[Clinical effects of free descending branch tissue flap of lateral circumflex femoral artery in repairing Wagner grade 3 or 4 diabetic foot wounds].

Objective: To investigate the clinical effects of free descending branch tissue flap of lateral circumflex femoral artery in repairing Wagner grade 3 or 4 diabetic foot wounds. Methods: A retrospective observational study was conducted. From October 2019 to January 2022, 12 patients (10 males and 2 females, aged 38-66 years, with an average of 52 years) with diabetic foot wounds who met the inclusion criteria were admitted to the First People's Hospital of Yulin, of whom 5 cases being Wagner grade 3 and 7 cases being Wagner grade 4. Wound debridement and vacuum sealing drainage were performed to control infection, and the wound area after debridement was between 13 cm×6 cm and 28 cm×11 cm. The wounds were repaired with free descending branch tissue flaps of lateral circumflex femoral artery. Among them, the wounds of 4 cases were repaired with single flap of musculocutaneous branch or intermuscular branch of descending branch tissue flap of lateral circumflex femoral artery, 1 case with lobulated flap, 1 case with bilateral tandem flap, and 6 cases with chimeric lateral femoral muscle flap. The area of resected tissue flap was 9 cm×7 cm to 21 cm×10 cm; end-to-end anastomosis was selected between vessels in the donor sites and the recipient sites, and pressurized treatment should be performed if necessary. The donor site wounds of 10 patients were treated with cosmetic tension-relieving suture, and the donor site wounds of 2 patients were repaired with split-thickness skin graft from head. After operation, the condition of wound repair was recorded. After wound healing, the level of 2-hour postprandial blood glucose of patients was measured. During follow-up, the wound healing of the recipient site and scar formation of the donor site were observed. Before and 6 months after operation, computer tomography angiography was used to detect and compare the blood perfusion of the affected limb. At the last follow-up, the walking ability of the patients was recorded. Results: The wounds of 9 patients healed well after operation; the flaps in the recipient site of 2 patients were infected on postoperative day 5 after surgery and were repaired by suturing in stage Ⅱ after open drainage and inflammation control; the distal end of the grafted tandem flap in 1 patient gradually developed purple necrosis on postoperative day 4 and was repaired with a skin graft after debridement. After wound healing, the 2-hour postprandial blood glucose value of all patients was controlled in the range of 8-12 mmol/L. During Follow-up of 6 months to 1 year, 3 patients had partial ulceration in the plantar compression area, which healed after decompression combined with dressing change or flap repair, while the other 9 patients had no ulceration in the recipient area; the appearance of the affected foot was plump after wound healing, the transplanted flaps had good blood supply and good fit with the surrounding tissue, and were wear-resistant, but insensitive. During follow-up, only linear scar remained in the donor area of direct suture in 10 cases, and the skin grafts in the donor area were completely survived in 2 cases. Compared with the patency of the main blood vessels of the ankle joint segment of the affected limb before operation, the vascular network of the flap transplantation area in the affected foot was formed 6 months after operation. At the last follow-up, 8 patients could walk independently, and 4 patients could walk with crutches. Conclusions: The free descending branch tissue flap of lateral circumflex femoral artery is effective in repairing Wagner grade 3 or 4 diabetic foot wounds, which can shorten the course of disease and improve local blood flow.

[Aluminuminduced impairment in primary cultured rat choroid plexus epithelial cells].

OBJECTIVE: To investigate the impairment in primary cultured rat choroid plexus epithelial cells (CPECs)induced by aluminum. METHODS: The choroid plexus isolated from Sprague-Dawley rats 14 days old was cut into pieces and digested by trypsin in the sterile area. The obtained single cells were cultured in DMEM with 1% epidermal growth factor and 20% fetal calf serum. Five days later, immunohistochemistry with anti-transthyretin antibody was used to identify the purity of cultured cells. The well-grown cells were treated with aluminum lactate at different concentrations (0, 100, 400, and 1 600 µmol/L for control, lowdose, mediumdose, and highdose groups). Fortyeight hours later, the cell viability, apoptotic rate, level of reactive oxygen species (ROS), and activity of superoxide dismutase (SOD)were measured in each group to evaluate the impairment in primary cultured rat CPECs by aluminum. RESULTS: More than 95% of the cultured cells were identified as CPECs. The medium-and high-dose groups had significantly lower cell viability than the control group(86.74%±4.03% vs 100%, P<0.01; 81.90%±9.17% vs 100%, P<0.01). The high-dose group had significantly lower cell viability than the lowdose group (81.90%±9.17% vs 92.92%±8.81%, P<0.01). The medium-and high-dose groups had significantly higher apoptotic rates than the control group (7.26%±0.99% vs 1.29%±0.03%, P<0.01; 22.25%±1.55% vs 1.29%±0.03%, P<0.01)and the low-dose group (7.26%±0.99% vs 1.68%±0.27%, P<0.01; 22.25%±1.55% vs 1.68%±0.27%, P<0.01). The high-dose group had a significantly higher apoptotic rate than the medium-dose group (22.25%±1.55% vs 7.26%±0.99%, P<0.01). The mediumand high-dose groups had significantly higher fluorescence intensity of ROS than the control group (22.23%±0.41% vs 17.24%±0.09%, P<0.05; 25.10%±1.13% vs 17.24%±0.09%, P<0.05)and the lowdose group (22.23%±0.41% vs 18.31%±0.21%, P<0.05; 25.10%±1.13% vs 18.31%±0.21%, P<0.05). The highdose group had significantly higher fluorescence intensity of ROS than the mediumdose group (25.10%±1.13% vs 22.23%±0.41%, P< 0.05). The low-, medium-and high-dose groups had significantly lower SOD activity than the control group[(28.65±0.74)U/g Hb vs (37.35±1.05)U/g Hb, P<0.05; (22.75±1.94)U/g Hb vs (37.35±1.05)U/g Hb, P<0.05; (13.29±0.64)U/g Hb vs(37.35±1.05)U/g Hb, P<0.05]. The medium-and high-dose groups had significantly lower SOD activity than the low-dose group[(22.75±1.94)U/g Hb vs(28.65±0.74)U/g Hb, P<0.05; (13.29±0.64)U/g Hb vs (28.65±0.74)U/g Hb, P<0.05], while the high-dose group had had significantly lower SOD activity than the medium-dose group[(13.29±0.64)U/g Hb vs (22.75±1.94)U/g Hb, P<0.05]. There were no significant differences in cell viability, apoptotic rate, level of ROS, or activity of SOD between any other two groups (P>0.05). CONCLUSION: Aluminum lactate may induce impairment in primary cultured rat CPECs. It reduces the cell viability, elevates the apoptotic rate, and causes oxidative stress.

[Effects of subchronic aluminum lactate exposure on learning and memory and transportation of Aß in blood-cerebrospinal fluid in rats].

OBJECTIVE: To investigate the effects of aluminum lactate exposure on learning and memory and the transportation of amyloid-beta peptides(Aß) in cerebrospinal fluid in rats. METHODS: A total of 80 male Sprague-Dawley rats were randomly divided into solvent control(distilled water) group and low-, medium-, and high-dose aluminum poisoning groups(10, 30, and 90 mg/kg aluminum lactate), with 20 rats in each group, and the poisoning procedure was performed by gavage for 2 months. The Morris water maze test was used to test the rats' learning and memory, Western blot was used to measure the expression level of low-density lipoprotein receptor protein-1(LRP-1) in rats' choroid plexus, and enzyme-linked immunosorbent assay(ELISA) was used to measure the content of Aß in the cerebrospinal fluid and plasma. RESULTS: The Morris water maze test showed that in the place navigation test, with the increasing training time, the escape latency was significantly shortened in each group and showed significant differences between any two groups(P<0.05). In the spatial probe test, the time spent in target quadrant in the medium-and high-dose groups was 11.52±1.56 s and 10.43±5.27 s, respectively, which was significantly shorter than that in the control group and the low-dose group(15.81±3.01 s and 13.91±2.17 s)(P<0.05). The numbers of platform crossings in the medium-and high-dose groups were 2.64±1.39 and 1.50±0.76, respectively, which were significantly lower than those in the control group and the low-dose group(4.29±0.914 and 3.56±1.38)(P<0.05). The results of ELISA showed that the medium-and high-dose groups had significant increases in the content of Aß1-42 in cerebrospinal fluid(320.35±84.82 pg/ml and 327.68±67.51 pg/ml), which was significantly higher than that in the control group(203.46±74.36 pg/ml) (P<0.05). The content of Aß1-42 in plasma showed no significant difference between any two groups(P>0.05), and that of Aß1-40 in cerebrospinal fluid and plasma also showed no significant difference between any two groups(P>0.05). The results of Western blot showed that the high-dose group had significantly lower protein expression of LRP-1 than the control group and the low-and medium-dose groups(0.57±0.21 vs 1.00±0.00/0.79±0.15/0.95±0.24, P<0.05). CONCLUSION: Subchronic aluminum exposure may reduce learning and memory in rats, and the accumulation of Aß in cerebrospinal fluid may be related to the reduced protein expression of LRP-1 in the choroid plexus, suggesting that aluminum affects learning and memory in rats through reducing the protein expression of LRP-1, influencing the transportation of Aß, and leading to the accumulation of Aß.

Cloning and sequence analysis of the Blumea balsamifera DC farnesyl diphosphate synthase gene.

Blumea balsamifera DC is a member of the Compositae family and is frequently used as traditional Chinese medicine. Blumea balsamifera is rich in monoterpenes, which possess a variety of pharmacological activities, such as antioxidant, anti-bacteria, and anti-viral activities. Farnesyl diphosphate synthase (FPS) is a key enzyme in the biosynthetic pathway of terpenes, playing an important regulatory role in plant growth, such as resistance and secondary metabolism. Based on the conserved oligo amino acid residues of published FPS genes from other higher plant species, a cDNA sequence, designated BbFPS, was isolated from B. balsamifera DC using polymerase chain reaction. The clones were an average of 1.6 kb and contained an open reading frame that predicted a polypeptide of 342 amino acids with 89.07% identity to FPS from other plants. The deduced amino acid sequence was dominated by hydrophobic regions and contained 2 highly conserved DDxxD motifs that are essential for proper functioning of FPS. Phylogenetic analysis indicated that FPS grouped with other composite families. Prediction of secondary structure and subcellular localization suggested that alpha helices made up 70% of the amino acids of the sequence.

Genetic diversity of the Chinese traditional herb Blumea balsamifera (Asteraceae) based on AFLP markers.

Blumea balsamifera is a commercially important medicinal herb in China and other parts of Asia. It is used to produce borneol. This plant grows in the wild, but resources have diminished greatly in recent years. We examined the genetic diversity of this species to help develop conservation strategies; 35 plants from five provinces were analyzed using AFLPs. Eight AFLP primer combinations generated 1367 fragments, giving a mean of 172 fragments per primer combination. Polymorphism in the germplasm analysis was found for 1360 (99.48%) of the fragments, of which 264 (19.27%) fragments were unique (accession specific) and 423 (25.33%) of the fragments were rare (present in less than 10% of the accessions). The polymorphic fragments were used to group the accessions in a UPGMA phenogram. Most grouping was geographical. In general, accessions coming from Guizhou and Guangxi showed higher diversities as these accessions were scattered in different groups. The genetic distance estimated by Jaccard similarity coefficient index showed low variability among genotypes (coefficient value ranged from 0.60 to 0.95). More attention should be given to the study and conservation of the biodiversity of this economically important genus.