ObjectiveTo determine the optimal combination of the effective monomer components "quercetin-kaempferol-abietic acid-boswellic acid" in Fujin Shengjisan for promoting diabetic ulcer （DU） wound healing through uniform design， thereby achieving the modern application of the ancient formula. MethodsFollowing the principle of "uniform design-pharmacodynamic experiment-mathematical modeling and model verification"， the U₁₄（14⁵） uniform design table was adopted.The four monomer components of Chinese medicine were considered as the independent variables， and the proliferation rate of human umbilical vein endothelial cells （HUVECs） induced by glucose was used as the pharmacodynamic indicator. A mathematical model was constructed using DPS software to correlate the effective monomer components with the pharmacodynamic indicator. The results of uniform design were verified through CCK-8 assay， cell scratch healing， tube formation， Western blot， and Real-time PCR. ResultsAmong the 14 compatibility groups， compared with the high-glucose model group， compound compatibility group 6 showed the strongest proliferation effect and statistical significance （P<0.05）. Four quadratic polynomial regression equations （Y₁-Y₄） were obtained through DPS modeling. Considering the model's fit， stability， and practical application， equations Y₁-Y₃ were selected for the follow-up verification. To ensure experiment reproducibility， group 6 was used for validation. Group 6 and equations Y₁-Y₃ were renamed as compound prescription ① to compound prescription④， respectively， to represent the modern application of the ancient FJSJ Powder through compatibility of monomer components. Verification experiments showed that in the CCK-8， scratch healing， and tube formation assays， the cell viability， wound healing rate， and tube formation number of HUVECs stimulated with 50 mmol·L^-1 glucose were significantly reduced compared with the blank group. Moreover， the expression levels of angiogenesis-related cytokines， vascular endothelial growth factor （VEGF） and fibroblast growth factor 2 （FGF2）， and CD31 secretion were significantly down-regulated. However， after intervention with compound prescriptions ① to ④， compound prescriptions ① and ③ significantly improved the biological functions of HUVECs induced by 50 mmol·L^-1 glucose. Further analysis of the regression coefficients of compound prescriptions ① and ③， and the relative dose ratios of each monomer component， indicated that abietic acid， quercetin， and boswellic acid promoted angiogenesis of HUVECs in the high glucose environment， with a major effect （positive partial correlation coefficients， all > 0.9）. Abietic acid and boswellic acid， as well as kaempferol and boswellic acid， promoted angiogenesis in HUVECs through interaction （positive partial correlation coefficients）. ConclusionCompound prescriptions ① and ③ are the optimal combinations. They can reverse the inhibitory effects of high glucose， stimulate the proliferation， migration， and tube formation abilities of HUVECs in a high glucose environment， and promote the expression of vascular endothelial growth factorA（VEGFA）， FGF2， and CD31， thereby promoting angiogenesis and facilitating DU wound healing. This finding not only confirms the good reproducibility and feasibility of compound prescriptions ① and ③ but also provides new insights and methods for the rational construction of mathematical models to further study the compatibility theory of Chinese medicine.

Objective To investigate the expression of targeted regulation of microRNA-146a (miR-146a) on E-box zinc finger protein 1 gene (ZEB1) and its related molecular mechanisms involved in neuroprotection and autophagy inhibition in a rat model of cerebral hemorrhage. Methods Forty 8-week-old male SD rats were randomly divided into four groups: sham operation group, model group, miR-146a overexpression group and miR-146a underexpression group, with 10 rats in each group. Except for the sham operation group, the other three groups were induced to establish an aneurysmal spontaneous intracerebral hemorrhage rat model using type VII collagenase. Rats of four groups were fed a high-salt diet for 6 weeks and euthanized after an additional feeding for 20 weeks. Neurological function [modified neurologic severity score (mNSS)] and brain water content were compared among the four groups. FJC staining was used to detect the number of degenerated neurons; TUNEL staining was used to detect apoptosis; hematoxylin-eosin (HE) staining was used to observe pathological changes in brain tissue; quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-146a and ZEB1 mRNA; Western blot was used to detect the expression of ZEB1, autophagy-related proteins human recombinant autophagy effector protein (Beclin 1) and microtubule-associated protein 1 light chain 3 (LC3). Results Compared to the sham surgery group, the model group showed a significant increase in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was significantly decreased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1and LC3 proteins, were significantly increased in the model group (P < 0.05). Compared to the model group, the miR-146a overexpression group exhibited a significant decrease in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was significantly increased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1 and LC3 proteins were significantly reduced in the miR-146a overexpression group (P < 0.05). Compared to the model group, the miR-146a low-expression group showed a further increase in mNSS score, brain water content, the number of degenerated neurons and cell apoptosis rate, and the expression level of miR-146a was further decreased, while the expressions of ZEB1 mRNA, as well as ZEB1, Beclin 1 and LC3 proteins were further elevated in the miR-146a low-expression group (P < 0.05). Conclusion Upregulating miR-146a can exert neuroprotective effects by targeting and inhibiting ZEB1 gene and protein expression, as well as regulating autophagy activity.

Objective:To explore the surgical technique and clinical effect of pressure boost in repairing soft tissue defects of limbs with thinned anterolateral thigh perforator flap (ALTP) .Methods:From January, 2015 to December, 2018, 18 cases with soft tissue defects of limbs with various damages of blood vessels and nerves with explosure of tendon and bone. There were 13 males and 5 females aged between 18 to 56 (averaged of 36.3) years, which were 6 defects in shank, 4 in foot and ankle, 5 in forearm, and 3 in hand. The soft tissue defect area was 7 cm ×12 cm to 13 cm ×30 cm. Thinned ALTP was used to repair the wound surface. The perforating vessels of the distal flap were anastomosed with one branch of the internal vessel pedicle flap to increase the pressure hence the blood supply of the distal region. The donor sites were sutured directly or covered by skin graft. Followed-up was conducted by 1-2 monthly clinic visits and telephone or on-line review to check the flap survival and recovery of functions.Results:All flaps survived without arterial or venous crisis. One flap had partial necrosis at the distal end, and healed after dressing change. One case had a swelling flap due to a congestion beneath the flap. The wound achieved primary healing after removal of sutures, ligation of subcutaneous vessels and drainage of hematoma. All patients were followed-up for 6 to 18 (average, 9.5) months. All flaps had good appearance and texture. After rehabilitation treatment, most of the joint activity had been recovered: extension and flexion of wrists joints ranged 60°-80°, 70°-80° for metacarpophalangeal joints and 40°-60° for ankle joints. One patient underwent ankle joint dorsiflexion function reconstruction and flap thinning at 6 months after operation due to the defects of most of the extensor tendon.Conclusion:During the use of free ALTP to repair soft tissue defect of limbs, application of the technique of pressure boost is able to increase blood supply to the distal region of flap. It helps to reduce the incidence of infection and necrosis at the edge of the flap.

Objective To perform the cloning of the gene encoding Schistosoma japonicum Chinese-strain hypoxanthine-guanine phosphoribosyltransferase(HGPRT)and its expression in Escherichia coli.MethodsA couple of primers were designed with the BamHI restriction endonuclease site introduced in forward primer and SalI in reverse primer.Total RNA was isolated from adult worms of S.japonicum Chinese-strain(Anhui-strain,Sjc-A)and the SjcHGPRT gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR).The PCR product and the prokaryotic expression vector pET28a were digested by both restriction endonucleases BamHI and SalI.The target DNA fragments were purified and cloned properly into pET28a.After identification by en-donucleases digestion,PCR and sequencing,the recombinant plasmid pET28a-SjcHG PRT was transformed into competent E.coli BL21 and expressed in the presence of IPTG.Results pET28a-SjHGPRT was sequenced and shown to be 99% and 83% identical in deduced amino acid sequence to that of S.japonicum Chinese-strain(Hunan-strain,Sjc-H)and S.mansoni HGPRT,respectively.The results of SDS-PAGE and Western blot revealed that the molecular weight of expressed protein was around 30 kDa and could be recognized by anti-His-G-HPR antibody and sera from mice and human with schistosomasis japonica.Conclusion The recombinant plasmid containing SjcHGPRT cDNA is successfully constructed and its expression protein(reSjcHGPRT)is also successfully purified.