BACKGROUND:Nowadays, gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. Its key is to choose proper cel s, genes and vectors. OBJECTIVE:To use recombinant adeno-associated virus mediatedβ-nerve growth factor (β-NGF) to transfect rat bone marrow-derived endothelial progenitor cel s in vitro, and to investigate the effect ofβ-NGF expression on the proliferation of endothelial progenitor cel s. METHODS:The endothelial progenitor cel s were isolated, cultured and identified from the bone marrow of rats. Empty vector or recombinant adenovirus-associated virus containingβ-NGF gene was transferred into endothelial progenitor cel s. We examined the transfection efficiency by fluorescence expression of green fluorescent protein. Expression ofβ-NGF protein was detected using ELISA, and its effect on the proliferation of endothelial progenitor cel s was determined using MTT method. RESULTS AND CONCLUSION:Rat endothelial progenitor cel s were isolated and cultured successful y in vitro and were identified positive by the function of cel s and immunofluorescence staining. The endothelial progenitor cel s were infected directly by the recombinant adenovirus-associated virus containingβ-NGF gene with an efficiency of 65.3%.β-NGF protein was detected in the culture supernatant of transfected endothelial progenitor cel s, which reached a high level at 10 days after gene transfection. Furthermore, there was noβ-NGF protein in the blank and empty vector groups. After transfection, the proliferative ability of endothelial progenitor cel s was increased, which was significantly higher than the blank and empty vector groups (P<0.05). But there was no difference between the latter two groups (P>0.05). These findings suggest that recombinant adenovirus-associated virus containingβ-NGF gene can be successful y transferred into rat bone marrow-derived endothelial progenitor cel s and promote the proliferation of endothelial progenitor cel s.

24 new cDNA sequences encoding cytochrome P450 were amplified respectively from deltamethrin susceptible and -resistant strains of the mosquito, Culex pipiens pallens, with a pair of degenerate primers according to the conservative amino acid sequences of CYP4 in insects by RT-PCR. Studies of molecular systematics show that the 24 new genes (alleles) belong to CYP4C, CYP4D, CYP4H and CYP4J subfamilies of the CYP4 family, and they were named by Cytochrome P450 Nomenclature Committee. Among the new genes (alleles), CYP4C23 may be a pseudogene, CYP4H13 has a retained intron 58 nucleotides in length, and CYP4J4V1 has a stop coden (TAG) in frame near the 3'-end.

Objective To investigate the clinical efficacy of deep inferior epigastric perforator (DIEP) flap in the repair of soft tissue defects of the calf.Methods From January,2015 to January,2017,the DIEP flap was used to repair the soft tissue defect of calf in 11 patients,which were 7 males and 4 females,aged 23-62 years,with an average age of 42 years.Among them,there were 5 cases of anterior tibial soft tissue defect and 6 cases of posterior tibial soft tissue defect.The size of soft tissue defects ranged from 9.0 cm×5.0 cm to 32.0 cm×12.0 cm.The wounds were heavily polluted and debridement exploration combined VSD surgery in emergency.After 7 to 14 days,free deep inferior epigastric perforator flap was used to repaire.The flap was cut in the range of 10.0 cm×6.0 cm-34.0 cm×13.0 cm.Nine cases of abdominal donor sutured direct,and a small part of 2 cases to take thigh thick skin graft repair.Results All the flaps survived.In only 1 patient,the distal part of the flap was necrotic and healed after appropriate treatment.Eleven patients were followed-up for 3-18 months (average of 10 months).The shape of the flap was similar to that of the affected area.The texture was soft and not bloated.Conclusion The DIEP flap can provide reliable blood supply,large cutting area and flexible design.It can be used to repair soft tissue defect of calf and obtain satisfactory clinical results.

Objective:To evaluate the effect of the free anterolateral thigh flap with nerve and the sural neurocutaneous flap on ankle and foot tissue defect, and to provide guidance for clinical selection.Methods:From May 2016 to May 2019, we reviewed and analyzed the data of patients with soft tissue injury of ankle and foot repaired by flap from Microsurgery Repair and Reconstruction Center, Traditional Chinese Medicine Hospital Affiliated to Southwest Medical University. According to the surgical method, they were divided into the free anterolateral thigh flap with nerve group and the sural neurocutaneous flap group. The survival rate of the 2 groups of skin flaps were calculated. According to the trial standard for evaluation of upper limb function in Chinese Medical Association Hand Surgery Society, the flaps were rated as excellent, good, and acceptable. And the excellent and good rates were counted. The incidence of regional complications of the 2 groups were counted. SPSS 26.0 software was used for analysis, the measurement data were subjected to t-test, expressed as mean±standard deviation; the categorical variables were subjected to chi-square test or Fisher’s exact probability method, and P<0.05 indicated that the difference was statistically significant. Results:A total of 59 patients with soft tissue defects in the foot and ankle were included. Among them, 21 cases were repaired with free anterolateral thigh flaps with nerve (group A), 12 males and 9 females, with an average age of 42.1 years, and the defect area was 4 cm×6 cm-11 cm ×16 cm, followed up for an average of 12.3 months after surgery; 17 cases of sural neurocutaneous flap repair (group B), 11 males and 6 females, average age 45.3 years, defect area 5 cm×5 cm-10 cm× 14 cm, the average follow-up was 11.3 months. There were no statistically significant differences in age, gender and cause of injury between the two groups ( P<0.05). The area of the flaps in group A and group B were (53.0±12.2) cm 2 and (46.4±9.62) cm 2, respectively. There was no significant difference between the two groups ( t=1.824, P=0.076). The flaps were all survived in both groups. Among them, 2 cases of group A had partial necrosis at the distal end of the flaps and healed after post-treatment; 1 case of group B had vascular crisis after operation, and the flaps survived after exploratory decompression. The first-stage survival rate of skin flaps in group A was 90.5% (19/21), and the first-stage survival rate of skin flaps in group B was 94.1% (16/17). The difference was not statistically significant ( P=1.000). According to the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Society, the skin flaps in group A were excellent in 10 cases, good in 7 cases, and fair in 4 cases, with an excellent and good rate of 81.0% (17/21). In group B, skin flaps were excellent in 3 cases and good in 3 cases. Of the 11 cases, the excellent and good rate was 35.3% (6/17), and the difference was statistically significant ( P=0.007). The flap donor area in group A healed well without related complications. In group B, 2 patients (11.8%) developed scar contracture deformity after the flap donor area, which affected calf function, and the function recovered after the second-stage scar was released. The incidence of complications in the region was not statistically significant ( P=0.193). Conclusions:Both the free anterolateral thigh flap with nerve and the sural neurocutaneous flap can repair the soft tissue defect of foot and ankle with high survival rate. But the sural neurocutaneous flap has a higher good rate than the free anterolateral thigh flap with nerve, and the donor site is affected smaller.

Objective:To evaluate the effect of the free anterolateral thigh flap with nerve and the sural neurocutaneous flap on ankle and foot tissue defect, and to provide guidance for clinical selection.Methods:From May 2016 to May 2019, we reviewed and analyzed the data of patients with soft tissue injury of ankle and foot repaired by flap from Microsurgery Repair and Reconstruction Center, Traditional Chinese Medicine Hospital Affiliated to Southwest Medical University. According to the surgical method, they were divided into the free anterolateral thigh flap with nerve group and the sural neurocutaneous flap group. The survival rate of the 2 groups of skin flaps were calculated. According to the trial standard for evaluation of upper limb function in Chinese Medical Association Hand Surgery Society, the flaps were rated as excellent, good, and acceptable. And the excellent and good rates were counted. The incidence of regional complications of the 2 groups were counted. SPSS 26.0 software was used for analysis, the measurement data were subjected to t-test, expressed as mean±standard deviation; the categorical variables were subjected to chi-square test or Fisher’s exact probability method, and P<0.05 indicated that the difference was statistically significant. Results:A total of 59 patients with soft tissue defects in the foot and ankle were included. Among them, 21 cases were repaired with free anterolateral thigh flaps with nerve (group A), 12 males and 9 females, with an average age of 42.1 years, and the defect area was 4 cm×6 cm-11 cm ×16 cm, followed up for an average of 12.3 months after surgery; 17 cases of sural neurocutaneous flap repair (group B), 11 males and 6 females, average age 45.3 years, defect area 5 cm×5 cm-10 cm× 14 cm, the average follow-up was 11.3 months. There were no statistically significant differences in age, gender and cause of injury between the two groups ( P<0.05). The area of the flaps in group A and group B were (53.0±12.2) cm 2 and (46.4±9.62) cm 2, respectively. There was no significant difference between the two groups ( t=1.824, P=0.076). The flaps were all survived in both groups. Among them, 2 cases of group A had partial necrosis at the distal end of the flaps and healed after post-treatment; 1 case of group B had vascular crisis after operation, and the flaps survived after exploratory decompression. The first-stage survival rate of skin flaps in group A was 90.5% (19/21), and the first-stage survival rate of skin flaps in group B was 94.1% (16/17). The difference was not statistically significant ( P=1.000). According to the trial standard of upper limb function evaluation of the Chinese Medical Association Hand Surgery Society, the skin flaps in group A were excellent in 10 cases, good in 7 cases, and fair in 4 cases, with an excellent and good rate of 81.0% (17/21). In group B, skin flaps were excellent in 3 cases and good in 3 cases. Of the 11 cases, the excellent and good rate was 35.3% (6/17), and the difference was statistically significant ( P=0.007). The flap donor area in group A healed well without related complications. In group B, 2 patients (11.8%) developed scar contracture deformity after the flap donor area, which affected calf function, and the function recovered after the second-stage scar was released. The incidence of complications in the region was not statistically significant ( P=0.193). Conclusions:Both the free anterolateral thigh flap with nerve and the sural neurocutaneous flap can repair the soft tissue defect of foot and ankle with high survival rate. But the sural neurocutaneous flap has a higher good rate than the free anterolateral thigh flap with nerve, and the donor site is affected smaller.

Molecular knowledge of human gastric corpus epithelium remains incomplete. Here, by integrated analyses using single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and single-cell assay for transposase accessible chromatin sequencing (scATAC-seq) techniques, we uncovered the spatially resolved expression landscape and gene-regulatory network of human gastric corpus epithelium. Specifically, we identified a stem/progenitor cell population in the isthmus of human gastric corpus, where EGF and WNT signaling pathways were activated. Meanwhile, LGR4, but not LGR5, was responsible for the activation of WNT signaling pathway. Importantly, FABP5 and NME1 were identified and validated as crucial for both normal gastric stem/progenitor cells and gastric cancer cells. Finally, we explored the epigenetic regulation of critical genes for gastric corpus epithelium at chromatin state level, and identified several important cell-type-specific transcription factors. In summary, our work provides novel insights to systematically understand the cellular diversity and homeostasis of human gastric corpus epithelium in vivo.
Humans ; Epigenesis, Genetic ; Gastric Mucosa/metabolism* ; Chromatin/metabolism* ; Stem Cells ; Epithelium/metabolism* ; Fatty Acid-Binding Proteins/metabolism*