This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

Endometriosis(EMs)is a common estrogen-depend-ent clinical disease with the pathological characteristics of malig-nant tumors,which has great impact on women's physical and mental health.In recent years,experimental exploration has re-vealed that ectopic foci are in a hypoxic environment outside the uterine cavity,and mitochondria,as the"functional factories"of the cells,play an important role in the process of planting and in-vasion,and the mitochondrial quality control system,which in-cludes mitochondrial oxidative stress,kinetics,autophagy,bio-genesis and calcium homeostasis,is a key mechanism for the e-quilibrium of the mitochondrial function.The mitochondrial quality control system,including mitochondrial oxidative stress kinetics,autophagy,biogenesis and calcium homeostasis,is a key mechanism for mitochondrial functional balance.Therefore,to clarify the role of the mitochondrial quality control system in the development of EMs with the help of rational and rigorous experi-mental and clinical studies can not only help to clarify the patho-genesis of the disease,but also explore the key targets in the prevention and treatment of the disease.Therefore,this article summarizes the research progress of mitochondrial quality control system in endometriosis,with a view to providing reference and theoretical basis for the etiology,pathogenesis and prevention strategies of EMs.

A second-order calibration method combined with excitation-emission matrix(EEM)fluorescence spectroscopy was presented for simultaneous quantitative analysis of two anti-hypertensive drugs,metolazone(MET)and valsartan(VAL),in human serum and urine,and the quantitative results were compared with the results obtained by zero-and first-order calibration methods.The results indicated that the methods based on zero-and first-order calibration were inadequate for accurately quantifying the components of interest in cases where severe spectral overlap and unknown interferences coexisted.However,it was possible to obtain satisfactory results with the second-order calibration method based on alternating normalization-weighted error(ANWE)algorithm because of its strong"mathematical separation",even when the fluorescence spectra of the target analytes and unknown interferents considerably overlapped.Correlation coefficients for both analytes were greater than 0.99,with mean recoveries of 104.9%±5.7%and 107.8%±9.2%for MET and VAL in human serum,and 103.7%±8.9%and 94.7%±3.8%in human urine,respectively.In addition,the sensitivity,selectivity,limit of detection,limit of quantification,repeatability,and reproducibility of the proposed second-order calibration method were thoroughly examined.All results indicated that the established method was capable of achieving simultaneous and accurate quantification of MET and VAL in human body fluids,which was expected to be applied to analysis of both drugs in clinical settings.

Objective:To investigate the clinical effect of chimeric flap with perforating branch of superficial circumflex iliac artery in reconstruction of complex tissue defect in Achilles tendon region.Methods:A retrospective analysis was conducted on 9 patients who underwent reconstruction of composite tissue defects in Achilles tendon region with the perforator flap of lateral femoral circumflex artery, from January 2017 to January 2023 in the Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University. The patients were 7 males and 2 females and at 16 to 54 years, with 36.6 years in average. Causes of injury were: 5 patients by traffic accident, 2 by machine strangulation and 1 by heavy object, also 1 defect was caused by infection. The soft tissue defects were 4.5 cm×6.0 cm to 8.0 cm×12.0 cm in size, and the defects of Achilles tendon were 4.5 cm to 11.5 cm, with an average of 6.8 cm in length. The wounds were managed with VSD after primary surgery, and then stage Ⅰ surgery were followed to reconstruct the defects in Achilles tendon region with perforator flap of lateral femoral circumflex artery. The sizes of chimeric flaps were 6.5 cm×5.0 cm to 18.0 cm×6.5 cm, including 3 lobuated flaps. The fascia grafts were 5.5 cm to 12.0 cm, with an average of 7.0 cm. Scheduled postoperative follow-ups were conducted in outpatient clinic and via telephone or WeChat interviews, together with the advices for rehabilitation training. Postoperative functions of the Achilles tendon and ankle function were evaluated using Arner-Lindholm criteria and the ankle and hindfoot functional score of American Orthopaedic Foot and Ankle Society (AOFAS).Results:All of the 9 chimeric flaps survived and all the donor sites had primary healing. All patients were entered into the postoperative follow-up for 10 to 66 months, with 14.6 months in average. One chimeric flap was found bulky and a flap thinning surgery was further carried out in 3 months under the request of patient. The rest of patients were satisfied with the appearance and texture of the flaps. The range of ankle plantar flexions was found at 42.9°±1.8° and the dorsal flexions was at 16.2°±2.5°. There was no significant decrease in circumference and no discomfort or muscle re-rupture in the affected legs. Thompson test was found negative. There was no wound related complications during follow-up. Reconstructive surgery were found effective based on Arner-Lindholm criteria, with excellent in 6 patients, good in 2 and 1 in fair. The ankle and subtalar joint function was evaluated according to the ankle-hindfoot function score of AOFAS, with 7 patients scored 90-100 point and 2 scored 75-89 point, and 7 patients were in excellent and 2 in good.Conclusion:The chimeric perforator flap of lateral femoral circumflex artery is safe and reliable in reconstruction of composite tissue defects in Achilles tendon region. It can effectively restore the function and appearance of ankle and hindfoot in patients with composite tissue defects in the Achilles tendon region.

Objective:To explore the clinical effect of thin superficial circumflex iliac artery perforator flap with venous superdrainage in treatment of wound in extremities.Methods:Clinical data of 20 patients who were treated from January 2018 to January 2024 in the Department of Burns and Plastic Surgery, Affiliated Hospital of Zunyi Medical University were retrospectively analysed. The soft tissue defects in extremity were reconstructed with thin perforator flaps of superficial iliac circumflex artery with venous superdrainage. There were 13 patients with upper limb defects and 7 with ankle defects. Of the defects, 12 were of trauma, 2 of tumour resection, 3 of scar release surgery and 3 of donor flat harvesting. The defects ranged from 5.0 cm×4.0 cm to 18.0 cm×7.0 cm in size. During the surgery, portable Doppler was used to detect the perforating branches of the superficial rotational iliac artery and design flaps. The flaps were 5.0 cm×5.0 cm to 20.0 cm×7.0 cm in size, including 16 single-lobed flaps, 3 double-lobed flaps and 1 triple-lobed flaps. The flaps were 2-7 mm in thickness, with an average of 4 mm. Based on the distribution of perforators, the flaps carried the superficial helioiliac artery and the accompanying vein, as well as the superficial helioiliac vein. The superficial helioiliac artery and the accompanying vein were anastomosed during the surgery, and the superficial helioiliac vein was anastomosed to the superficial or deep vein of the recipient site for superreflux. The donor sites were all directly sutured and closed. Postoperative follow-up was conducted by visits of outpatient clinic, and via telephone and WeChat interviews. The survival and appearance of the flaps and complications were observed.Results:A total of 22 arteries and 42 veins of the 20 flaps were anastomosed in surgery. All flaps survived. The donor sites were closed in the first stage. No vascular compromise occurred. One patient had early exudate under the flap on the exposed wound of interphalangeal joint, which healed after routine dressing change and drainage. All patients were included in the postoperative follow-up, with a peroid over 5 to 40 months, at 10.8 months in average. The flaps were thin and soft, with good wear resistance and without pigmentation. The healing of donor sites was good except 1 donore site that had early lymphatic leakage, which was cured after compression and drainage. A linear scar left at the donor sites and it was acceptable to the patients.Conclusion:The treatment of defective soft tissue wounds in extremities using thin perforator flap with venous superdrainage of the superficial circumflex iliac artery is safe and feasible. No further flap thinning surgery is required, and there is a reliable clinical effect.

Aim To explore the mechanism of Jiawei Shaofu Zhuyu decoction in antagonizing endometriosis fibrosis by regulating mitophagy.Methods After the animal model was constructed,the syndrome was evalu-ated by general condition,organ water content and ther-mal imaging.The curative effect was evaluated by the weight of ectopic focus and the degree of adhesion.The pathological changes were compared using HE stai-ning,transmission electron microscopy,Masson and Sir-ius red staining.The expression of PINK1 and Parkin was detected by immunohistochemistry.The expression of mRNA and protein was determined by qPCR and Western blot,and the level of serum ROS was detected by ELISA.Results The autonomic activity of model mice was weakened,the water content of organs rose,and the temperature of limbs and lower abdomen was reduced by thermal imaging.HE staining showed obvi-ous hyperplasia of ectopic epithelium and glands.Transmission electron microscopy showed mitochondrial and endoplasmic reticulum structure damage,and nor-mal autophagy structure disappeared.Masson and Siri-us red staining showed increased collagen deposition;immunohistochemistry showed decreased expression of PINK1 and Parkin in ectopic foci.qPCR and Western blot showed that the expression of PINK1,Parkin,Bec-lin1,LC3 mRNA and protein in ectopic foci of model mice decreased,the expression of p62 mRNA and pro-tein increased,and serum ROS increased.The syn-drome performance of model mice was improved after the intervention of Jiawei Shaofu Zhuyu decoction;the inflammatory infiltration of ectopic foci was relieved,the morphology of mitochondria and endoplasmic retic-ulum was restored,and normal autophagy structure ap-peared.The degree of collagen deposition and fibrosis was reduced;the mRNA and protein expression of PINK1,Parkin,Beclin1 and LC3 increased.The ex-pression of p62 mRNA and protein decreased,and the level of ROS decreased.Conclusions Jiawei Shaofu Zhuyu decoction can improve the fibrosis of ectopic le-sions in mice with endometriosis of cold-dampness sta-sis syndrome,which may be related to the regulation of mitophagy.

Objective:To explore the clinical application value of infrared thermal imaging in assisting the repair of complex joint tissue defects in children using the lateral femoral circumflex artery (LFCA) chimeric perforator flap.Methods:A retrospective analysis was conducted on clinical data from the Department of Burns and Plastic Surgery at the Affiliated Hospital of Zunyi Medical University between May 2019 and September 2023. The study included cases where the LFCA perforator chimeric flap was used to repair complex joint tissue defects in children. Preoperative perforator mapping was performed using an infrared thermal imaging device combined with a portable Doppler. The flap and fascial flap were designed based on the distribution of hot spots and thermal lines. Intraoperative confirmation of perforator distribution was carried out. Postoperative follow-ups were conducted to assess flap survival, the need for secondary thinning procedures, wound healing status, donor site scarring, joint swelling, and functional recovery.Results:A total of 11 pediatric patients were enrolled, including 8 males and 3 females, aged 2 to 14 years (mean age: 5.8 years). The injury mechanisms included traffic accidents (6 cases), wheel spoke injuries (2 cases), falls (2 cases), and heavy object crush injuries (1 case). Affected joints included the wrist (1 case), knee (2 cases), and ankle (8 cases). Associated injuries included metacarpal fracture (1 case), metatarsal fractures (2 cases), and extensor tendon defects (1 case). Tendon defect length was 3 cm, joint capsule defect area ranged from 2 cm × 3 cm to 4 cm × 6 cm, and fascial flap size ranged from 2 cm × 4 cm to 2 cm × 14 cm. Skin defect area ranged from 5 cm × 3 cm to 16 cm × 9 cm, and flap size ranged from 6 cm × 4 cm to 15 cm × 5 cm. Preoperative infrared thermography identified 28 hot spots, with 26 perforators confirmed intraoperatively (positive rate: 92.9%). Thermal line distribution zones consistently corresponded to perforator branches. All 11 flaps survived, with primary healing achieved at both donor and recipient sites. Follow-up duration ranged from 6 to 36 months (mean: 10.5 months). All recipient sites exhibited soft flap texture. One case required secondary debulking due to flap bulkiness. Donor sites showed only linear scarring. During follow-up, no significant joint swelling, functional impairment, or joint dislocation was observed.Conclusion:Infrared thermal imaging-assisted perforator mapping for the design of LFCA chimeric perforator flaps is a safe and reliable method for repairing complex joint tissue defects in children. It provides accurate perforator localization, offering significant clinical value for the design and harvesting of LFCA chimeric perforator flaps in pediatric patients.

Objective To investigate the association of endometrial thickness(EMT)with obstetric and neonatal outcomes of monoparous pregnancy in fresh cleavage embryos transfer.Methods A total of 1 845 patients of monoparous pregnancy after fresh cleavage embryos transfer cycles from Jan 2016 to Mar 2022 at Shanghai First Maternity and Infant Hospital,Tongji Universtiy were analyzed retrospectively.Patients were categorized into three groups by EMT on transferation day:≤8 mm(group A),8-14 mm(group B)and≥14 mm(group C).The primary outcomes were preterm birth(PTB),birth weight and birth weight z-score,small-for-gestation age,large-for-gestation age,very low birth weight,low birth weight and macrosomia.The second outcomes were pregnancy and perinatal complications.The relationship between EMT and adverse neonatal outcomes was estimated by Logistic regression analysis.Results The rate of ectopic pregnancy was increased significantly in group A.No significant differences were found among the three groups in gestation age,birth weight,birth weight z-score,PTB,small for gestation age,large for gestation age,low birth weight,very low birth weight and macrosomia.Compared with group B,the odds of adverse neonatal outcomes did not show significant differences before and after adjustment in both group A and group C by Logistic regression analysis.Conclusion Thinner EMT in fresh cleavage embryos transfer is associated with higher rate of ectopic pregnancy,while it is not independently associated with adverse perinatal outcomes.

Objective To investigate the association of endometrial thickness(EMT)with obstetric and neonatal outcomes of monoparous pregnancy in fresh cleavage embryos transfer.Methods A total of 1 845 patients of monoparous pregnancy after fresh cleavage embryos transfer cycles from Jan 2016 to Mar 2022 at Shanghai First Maternity and Infant Hospital,Tongji Universtiy were analyzed retrospectively.Patients were categorized into three groups by EMT on transferation day:≤8 mm(group A),8-14 mm(group B)and≥14 mm(group C).The primary outcomes were preterm birth(PTB),birth weight and birth weight z-score,small-for-gestation age,large-for-gestation age,very low birth weight,low birth weight and macrosomia.The second outcomes were pregnancy and perinatal complications.The relationship between EMT and adverse neonatal outcomes was estimated by Logistic regression analysis.Results The rate of ectopic pregnancy was increased significantly in group A.No significant differences were found among the three groups in gestation age,birth weight,birth weight z-score,PTB,small for gestation age,large for gestation age,low birth weight,very low birth weight and macrosomia.Compared with group B,the odds of adverse neonatal outcomes did not show significant differences before and after adjustment in both group A and group C by Logistic regression analysis.Conclusion Thinner EMT in fresh cleavage embryos transfer is associated with higher rate of ectopic pregnancy,while it is not independently associated with adverse perinatal outcomes.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.