded.The efficacy of nerve block,effective pressing times,cumulative consumption of ropiva-caine,satisfaction with analgesia,and development of related complications were recorded at 6,12,24,36,48 and 72 h after surgery(T1-6).Results Compared with group G,the requirement for rescue anal-gesia with dezocine was significantly decreased,the effective pressing times at T3-5 and cumulative consump-tion of ropivacaine at T3,4 were reduced,and the success rate of obturator nerve block was increased at T1-6 in group S(P＜0.05).There were no significant differences in the success rate of the femoral nerve and lat-eral femoral cutaneous nerve block,satisfaction with analgesia and development of related complications be-tween the two groups(P>0.05).Conclusion Ultrasound-guided FICB above inguinal ligament increases the success rate of the obturator nerve block,provides better efficacy for postoperative analgesia and is more helpful for early postoperative functional exercise and recovery than the longitudinal inguinal FICB in the pa-tients undergoing THA.

Comparison of different approaches to fascia iliaca compartment block for postoperative analgesia in elderly patients undergoing total hip arthroplasty / 中华麻醉学杂志

Objective@#To compare the efficacy of ultrasound-guided fascia iliaca compartment block(FICB) above inguinal ligament and longitudinal inguinal FICB for postoperative analgesia in elderly patients undergoing total hip arthroplasty (THA).@*Methods@#Sixty American Society of Anesthesiologists physical statusⅠ orⅡ patients of both sexes, aged 65-76 yr, with body mass index of 19-26 kg/m2, scheduled for elective unilateral THA, were divided into 2 groups (n=30 each) using a random number table method: FICB above inguinal ligament group (S group) or longitudinal inguinal FICB group (G group). After the end of surgery, patients received ultrasound-guided FICB using a short-axis in-plane technique in S group and longitudinal inguinal FICB using a long-axis in-plane technique in G group, and patients received 0.4% ropivacaine as a loading dose of 40 ml, followed by continuous infusion of 0.2% ropivacaine 5 ml/h for 48 h. Ultrasound imaging time, puncture injection time and operating time of FICB were recorded.The efficacy of nerve block, effective pressing times, cumulative consumption of ropivacaine, satisfaction with analgesia, and development of related complications were recorded at 6, 12, 24, 36, 48 and 72 h after surgery (T1-6).@*Results@#Compared with group G, the requirement for rescue analgesia with dezocine was significantly decreased, the effective pressing times at T3-5 and cumulative consumption of ropivacaine at T3, 4 were reduced, and the success rate of obturator nerve block was increased at T1-6 in group S (P<0.05). There were no significant differences in the success rate of the femoral nerve and lateral femoral cutaneous nerve block, satisfaction with analgesia and development of related complications between the two groups (P>0.05).@*Conclusion@#Ultrasound-guided FICB above inguinal ligament increases the success rate of the obturator nerve block, provides better efficacy for postoperative analgesia and is more helpful for early postoperative functional exercise and recovery than the longitudinal inguinal FICB in the patients undergoing THA.

Effect of epidermal growth factor receptor signaling pathway inhibitor gefitinib on fracture healing in rats / 中华创伤杂志

Objective To observe the effect of epidermal growth factor receptor (EGFR) signal pathway inhibitor gefitinib on femoral fracture healing in rats.Methods Eighty SD rats were divided into experimental group (n =40) and control group (n =40) by random number table.An open femur fractured model was established by cutting the middle part of the femoral shaft and placing 1 mm Kirschner wire through medial and lateral femoral patella incision.Rats in the experimental group were given gefitinib (100 mg · kg-1 · d-1),an inhibitor of epidermal growth factor receptor (EGFR) signaling pathway,and rats in the control group were given equal doses of methylcellulose.The X-ray examination was performed before femoral and serum specimens were collected at 1,2,3,4,and 6 weeks.Bone fracture healing was evaluated by Lane and Sandhu X ray scoring and biomechanical test.Then the tissue morphology was observed by HE staining,safranin fast green staining,and microCT detection.The bone volume (BV),bone volume fracture (BV/TV),trabecular number (Tb.N),trabecular thickness (Tb.Th),and trabecular separation (Tb.Sp) were measured by MicroCT.The levels of serum BALP,PINP,TRACP5b,and CTX were measured by Elisa.Results The X-ray scores of the experimental group (1.26 ±0.54,4.94 ± 1.16,and 8.23 ± 1.17) were significantly higher than those in the control group (0.91 ± 0.52,4.11 ± 1.18,and 7.08 ± 0.86) at 1,2 and 3 weeks after the fracture (P ＜ 0.05).The failure load of experimental group at 3 and 4 weeks after fracture [(38.65 ± 1.07) N,(63.63 ± 7.74)N] were significantly larger than those of the control group [(29.47 + 1.00)N,(45.42 + 3.26) N] (P ＜ 0.05).The callus formation in the experimental group was obvious and the healing process of the fracture was faster than that of the control group according to HE and safranin fast green staining.The results of microCT showed that in the experimental group,BV was significantly higher at 2 and 3 weeks after fracture [(59.30 ± 6.54) mm3 and (43.08 ± 2.33) mm3] than those in control group [(42.39 ± 7.82) mm3 and (33.43 ± 5.94) mm3];BV / TV at 2,3,and 4 weeks after fracture [(0.61 ±0.06)％,(0.55 ±0.05)％,and (0.53 ±0.04)％] were significantly higher than those in the control group [(0.48±0.07)％,(0.44±0.07)％,and (0.43±0.03)％];Tb.N at 2,3,and 4 weeks after fracture [(2.05 ± 0.11)/mm,(1.86 ± 0.18)/mm,and (2.034 ± 0.26)/mm] were significantly higher than those in control group [(1.63 ±0.21)/rmm,(1.32 ±0.21)/rmm,and (1.65 ± 0.08)/mm)];Tb.Th at 2,3,and 4 weeks after fracture [(0.33 ± 0.02) mm,(0.33 ± 0.03) mm,and (0.27 ± 0.02) mm] were significantly higher than those in the control group [(0.27 ±0.03)rmm,(0.28 ± 0.02)mm,and 0.23 ±0.01)mm];Tb.Sp at 3 weeks after fracture [(0.39 ±0.07)mm] was significantly higher than that in the control group [(0.30 ± 0.03) mm] (P ＜ 0.05).The levels of serum BALP in experimental group at 2 and 3 weeks after fracture [(2.57 ±0.14) ng/ml,(3.47 ±0.26) ng/ml] were significantly higher than those of control group [(2.07 ±0.19)ng/ml,(2.77 ±0.29)ng/ml];the PINP at 1,2,and 3 weeks after fracture in the experimental group [(2 216.50 ±96.68)pg/ml,(2 692.33 ± 136.76)pg/ml,and (3 196.75 ± 221.90)pg/ml] were significantly higher than those of the control group [(1 969.50 ± 89.34) pg/ml,(2 241.33 ± 107.86) pg/ml,and (2 603.25 ± 361.60) pg/ml];the levels of TRACP5b [(11.58 ± 0.47) ng/ml] and CTX [(8.02 ± 0.40) ng/ml] at 1 week after fracture were higher than those of the control group [(10.33 ± 0.53) ng/ml,(7.11 ± 0.36) ng/ml] (P ＜ 0.05).Conclusion Gefitinib can promote the maturation of osteoblasts and the early reabsorption function of osteoclasts to induce bone formation in advance and accelerate the fracture healing.