In recent years, the bacteriophage Φ29 (Phi29) DNA polymerase has garnered increasing attention due to its high-fidelity amplification capacity at constant temperatures. To advance the industrial application of this type of isothermal polymerases, this study mined and characterized new enzymes from the microbial metagenome based on the known Phi29 DNA polymerase sequence. The results revealed that a new enzyme, Php29 DNA polymerase, was identified in the microbial metagenome with plants as the hosts. This enzyme exhibited higher strand displacement activity, with a 59.5% similarity to bacteriophage Φ29. Experimental validation demonstrated that the enzyme had 3'→5' exonuclease activity, and its amplification products can serve as substrates for further catalytic reactions. The discovery and validation of Php29 DNA polymerase gives insights into the future industrial application of isothermal polymerases.
DNA-Directed DNA Polymerase/metabolism* ; Bacillus Phages/genetics* ; Metagenome

Objective:To discuss the clinical curative effect of the treatment of bundling bone fragment with figure-of-eight suture through double bone tunnels in the chronic bong mallet finger.Methods:A retrospective analysis was performed on patients with chronic bong mallet fingers who underwent surgery in the Department of Hand and Foot Microsurgery of Nanjing Jiangbei Hospital from May 2021 to February 2023. During the procedure, made into transverse double bone tunnels on the base of the distal phalanx with 0. 8 mm Kirschner wire, bundling bone fragment with figure-of-eight suture through double bone tunnels, then fixed with Kirschner wire elastic compression. Removal of the Kirschner wire 3 weeks after the procedure, the active flexion and extension range of the joints of the affected finger and the corresponding finger were measured at the last follow-up, then the range of motion (ROM) of the distal interphalangeal joint (DIPJ) and total action movement (TAM) of the fingers (affected and healthy finger) were recorded. The curative effects were evaluated according to the TAM system of the American Association of Hand Surgeons, including 4 grades: excellent, good, fair, and poor. SPSS 13. 0 software was used for statistical analysis of the data, and the measurement data conforming to normal distribution were expressed as Mean ± SD. The ROM of DIPJ and TAM of the affected finger were compared with the corresponding healthy finger 6 months after surgery by a paired sample t test. P<0. 05 indicates that the difference is statistically significant. Results:A total of 30 patients (30 digits) were enrolled, including 19 males and 11 females, and the age ranged from 18 to 62 years old, with an average age of 31 years old. The time from injury to operation was 24 to 65 days (mean of 35 days) . According to Wehbe and Schneider classification, there were 8 cases of type Ⅰa, 6 cases of type Ⅰb, 7 cases of type Ⅱa, 5 cases of type Ⅱb, 2 cases of type Ⅲa and 2 cases of type Ⅲb. All incisions healed well, the intraoperative blood loss was minimal, with no infection. All 30 cases were followed up for 6 to 7 months. All fractures were well aligned and healed, malformed fingers were completely corrected, and no needle tunnel infection or needle breakage were observed. At the last follow-up, the differences of the ROM of the DIPJ [ (30. 2±3. 5) °vs. (30. 4±3. 3) °, t=2. 57, P=0. 463] and the TAM [ (235. 3± 3. 6) ° vs. (237. 7± 4. 2) °, t=1. 78, P= 0. 247 ] between the affected and healthy fingers were not statistically significant. Finger function assessment: 27 cases were excellent, and 3 cases were good, and the excellent and good rate was 100%. Conclusion:Satisfactory therapeutic outcome for the treatment of chronic bong mallet finger deformity can be achieved by bundling bone fragment with figure-of-eight suture through double bone tunnels. It is an effective and practical method.

Objective:To discuss the clinical curative effect of the treatment of bundling bone fragment with figure-of-eight suture through double bone tunnels in the chronic bony mallet finger.Methods:A retrospective analysis was performed on patients with chronic bony mallet fingers who underwent surgery in the Department of Hand and Foot Microsurgery of Nanjing Jiangbei Hospital from May 2021 to February 2023. During the procedure, made into transverse double bone tunnels on the base of the distal phalanx with 0.8 mm Kirschner wire, bundling bone fragment with figure-of-eight suture through double bone tunnels, then fixed with Kirschner wire elastic compression. Removal of the Kirschner wire 3 weeks after the procedure, the active flexion and extension range of the joints of the affected finger and the corresponding finger were measured at the last follow-up, then the range of motion (ROM) of the distal interphalangeal joint (DIPJ) and total action movement (TAM) of the fingers (affected and healthy finger) were recorded. The curative effects were evaluated according to the TAM system of the American Association of Hand Surgeons, including 4 grades: excellent, good, fair, and poor. SPSS 13.0 software was used for statistical analysis of the data, and the measurement data conforming to normal distribution were expressed as Mean±SD. The ROM of DIPJ and TAM of the affected finger were compared with the corresponding healthy finger 6 months after surgery by a paired sample t-test. P<0.05 indicated that the difference was statistically significant. Results:A total of 30 patients (30 digits) were enrolled, including 19 males and 11 females, and the age ranged from 18 to 62 years old, with an average age of 31 years old. The time from injury to operation was 24 to 65 days (mean of 35 days). According to Wehbe and Schneider classification, there were 8 cases of type Ⅰa, 6 cases of type Ⅰb, 7 cases of type Ⅱa, 5 cases of type Ⅱb, 2 cases of type Ⅲa and 2 cases of type Ⅲb. All incisions healed well, the intraoperative blood loss was minimal, with no infection. All 30 cases were followed up for 6 to 7 months. All fractures were well aligned and healed, malformed fingers were completely corrected, and no needle tunnel infection or needle breakage were observed. At the last follow-up, the differences of the ROM of the DIPJ [(30.2±3.5)° vs. (30.4±3.3)°, t=2.57, P=0.463] and the TAM [(235.3±3.6)° vs. (237.7±4.2)°, t=1.78, P=0.247] between the affected and healthy fingers were not statistically significant. Finger function assessment: 27 cases were excellent, and 3 cases were good, and the excellent and good rate was 100%. Conclusion:Satisfactory therapeutic outcome for the treatment of chronic bony mallet finger deformity can be achieved by bundling bone fragment with figure-of-eight suture through double bone tunnels. It is an effective and practical method.

Objective:To discuss the clinical curative effect of the treatment of bundling bone fragment with figure-of-eight suture through double bone tunnels in the chronic bong mallet finger.Methods:A retrospective analysis was performed on patients with chronic bong mallet fingers who underwent surgery in the Department of Hand and Foot Microsurgery of Nanjing Jiangbei Hospital from May 2021 to February 2023. During the procedure, made into transverse double bone tunnels on the base of the distal phalanx with 0. 8 mm Kirschner wire, bundling bone fragment with figure-of-eight suture through double bone tunnels, then fixed with Kirschner wire elastic compression. Removal of the Kirschner wire 3 weeks after the procedure, the active flexion and extension range of the joints of the affected finger and the corresponding finger were measured at the last follow-up, then the range of motion (ROM) of the distal interphalangeal joint (DIPJ) and total action movement (TAM) of the fingers (affected and healthy finger) were recorded. The curative effects were evaluated according to the TAM system of the American Association of Hand Surgeons, including 4 grades: excellent, good, fair, and poor. SPSS 13. 0 software was used for statistical analysis of the data, and the measurement data conforming to normal distribution were expressed as Mean ± SD. The ROM of DIPJ and TAM of the affected finger were compared with the corresponding healthy finger 6 months after surgery by a paired sample t test. P<0. 05 indicates that the difference is statistically significant. Results:A total of 30 patients (30 digits) were enrolled, including 19 males and 11 females, and the age ranged from 18 to 62 years old, with an average age of 31 years old. The time from injury to operation was 24 to 65 days (mean of 35 days) . According to Wehbe and Schneider classification, there were 8 cases of type Ⅰa, 6 cases of type Ⅰb, 7 cases of type Ⅱa, 5 cases of type Ⅱb, 2 cases of type Ⅲa and 2 cases of type Ⅲb. All incisions healed well, the intraoperative blood loss was minimal, with no infection. All 30 cases were followed up for 6 to 7 months. All fractures were well aligned and healed, malformed fingers were completely corrected, and no needle tunnel infection or needle breakage were observed. At the last follow-up, the differences of the ROM of the DIPJ [ (30. 2±3. 5) °vs. (30. 4±3. 3) °, t=2. 57, P=0. 463] and the TAM [ (235. 3± 3. 6) ° vs. (237. 7± 4. 2) °, t=1. 78, P= 0. 247 ] between the affected and healthy fingers were not statistically significant. Finger function assessment: 27 cases were excellent, and 3 cases were good, and the excellent and good rate was 100%. Conclusion:Satisfactory therapeutic outcome for the treatment of chronic bong mallet finger deformity can be achieved by bundling bone fragment with figure-of-eight suture through double bone tunnels. It is an effective and practical method.

Objective:To discuss the clinical curative effect of the treatment of bundling bone fragment with figure-of-eight suture through double bone tunnels in the chronic bony mallet finger.Methods:A retrospective analysis was performed on patients with chronic bony mallet fingers who underwent surgery in the Department of Hand and Foot Microsurgery of Nanjing Jiangbei Hospital from May 2021 to February 2023. During the procedure, made into transverse double bone tunnels on the base of the distal phalanx with 0.8 mm Kirschner wire, bundling bone fragment with figure-of-eight suture through double bone tunnels, then fixed with Kirschner wire elastic compression. Removal of the Kirschner wire 3 weeks after the procedure, the active flexion and extension range of the joints of the affected finger and the corresponding finger were measured at the last follow-up, then the range of motion (ROM) of the distal interphalangeal joint (DIPJ) and total action movement (TAM) of the fingers (affected and healthy finger) were recorded. The curative effects were evaluated according to the TAM system of the American Association of Hand Surgeons, including 4 grades: excellent, good, fair, and poor. SPSS 13.0 software was used for statistical analysis of the data, and the measurement data conforming to normal distribution were expressed as Mean±SD. The ROM of DIPJ and TAM of the affected finger were compared with the corresponding healthy finger 6 months after surgery by a paired sample t-test. P<0.05 indicated that the difference was statistically significant. Results:A total of 30 patients (30 digits) were enrolled, including 19 males and 11 females, and the age ranged from 18 to 62 years old, with an average age of 31 years old. The time from injury to operation was 24 to 65 days (mean of 35 days). According to Wehbe and Schneider classification, there were 8 cases of type Ⅰa, 6 cases of type Ⅰb, 7 cases of type Ⅱa, 5 cases of type Ⅱb, 2 cases of type Ⅲa and 2 cases of type Ⅲb. All incisions healed well, the intraoperative blood loss was minimal, with no infection. All 30 cases were followed up for 6 to 7 months. All fractures were well aligned and healed, malformed fingers were completely corrected, and no needle tunnel infection or needle breakage were observed. At the last follow-up, the differences of the ROM of the DIPJ [(30.2±3.5)° vs. (30.4±3.3)°, t=2.57, P=0.463] and the TAM [(235.3±3.6)° vs. (237.7±4.2)°, t=1.78, P=0.247] between the affected and healthy fingers were not statistically significant. Finger function assessment: 27 cases were excellent, and 3 cases were good, and the excellent and good rate was 100%. Conclusion:Satisfactory therapeutic outcome for the treatment of chronic bony mallet finger deformity can be achieved by bundling bone fragment with figure-of-eight suture through double bone tunnels. It is an effective and practical method.

Hypoxia-inducible factor (HIF-1α), a core transcription factor responding to changes in cellular oxygen levels, is closely associated with a wide range of physiological and pathological conditions. However, its differential impacts on vascular cell types and molecular programs modulating human vascular homeostasis and regeneration remain largely elusive. Here, we applied CRISPR/Cas9-mediated gene editing of human embryonic stem cells and directed differentiation to generate HIF-1α-deficient human vascular cells including vascular endothelial cells, vascular smooth muscle cells, and mesenchymal stem cells (MSCs), as a platform for discovering cell type-specific hypoxia-induced response mechanisms. Through comparative molecular profiling across cell types under normoxic and hypoxic conditions, we provide insight into the indispensable role of HIF-1α in the promotion of ischemic vascular regeneration. We found human MSCs to be the vascular cell type most susceptible to HIF-1α deficiency, and that transcriptional inactivation of ANKZF1, an effector of HIF-1α, impaired pro-angiogenic processes. Altogether, our findings deepen the understanding of HIF-1α in human angiogenesis and support further explorations of novel therapeutic strategies of vascular regeneration against ischemic damage.
Humans ; Vascular Endothelial Growth Factor A/metabolism* ; Endothelial Cells/metabolism* ; Transcription Factors/metabolism* ; Gene Expression Regulation ; Hypoxia/metabolism* ; Cell Hypoxia/physiology*

ObjectiveTo explore the impact of autonomic nerve function on motor function in patients with post-stroke depression (PSD) from the perspective of regional homogeneity (ReHo). MethodsFrom January to December, 2020, a total of 60 inpatients and outpatients with cerebral infarction in the Affiliated Brain Hospital of Nanjing Medical University were divided into control group (n = 30) and PSD group (n = 30). Two groups were assessed using Fugl-Meyer Assessment (FMA), modified Barthel Index (MBI) and Hamilton Depression Scale (HAMD). Heart rate variability (HRV) was measured. Ten patients in each group were selected randomly to undergo resting state functional magnetic resonance imaging (rs-fMRI) to calculate ReHo. ResultsAll HRV indices were lower in PSD group than in the control group (|t| > 2.092, P < 0.05). In PSD group, FMA and MBI scores showed positive correlations with 24-hour standard deviation of normal-to-normal R-R intervals (SDNN), the root mean square of successive differences between normal heartbeats over 24 hours (RMSSD), the percentage of differences between adjacent normal R-R intervals over 24 hours that were greater than 50 ms (PNN50), total power (TP), very low frequency power (VLF) and low frequency power (LF) (r > 0.394, P < 0.05), and showed negative correlations with HAMD scores (|r| > 0.919, P < 0.001). HAMD scores in PSD group were negatively correlated with SDNN, RMSSD, PNN50, TP and VLF (|r| > 0.769, P < 0.001). Compared with the control group, the ReHo increased in PSD group in the right rectus gyrus (142 voxels, t = 6.575), the left medial and paracingulate gyri (204 voxels, t = 4.925) (GRF correction, P_-Voxel < 0.005，P_-Cluster < 0.05); and reduced in the right cerebellum (191 voxels, t = -6.487), the left middle temporal gyrus (140 voxels, t = -5.516), and the left precentral gyrus (119 voxels, t = -4.764) (GRF correction, P_-Voxel < 0.005，P_-Cluster < 0.05) in PSD group. ConclusionAutonomic nerve function is related to motor dysfunction in patients with PSD. The modulation of emotional, cognitive and motor brain regions by the autonomic nervous system may play a role in influencing the motor function in patients with PSD.

ObjectiveTo investigate whether paeonol exerts a protective effect on mice with alcoholic liver injury by regulating the takeda G-protein-coupled receptor 5 （TGR5）/protein kinase A （PKA）/cAMP response binding element （CREB） signaling pathway mediated by Eubacterium. MethodC57BL/6 mice were randomly divided into five groups： normal group， model group， paeonol group （480 mg·kg^-1）， antibiotic group （Abs group）， and antibiotic + paeonol group. Lieber-DeCarli liquid was used to feed C57BL/6 mice on the second day of modeling for 10 days. The blood lipids， liver function， inflammatory factors， and oxidative stress levels in mice were measured. Hematoxylin-eosin staining （HE） and oil red O staining were used to observe the morphological changes and fat accumulation in liver tissue. 16S rDNA sequencing was used to detect the diversity of intestinal microbiota in the blank， model， and paeanol groups. Western blot was used to detect the effect of paeonol on the expression levels of protein related to the signaling pathway of atresia band protein 1 （ZO-1）， Claudin-1， and TGR5/PKA/CREB in mouse ileal tissue. ResultCompared with those in the blank group， the blood lipids， liver function， oxidative stress levels， and the expression of inflammatory factors in the model group increased （P<0.01）， and the liver fat vacuoles were obvious. The ileal mucosa was seriously damaged， and the protein contents of ZO-1， Claudin-1， and TGR5/PKA/CREB in the ileal tissue decreased significantly （P<0.01）. The intestinal microbiota changed， and the proteobacteria phylum increased significantly. The ratio of Bacteroidetes to Firmicutes decreased. The relative abundance of Dubosiella newyorkensis， Lactobacillus， Bifidobacterium， and other genera decreased， while the relative abundance of Escherichia-Shigella， Morganella， Providencia， and Proteus increased significantly. Compared with the model group， paeonol significantly reduced the blood lipids， liver function， oxidative stress levels， and expression of inflammatory factors in mice with alcohol diet-induced liver injury （P<0.05）， decreased liver fat vacuoles， improved and restored the ileal intestinal barrier， and restored the normal structure of hepatocytes and ileal cells. The intestinal microbiota disorder caused by alcohol was improved， and the relative abundance of beneficial bacteria such as Eubacterium spp. was increased. The protein expression levels of ZO-1， Claudin-1， and TGR5/PKA/CREB in ileal tissue were increased （P<0.05）. ConclusionPaeonol has a protective effect on alcoholic liver injury in mice， and the mechanism of action is achieved by regulating the Eubacterium-mediated TGR5/PKA/CREB signaling pathway to ensure anti-inflammatory effect and improve the intestinal barrier.

Objective To observe the effect of ultrasound-guided anterior quadratus lumborum block at lateral supra-arcuate ligament(QLB-LSAL)and transversus abdominis plane block(TAPB)on analgesia and recovery quality after laparoscopic partial hepatectomy(LPH).Methods Fifty-eight patients underwent elective LPH were selected and divided into the quadratus lumborum group or the transversus abdominis group randomly,with 29 patients in each group.The quadratus lumborum group received bilateral QLB-LSAL,and the transversus abdominis group received bilateral subcostal TAPB block before surgery.Both groups received 20 mL of 0.33%ropivacaine on each side.All patients used patient-controlled intravenous analgesia(PCIA)postoperatively.The numeric rating scale(NRS)scores for rest and movement were recorded at 2,4,6,12,24 and 48 hours postoperatively,as well as the Quality of Recovery-15(QoR-15)scores at 1 day preoperatively,1 and 3 days postoperatively.The perioperative anesthetic agent consumption,PCIA pressing frequency,remedial analgesia use in 48 h,postoperative nausea and vomiting(PONV)incidence and time of first out-of-bed mobilization were also recorded.Results Compared with the transversus abdominis group,the quadratus lumborum group had lower movement NRS scores at 2,4,6,12,24 and 48 hours postoperatively,and lower rest NRS scores at 2,4,6,12 and 24 hours postoperatively(P＜0.05).The quadratus lumborum group had higher QoR-15 scores at 1 and 3 days postoperatively(P＜0.05).Patients in the quadratus lumborum group had reduced perioperative remifentanil and sufentanil consumption,postoperative 48-hour rescue analgesia use,PCIA pressing frequency,PONV incidence and time of first out-of-bed mobilization(P＜0.05).Conclusion QLB-LSAL block provides superior analgesic effects and recovery quality compared to TAPB block after LPH.

With the continuous development of medical science and the widespread use of antibiotics,the problem of bacterial resistance is increasing,especially the increasing carbapenem-resistant Enterobacteriaceae(CRE)infection,and the high mortality rate,which brings great challenges to clinical treatment.In this paper,the mechanism of drug resistance,existing antibac-terial drugs,and exploratory treatment options for CRE are reviewed,and the research progress in treating CRE infection is dis-cussed to provide more reliable evidence and a theoretical basis for clinical practice.