Objective:To investigate the clinical effect of anterolateral thigh perforator flap (ALTPF) combined with transfer of fascia lata in reconstruction of complex tissue defects of hand and foot.Methods:From July 2021 to October 2023, 9 patients with complex tissue defects of hand and foot were treated with ALTPF combined with fascia lata in the Department of Orthopaedic Microsurgical Repair and Reconstruction of Fuyang People's Hospital Affiliated to Anhui Medical University. There were 2 males and 7 females with an average age of 28.1 (range, 4-65) years old. Three patients had extensor tendon defects in 6 digits of dorsal hands, 5 had extensor tendon defect in 10 toes of dorsal foot, 1 had a defect of anterior tibial tendon and 1 had Achilles tendon defect in posterior ankles. The sizes of soft tissue defect ranged from 8.0 cm×6.0 cm to 15.0 cm×10.0 cm, and the lengths of tendon defect ranged from 6.0 to 13.0 cm. Preoperative Doppler ultrasound was used to locate the distribution of perforating branches in the anterolateral thigh region. According to the characteristics of wound, ALTPFs and fascia lata were designed and harvested. Fascia lata with an appropriate size of 1.5 cm×8.0 cm-4.5 cm×15.0 cm were taken to bridge the defects of the tendon and the Achilles tendon. The wounds were reconstructed with flaps sized 9.0 cm×6.5 cm-18.0 cm×7.5 cm. Nine fascia lata donor sites and 8 flap donor sites were sutured directly. One donor site was treated with a skin graft of ipsilateral ilioinguinal region. The survival and complications of the flaps and donor sites were observed through outpatient follow-up visits, WeChat reviews and home visits, etc. The hand function was assessed according to the Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association, and the foot and ankle function was assessed according to the Mazur score standard of joint range of motion and motor function.Results:All patients were included in follow-up for 4-24 (mean, 13.4) months with complete clinical data being collected. All 9 ALTPFs survived and healed primarily. A linear scar left in donor sites in 8 patients, and mild lamellar scar at skin graft in 1 patient. Texture and colour of the flaps were similar to the surrounding tissue without secondary flap thinning surgery. Combined with postoperative rehabilitation training, satisfactory function recoveries were achieved. Hand function of 3 patients were evaluated according to Evaluation Standard of Upper Limb Functional of Hand Surgery of Chinese Medical Association, 2 patients were excellent and 1 was good. Ankle and foot functions in 6 patients were evaluated according to the range of motion of ankle and foot and Mazur score standard for motor function, 4 patients were excellent and 2 were good.Conclusion:ALTPF combined with fascia lata transfer can reconstruct the complex tissue defects of hand and foot. Of which, 1 donor site can meet the requirements of 2 types of tissues reconstruction at the same time, and with minimal damage to the donor site as well as an precise reconstruction of the recipient site. It avoids staged surgery, shortens the treatment time and reduces the cost of treatment.

In order to reveal the intestinal absorption mechanism of saikosaponin d (SSd) in vitro and in vivo, the current research investigated the effects of different experimental conditions (time, concentration, temperature, pH, intestinal segments), transporter inhibitors, paracellular pathway enhancer, metabolic enzyme inhibitors on the intestinal absorption of SSd, in Caco-2 monolayers and a single pass perfusion model in rats.The results showed that the apparent permeability coefficient (Papp) and effective permeability coefficient (Peff) of SSd were 4.75 × 10-7 - 6.38 × 10-7 cm/s and 0.19 × 10-4- 0.27 × 10-4 cm/s, respectively, indicating that it was a low permeability compound, and that the transmembrane transport of SSd was concentration-dependent (0.5-5 μmol/L) and time-dependent (0-180 min).Ileum was the main absorption site for SSd. Experimental results based on Caco-2 monolayers showed that the P-gp inhibitor and paracellular permeability enhancer significantly increased the absorption of SSd (P < 0.05), which was consistent with the results obtained in rats. Inhibitors of OATPs and OCTs showed different results in vitro and in vivo, which may be related to the lower expression of them in jejunum.In summary, the intestinal absorption of SSd occurs through a carrier-mediated and energy-dependent transport, as well as passive diffusion, and P-glycoprotein plays an important role in the active transport of SSd.

Objective:To investigate the relationship between the positive surgical margin and clinical factors such as neoadjuvant hormonal therapy after robot-assisted laparoscopic radical prostatectomy (RARP) in high-risk patients with prostate cancer.Methods:The clinical data of 164 patients with high-risk prostate cancer being performed RARP by one surgeon were analyzed retrospectively in our hospital from January 2016 to January 2022. The mean patient’s age was (65.3±6.2) years old, mean body mass index (BMI) was (25.6±3.0) kg/m 2, the median value of total prostate specific antigen (tPSA) before operation was 18.6(11.3, 31.3)ng/ml, the median value of Gleason score before operation was 7 (7, 8), the median value of prostate volume was 29.3 (22.4, 40.2) ml, and the clinical stage was T 2aN 0M 0-T 4N 0M 0. 80 patients with prostate cancer were treated with neoadjuvant endocrine therapy. All of them were treated with complete androgen blockade with a median course of 3 months. Univariate analysis was used to analyze the correlation between age, BMI, prostate volume, neoadjuvant hormonal therapy, preoperative tPSA, clinical stage, Gleason score before operation and positive surgical margin. Then multivariate logistic regression was used to further analyze the independent risk factor of positive surgical margin after RARP. Results:The postoperative pathological diagnosis included pT 2 stage in 111 cases (67.7%), pT 3a stage in 15 cases (9.1%), pT 3b stage in 25 cases (15.2%), pT 4 stage in 13 cases (7.9%). No lymph node metastasis was noticed in all patients. The Gleason scores included 6 in 11 cases (6.7%), 3+ 4 in 26 cases (15.9%), 4+ 3 in 36 cases (22.0%), 8 in 17 cases (10.4%), 9-10 in 24 cases (14.6%), un-evaluation due to endocrine therapy in 50 (30.5%). The positive surgical margin of high-risk patients with prostate cancer was 44.5% (73/164). Univariate analysis showed that the neoadjuvant hormonal therapy, tPSA and clinical stage were correlated with positive surgical margin ( P<0.05). Multivariate logistic regression analysis showed that non-neoadjuvant hormonal therapy, preoperative tPSA>20ng/ml and clinical stage>T 2b were independent risk factors for positive surgical margin of high-risk patients with prostate cancer. Stratified analysis showed that when the preoperative tPSA was 10-20 ng/ml(21.1% vs.55.9%, P=0.014), the clinical stage was T 2c(29.6% vs.49.1%, P=0.040), the Gleason score before operation was 7(19.4% vs.54.1%, P=0.003), the positive surgical margin of high-risk patients in the neoadjuvant hormonal therapy group was significantly lower than that in the non-neoadjuvant hormonal therapy group ( P<0.05). Conclusions:Non-neoadjuvant hormonal therapy, preoperative tPSA>20 ng/ml and clinical stage>T 2b were independent risk factors for positive surgical margin of RARP in the high-risk patients with prostate cancer. For high-risk patients with preoperative tPSA of 10-20 ng/ml, clinical stage of T 2c and Gleason score before operation of 7, neoadjuvant hormonal therapy has important clinical significance in reducing the positive surgical margin of RARP.

Objective To explore the role and mechanism of HMGB1 in the fatty acid metabolism reprogramming and mitochondrial fusion/fission of hypoxic and nutrient-poor pancreatic cancer cells. Methods The correlation between the expression level of HMGB1 in pancreatic cancer tissue and the survival rate of pancreatic cancer patients were analyzed by GEPIA database. CCK-8 assay was used to measure cell proliferation rate, and scratch test and Transwell chamber method were carried out to detect the effects of endogenous HMGB1 on the invasion and migration abilities of human pancreatic cancer cell line Patu8988 after hypoxic and nutrient-poor treatment. Laser confocal microscope was used to observe the changes of mitochondrial morphology of Patu8988 cells. Western blot was used to detect the expression levels of mitochondrial fusion/fission and de novo fatty acid synthesis-related proteins. Results GEPIA database analysis results showed that HMGB1 was highly expressed in pancreatic cancer tissues (P < 0.01), and the expression level was negatively correlated with the survival time of pancreatic cancer patients (P=0.00097). Knockdown of HMGB1 expression could inhibit the proliferation, invasion and migration abilities of Patu8988 cells under hypoxic and nutrient-poor conditions. However, mitochondrial fission in patu8988 cells was increased. Knockdown of HMGB1 in Patu8988 cells increased the expression of fission-related protein FIS1 while decreased the expression of p-DRP1(Ser637) and fusion-related protein MFN1 and MFN2 in hypoxic and nutrient-poor environment; ACLY, p-ACLY and FASN protein expression levels were down-regulated. Conclusion Endogenous HMGB1 can promote the fusion and inhibit the fission of mitochondria in hypoxic and nutrient-poor Patu8988 cells, maintain mitochondrial morphology and function, and thereby up-regulate ACLY protein expression and phosphorylation level, promote FA synthesis, and maintain the proliferation, invasion and migration abilities of pancreatic cancer cells.

Saikosaponins (SSs) are the main active components extracted from Bupleuri Radix (BR) which has been used as an important herbal drug in Asian countries for thousands of years.It has been reported that the intestinal bacteria plays an important role in the in vivo disposal of oral SSs.Although the deglycosylated derivatives (saikogenins,SGs) of SSs metabolized by the intestinal bacteria are speculated to be the main components absorbed into the blood after oral administration of SSs,no studies have been reported on the characteristics of SGs for their intestinal absorption,and those for SSs are also limited.Therefore,a rapid UHPLC-MS/MS method was developed to investigate and compare the apparent permeability of three common SSs (SSa,SSd,SSb2) and their corresponding SGs (SGF,SGG,SGD) through a bidirectional transport experiment on Caco-2 cell monolayer model.The method was validated according to the latest FDA guidelines and applied to quantify the six analytes in transport medium samples extracted via liquid-liquid extraction (LLE).The apparent permeability coefficient (Papp) determined in this study indicated that the permeability of SGs improved to the moderate class compared to the corresponding parent compounds,predicting a higher in vivo absorption.Moreover,the efflux ratio (ER) value demonstrated an active uptake of SSd and the three SGs,while a passive diffusion of SSa and SSb2.

Objective Reference standard of the RPOB(rifampin resistance)gene recommended by CLSI-MM18A(Interpretive Criteria for Identification of Bacteria and Fungi by DNA Target Sequencing) was used to evaluate the ability of MALDI-TOFMS techniques for the identification and classification of non-tuberculous Mycobacterium.Methods Fifty five clinicalstrains were collected from 2012 to 2016 with different sources.The RPOB gene was sequenced, and results were applied to phylogenetics analysis. MALDI-TOF MS technology was implemented to identify the strains, and cluster analysis was conducted based on protein fingerprint.The consistency of two methods for NTM identification and typing was evaluated.Results The RPOB gene method showed a good ability of identification(similarity>99.0%) and subtyping(to subspeciesof the complex level).The French BioMérieux MALDI-TOF MS identified 89.1% of 55 strains to genus level and 78.2% to species level.The phylogeneticsanalysis of protein fingerprint by SARAMS Premium software also showed good typing ability.Conclusions MALDI-TOF MS technology can identify and classify non-tuberculous Mycobacterium effectively,which is rapid and easy.It is complementary to RPOB gene method in laboratory application.

Objective: Establishing the mass spectrum library of a new Campylobacter- " C.fetus subsp.testudinum" for rapid species identification in clinical microbiology laboratory. Methods: Illumina second generation sequencing platform 2000/miSeq was used to carry out high flux genome sequencing for the strains which were collected to establish mass spectrum library.The analysis oforthologous average nucleotide identity (OrthoANI) between collected strains and reference strains was performed at JAVA 8 operation environment. Then, the mass spectrums ofcollected strains andreference strains were acquired using MALDI-TOF MS. And the mass spectrum library of C. fetus subsp.testudinum. were established and verified. Results: The OrthoANI analysis showed that the OrthoANI value of the collected strains and the reference strain C. fetus subsp.testudinum03-427 was 99.30%-99.96%, while the OrthoANI values of collected strains and C. fetus subsp.venerealisNCTC10354 orC.fetus subsp.fetus82-40 were 91.05%-92.26%. With reference to OrthoANI ≥ 95% as the basis for the determination of the same strain, the strains which collected to establish mass spectrum library was finally identified as " C. fetus subsp.testudinum" . The identification accuracy rate of the mass spectrum library was 100% (consistent with gene sequencing), and the confidence interval was 82.3%-99.9%, identification of the same strain is 100% reproducible. Conclusions The new" gold standard" based on high throughput sequencing and total genome analysis has provided the ideal reference value for the establishment of mass spectrum library.And the accurate and objective reference spectrum of the" C.fetus subsp.testudinum" provides a new platform for the rapid diagnosis of fetal Campylobacter infection. (Chin J Lab Med, 2018, 41: 583-588)

Objective To investigate the early diagnostic ability of conventional imaging methods in micro-hepatocellular carcinoma(M HCC) in order to improve the imaging diagnostic procedure for M HCC.Methods The data of 40 cases of M HCC(52 lesions)and 21 cases of regenerative nodules(35 lesions)confirmed by clinical results were collected continuously.All the patients were examined with ultrasonography (US),plain CT combined with multi-phase enhancement scans,and plain MRI combined with diffusion weighted imaging(DWI).To compare the diagnostic efficacy of three different imaging methods,the imaging data were analyzed retrospectively by χ2test and receiver operating characteristic curve(ROC).Results There was a significant difference in the M HCC detection rate between US and CT,and between US and MRI,respectively(P<0.05).There was no significant difference in the MHCC detection rate between CT and MRI(P>0.05).The sensitivity of US,CT and MRI in the detection of MHCC was 61.5%,80.8% and 92.3%,specificity was 88.6%,91.4% and 97.1%,positive predictive value was 88.9%,93.3% and 98.0%,and area under the ROC curve was 0.751, 0.861 and 0.947,respectively.Plain MRI combined with DWI had the highest diagnostic efficacy,among the three imaging methods. Conclusion US can be used as a primary screening method for M HCC in high risk population,and plain CT combined with multi-phase enhancement scans can be the first choice for early diagnosis of MHCC.For highly suspected the cases with positive AFP but negative findings on US and CT,plain MRI combined with DWI are recommended to rule out the possibility.

Objective To observe the effects of Chinese herbal compounds (alkaloid of Coptidis Rhizoma 40%, total saponin of panax notoginseng 35%, and polysaccharide of Ophiopogonis Radix 25%) on levels of blood glucose, fructosamine, and insulin of diabetic rats, and explore mechanism of action.Methods Ten rats were randomly chosen as blank control group, and other rats were used to establish diabetic rat models by STZ intraperitoneal injection. Successfully molding rats were randomly divided into Chinese herbal compounds high, medium, low dose groups, metformin group, and model group, 10 rats in each group. Each rat received gavage for continuous 4 weeks. Then rat blood was collected by cutting head and taking caudal vein. FBG was detected by glucose oxidase method;insulin level was detected by radio-immunity double antibody method;fructosamine content was detected by fructosamine method.Results Compared with the blank control group, model group the levels of FBG, froctosamine content increased, while fasting insulin level were lower after 4 weeks'medication. Compared with the model group, the levels of FBG, froctosamine content in Chinese herbal compounds high and medium dose groups and metformin group were lower, while fasting insulin level increased after 4 weeks'medication (P<0.05).Conclusion TCM herbal compounds can increased the basic insulin level by decreasing the contents of FBG and fructosamine.

There are lack of specific treatment for diabetic peripheral neuropathy (DPN)right now.However, TCM intervention highlights the distinct advantage for the disease.Modified Huangqi-Guizhiwuwu Tang is one of the important recipes for that.But there are no uniform standards curative effect evaluation for it, and some clinical trials do not have some new evaluation standards.We summary the clinical research literatures in recent years and analyze the existing problems, therefore, provide new recommendations as views for medical researchers in the design of clinical trials.