[Clinical value of the MeltPro MTB assays in detection of drug-resistant tuberculosis in paraffin-embedded tissues].

Objective: To evaluate the clinical value of the MeltPro MTB assays in the diagnosis of drug-resistant tuberculosis. Methods: A cross-sectional study design was used to retrospectively collect all 4 551 patients with confirmed tuberculosis between January 2018 and December 2019 at Beijing Chest Hospital, Capital Medical University. Phenotypic drug sensitivity test and GeneXpert MTB/RIF (hereafter referred to as "Xpert") assay were used as gold standards to analyze the accuracy of the probe melting curve method. The clinical value of this technique was also evaluated as a complementary method to conventional assays of drug resistance to increase the detective rate of drug-resistant tuberculosis. Results: By taking the phenotypic drug susceptibility test as the gold standard, the sensitivity of the MeltPro MTB assays to detect resistance to rifampicin, isoniazid, ethambutol and fluoroquinolone was 14/15, 95.7%(22/23), 2/4 and 8/9,respectively; and the specificity was 92.0%(115/125), 93.2%(109/117), 90.4%(123/136) and 93.9%(123/131),respectively; the overall concordance rate was 92.1%(95%CI:89.6%-94.1%),and the Kappa value of the consistency test was 0.63(95%CI:0.55-0.72).By taking the Xpert test results as the reference, the sensitivity of this technology to the detection of rifampicin resistance was 93.6%(44/47), the specificity was100%(310/310), the concordance rate was 99.2%(95%CI:97.6%-99.7%), and the Kappa value of the consistency test was 0.96(95%CI:0.93-0.99). The MeltPro MTB assays had been used in 4 551 confirmed patients; the proportion of patients who obtained effective drug resistance results increased from 83.3% to 87.8%(P<0.01); and detection rate of rifampicin, isoniazid, ethambutol, fluoroquinolone resistance, multidrug and pre-extensive drug resistance cases were increased by 3.2%, 14.7%, 22.2%, 13.7%, 11.2% and 12.5%, respectively. Conclusion: The MeltPro MTB assays show satisfactory accuracy in the diagnosis of drug-resistant tuberculosis. This molecular pathological test is an effective complementary method in improving test positivity of drug-resistant tuberculosis.

[Detection of tuberculosis genes associated with drug-resistance in paraffin-embedded tissue specimens using next generation sequencing technology].

Objective: To evaluate the use of multiplex PCR amplicon sequencing (mPCR-NGS) technology in detecting gene mutations related to drug resistance of Mycobacterium tuberculosis (MTB) in formalin-fixed paraffin-embedded tissue specimens, and to explore its clinical value in the diagnosis of drug-resistant tuberculosis. Methods: Fifty clinical MTB strains isolated in the Changping District Tuberculosis Control Institute of Beijing from April 2013 to October 2015 with drug susceptibility test (DST) results of rifampicin, isoniazid, ethambutol, streptomycin, ofloxacin, capreomycin, kanamycin and amikacin available were recovered, including 42 drug-resistant strains and 8 drug-sensitive strains. The mPCR-NGS test was established to detect genes related to the 8 anti-tuberculosis drugs according to the previously published studies and databases. Fifty-five paraffin-embedded tissue specimens from drug-resistant tuberculosis patients were collected in the Department of Pathology, Beijing Chest Hospital, Capital Medical University during November 2017 to September 2018. All the specimens showed no less than one mutation in the gene regions related to drug resistance of any of the 4 drugs (rifampicin, isoniazid, ethambutol or fluoroquinolones) by probe melting curve assay. The effectiveness of mPCR-NGS test was evaluated on clinical MTB isolates using phenotypic DST as the reference. Clinical evaluation of mPCR-NGS test on formalin-fixed paraffin-embedded specimens from TB patients was performed using probe melting curve assay as the reference. The sensitivity, specificity and coincidence of mPCR-NGS were analyzed. Results: Using phenotypic DST as the reference, the sensitivities of the mPCR-NGS for detecting drug-resistance of rifampicin, isoniazid, streptomycin, and ethambutol were 95% (38/40), 93% (27/29), 93% (27/29), and 72% (13/18), respectively; and the specificities were 100% (10/10), 95% (20/21), 100% (21/21), and 94% (30/32), respectively. The sensitivities for capreomycin, kanamycin and amikacin were all 100% (2/2, 3/3, 3/3), and the specificities were 98% (47/48), 100% (33/33) and 100% (47/47), respectively. The sensitivity and specificity of ofloxacin were 70% (7/10) and 100% (40/40), respectively. The total coincidence rate for the 8vdrugs was 94%, and the Kappa value was 0.87. The 55 paraffin-embedded tissue specimens included in this study were all tested by probe melting curve assays. Among them 28 were resistant to rifampicin, 37 resistant to isoniazid, 13 resistant to ethambutol, and 17 resistant to fluoroquinolones. Using the probe melting curve assay as the reference, the sensitivities of the mPCR-NGS for detecting resistant to rifampicin, isoniazid, ethambutol, and fluoroquinolones were 100% (28/28), 95% (35/37), 100%, and 100%, respectively; and the specificities were all 100% (42/42, 38/38). The total coincidence rate of the two methods was 99%, and the Kappa value was 0.98. Conclusions: mPCR-NGS showed good sensitivities and specificities in detecting drug-resistant gene mutations both in clinical MTB isolates and paraffin-embedded tissue specimens. mPCR-NGS has the potential to be an accurate and rapid molecular pathological technology for diagnosis of drug-resistant tuberculosis.

[Association between platelet-to-lymphocyte ratio and neutrophil-to-lymphocyte ratio with disease activity in Takayasu arteritis patients].

Objective: To explore the relation between platelet-to-lymphocyte ratio (PLR) or neutrophil-to-lymphocyte ratio (NLR) with disease activity in Takayasu arteritis (TA) patients. Methods: Present retrospective study involved 289 patients with TA, who were hospitalized in our department between January 2010 and October 2017, and 280 age and gender matched healthy controls,who underwent thealth examination in our health examination center during the same period (control group). TA patients were further divided into active and inactive groups (180 and 109 cases respectively) according to Kerr scores. The clinical data were compared between groups. Pearson correlation analysis was used to evaluate the relationship between PLR or NLR and disease activity (Kerr score or C-reactive protein or erythrocyte sedimentation rate). Receiver operating characteristic (ROC) curve was employed to judge the cut-off value of disease activity for TA patients. Results: PLR and NLR were significantly higher in TA group than in control group(137.33 (97.38, 193.37) vs. 120.55 (96.86, 144.60) and 2.38 (1.76, 3.57) vs. 1.66 (1.35, 2.08) , respectively, all P<0.001). PLR and NLR were significantly higher in active TA group than in inactive TA group (163.43 (123.64, 224.15) vs. 110.53 (84.22, 147.24) and 2.59 (1.96, 3.94) vs. 1.95 (1.53, 2.86) respectively, all P<0.001). PLR and NLR of active group were significantly decreased after 6 months treatment (164.05 (123.29, 226.29) vs. 104.67 (77.22, 138.43) and 2.58 (1.96, 3.91) vs. 2.15 (1.67, 2.60) respectively, all P<0.001). PLR was positively correlated with Kerr score (r=0.439, P<0.001), C-reactive protein (r=0.328, P<0.001) and erythrocyte sedimentation rate (r=0.410, P<0.001). NLR also exhibited a positive relationship with Kerr score (r=0.235, P<0.001), C-reactive protein (r=0.169, P=0.005) and erythrocyte sedimentation rate (r=0.123, P=0.037). A PLR level of 176.709 was shown to be the best predictive cut-off value for TA disease activity (sensitivity 44.6%, specificity 93.0%, and area under the curve=0.766).A NLR level of 2.128 was shown to be the best predictive cut-off value for TA disease activity (sensitivity 70.9%, specificity 47.7%, and area under the curve=0.691). Conclusion: PLR and NLR are useful markers for predicting disease activity of TA patients.

Cytogenetic and molecular identification of a wheat-Leymus mollis alien multiple substitution line from octoploid Tritileymus x Triticum durum.

Leymus mollis (Trin.) Pilger (NsNsXmXm, 2n = 28), a wild relative of common wheat, possesses many traits that are potentially valuable for wheat improvement. In order to exploit and utilize the useful genes of L. mollis, we developed a multiple alien substitution line, 10DM50, from the progenies of octoploid Tritileymus M842-16 x Triticum durum cv. D4286. Genomic in situ hybridization analysis of mitosis and meiosis (metaphase I), using labeled total DNA of Psathyrostachys huashanica as probe, showed that the substitution line 10DM50 was a cytogenetically stable alien substitution line with 36 chromosomes from wheat and three pairs of Ns genome chromosomes from L. mollis. Simple sequence repeat analysis showed that the chromosomes 3D, 6D, and 7D were absent in 10DM50. Expressed sequence tag-sequence tagged sites analysis showed that new chromatin from 3Ns, 6Ns, and 7Ns of L. mollis were detected in 10DM50. We deduced that the substitution line 10DM50 was a multiple alien substitution line with the 3D, 6D, and 7D chromosomes replaced by 3Ns, 6Ns, and 7Ns from L. mollis. 10DM50 showed high resistance to leaf rust and significantly improved spike length, spikes per plant, and kernels per spike, which are correlated with higher wheat yield. These results suggest that line 10DM50 could be used as intermediate material for transferring desirable traits from L. mollis into common wheat in breeding programs.

Development of 5Ns chromosome-specific SCAR markers for utilization in future wheat breeding programs.

In previous studies, we developed a wheat-Psathyrostachys huashanica Keng disomic addition line 3-8-10-2, which exhibited high stripe rust resistance and could be used as a donor source for introducing novel disease resistance gene(s) into wheat in future breeding programs. It was identified using cytology, genomic in situ hybridization (GISH), EST-SSR, EST-STS and morphological analyses. However, these techniques are not suitable for breeding programs that require the rapid screening of large numbers of genotypes because they are highly technical and time-consuming. In this study, three Ns genome-specific SCAR markers were developed via random amplified polymorphic DNA (RAPD) markers. These SCAR markers were further validated using a complete set of wheat-P. huashanica disomic addition lines, which segregated the 5Ns disomic addition line individuals. Our results indicated that the SCAR markers associated with the 5Ns chromosome of P. huashanica and they provide a low cost, high efficiency, alternative tool for screening 5Ns chromosomes in a wheat background. These newly developed SCAR markers that species-specificity of the markers was proved by analysis of a wide range of cereal species, and specific for 5Ns chromosome, which should be useful in marker-assisted selection for wheat breeders who want to screen genotypes that may contain 5Ns chromatin.

A novel SCAR marker for detecting Psathyrostachys huashanica Keng chromatin introduced in wheat.

In this study, we cloned and sequenced a 938-base pair polymorphic band, pHs27, in the tightly linked random amplified polymorphic DNA marker OPU10 and converted it into a sequence-characterized amplified region (SCAR) marker referred to as RHS141, which was specific for the Ns genome of Psathyrostachys huashanica. A GenBank basic local alignment search tool search showed that the sequence of pHs27 had no primary sequence homology with known sequences, and Southern blotting confirmed this result. This SCAR marker was used to detect Ns genome chromatin in wheat, and it was successfully amplified in P. huashanica itself, a complete set of wheat-P. huashanica disomic addition lines (1Ns-7Ns), and undetermined homoeologous group addition lines. This SCAR marker will be a powerful tool for the marker-assisted selection of P. huashanica chromosome(s) in a wheat background, and it should also allow wheat breeders to screen for the excellent traits found in P. huashanica chromatin.

[Clinical effect of the giant deep inferior epigastric artery paraumbilical perforator flap in repairing the circular high-voltage electric burn wounds on the wrist].

Objective: To investigate the clinical effect of the giant deep inferior epigastric artery paraumbilical perforator flap in repairing the circular high-voltage electric burn wounds on the wrist. Methods: A retrospective observational study method was used. From September 2016 to October 2021, thirteen male patients (aged 20-43 years) with annular high voltage (10-100 kV) electrical burns on the wrist were admitted to the Beijing Jishuitan Hospital. At the early stage after injury, the patient's wrist was subjected to incision, tension reduction and debridement, with the wound area after debridement being 27 cm×16 cm-32 cm×19 cm; in 12 patients with vascular injury, the radial or ulnar artery was reconstructed by great saphenous vein transplantation, with the length of 15-25 cm; the wrist wound was repaired by free transplantation of the deep inferior epigastric artery paraumbilical perforator flap (if the wound was giant, the lower abdominal flap carrying other perforators was used), with the area of 30 cm×19 cm-35 cm×20 cm. The donor site was repaired by direct suture+skin grafting or relay flap transplantation. After surgery, the survival of flap in recipient area, as well as survival of the skin or flap in donor site were observed. During follow-up, the appearances of the flap in recipient area and the recovery of hand function, as well as the healing of donor site, occurrence of abdominal wall hernia, and scar in skin graft area were observed. Results: After surgery, all the 13 patients' paraumbilical perforator flaps survived. Among them, 3 patients had subcutaneous fat necrosis at the distal end of the wrist flap, and the wound had mild infection, which healed after re-expansion and dressing change. All the skin grafts in the donor site of 10 patients survived, and the flaps in the donor site of 3 patients survived well. The patients were followed up for 6 months to 3 years. The flaps in recipient area were in good shape, 8 cases had partial recovery of hand function, and 5 cases had loss of finger flexion function; the donor site of abdominal flap healed well with no abdominal hernia occurred, and the skin graft site had no obvious scar hyperplasia and was soft in texture. Conclusions: Early vascular reconstruction after injury, together with free transplantation of the giant deep inferior epigastric artery paraumbilical perforator flap are effective in repairing circular high-voltage electrical burn wounds on the wrist.

[Exploration on clinical treatment of incisional wound infection after orthopedic internal fixation for lower extremity joint injuries].

Objective: To explore the clinical treatment of incisional wound infection after orthopedic internal fixation for lower extremity joint injuries. Methods: A retrospective cohort study was conducted. From February 2014 to December 2019, 214 patients with lower limb closed injury were treated in Beijing Jishuitan Hospital, including 143 males and 71 females, aged from 16 to 65 years. All patients underwent orthopedic internal fixation, after which incisional wound infection developed in 42 cases of postoperative wounds of patellar fracture, 30 cases of postoperative wounds of tibial plateau fracture, 72 cases of postoperative wounds of Achilles tendon rupture, 45 cases of postoperative wounds of calcaneal fracture, and 25 cases of postoperative wounds of Pilon fracture with 31 cases of superficial infection and 183 cases of deep infection. According to the postoperative evaluation of the incisional wounds, dressing change and/or thorough debridement (with wound area from 4 cm×3 cm to 11 cm×5 cm after debridement), and internal fixation treatment were performed. After thorough debridement, wounds were treated with continuous vacuum sealing drainage (VSD), and then direct suture+VSD or flap transplantation were performed according to the incision and its surrounding skin tissue, infection, suture tension, exposure of internal fixation or bone or Achilles tendon tissue. According to the wound site and injury, the flap types of retrograde anterolateral thigh perforator flap, medial sural artery perforator flap, gastrocnemius myocutaneous flap, sural neurovascular flap, peroneal brevis muscle flap, posterior tibial artery perforator flap, and free anterolateral thigh perforator flap could be selected. The donor site wound was closed by direct suture or grafted with split-thickness skin graft. The removal of internal fixation, wound repair method, type and size of flaps, survival of flaps, and wound healing were recorded. The recurrence of infection, appearance of donor and recipient areas, and recovery of lower limb joint function were followed up. Results: All the internal fixations were removed in patients with postoperative wounds of patellar fracture, among whom 36 cases were repaired with retrograde anterolateral thigh perforator flap, and 6 cases were repaired with medial sural artery perforator flap. Among the patients with postoperative wounds of tibial plateau fracture, the internal fixation was completely retained in 18 cases, partially removed in 6 cases, and completely removed in 6 cases, while the wound was closed by direct suture in 8 cases, transplanted with gastrocnemius myocutaneous flap in 21 cases, and transplanted with medial sural artery perforator flap in 1 case. Among the patients with postoperative wounds of Achilles tendon rupture, the internal fixation was completely retained in 10 cases and completely removed in 62 cases, and the wound was closed by direct suture in 10 cases and transplanted with sural neurovascular flap in 62 cases. Among the patients with postoperative wounds of calcaneal fracture, the internal fixation was completely removed in 32 cases and completely retained in 13 cases, and the wound was healed by dressing change in 5 cases, closed by direct suture in 5 cases, transplanted with sural neurovascular flap in 23 cases, and transplanted with sural neurovascular flap combined with peroneal brevis muscle flap in 12 cases. Among the patients with postoperative wounds of Pilon fracture, the internal fixation was partially removed in 5 cases, completely retained in 17 cases, and completely removed in 3 cases; the wound was closed by direct suture in 4 cases, transplanted with posterior tibial artery perforator flap in 18 cases, and transplanted with free anterolateral thigh flap in 3 cases. The area of flaps/myocutaneous flaps ranged from 5 cm×3 cm to 18 cm×8 cm, and the area of muscle flaps were from 13.0 cm×1.5 cm to 15.0 cm×2.5 cm. All the wounds closed by direct suture healed. Blood flow obstacle occurred in the distal margin of sural neurovascular flap transplanted in 5 patients and posterior tibial artery perforator flap transplanted in one patient, which healed successfully after dressing change. The other flaps survived well, and the wounds were healed. The patients were followed up for 5 months to 5 years, and no recurrence of infection occurred. The direct suture of the flap donor site left linear scar, and the appearance of the skin graft was good. Three patients with tibial plateau fracture and 2 patients with Pilon fracture had limited joint movement, while the joint activity of the other patients was normal. The patients with Achilles tendon rupture and calcaneal fracture had normal ground motion. The sural neurovascular flap grafted on the wound after calcaneal fracture was bloated resulting in inconvenience in wearing shoes, the gastrocnemius myocutaneous flap grafted on the postoperative wound after tibial plateau fracture was bloated, whereas the appearance of the other flaps was good. Conclusions: For incisional wound infection after orthopedic internal fixation for lower extremity joint injuries, the treatment should be classified according to wound infection and wound site. Dressing change, thorough debridement, reasonable disposal of internal fixation, direct suture after application of VSD, and flap covering, etc. not only ensure the infection control and the wound closure, but also restore the function of the lower limbs to the greatest extent, so as to maximize the benefit of the patient.

[Staged repair strategy for chronic sacrococcygeal radiation ulcer].

Objective: To investigate the clinical effect of staged repair strategy for chronic sacrococcygeal radiation ulcer. Methods: The retrospective cohort study method was applied. Twelve patients with chronic sacrococcygeal radiation ulcer were admitted to Beijing Jishuitan Hospital from January 2010 to June 2020, including 7 males and 5 females, aged 38-74 years. The thorough debridement was performed in the first stage, with wounds area after debridement ranging from 8 cm×6 cm to 22 cm×14 cm, and continuous vacuum sealing drainage (VSD) was performed after the debridement operation. In the second stage, personalized surgery scheme was formulated according to the patient's age, systemic condition, vascular condition, and the position, size, and depth of wound. Six cases were reconstructed with superior/inferior gluteal artery perforator flaps, 4 cases were repaired with gluteus maximus myocutaneous flaps, 1 case was repaired with pedicled latissimus dorsi myocutaneous flap, and 1 case was reconstructed with free transplantation of latissimus dorsi myocutaneous flap. The area of flaps or myocutaneous flaps ranged from 10 cm×8 cm to 25 cm×18 cm. Donor sites of the flaps were sutured primarily in 9 patients and in the other 3 patients were repaired with intermediate split-thickness skin graft in back. The survival of flap or myocutaneous flap after operation, recurrence of tumor, and the appearance and texture of flap or myocutaneous flap, and wound healing were observed during follow-up. Results: Flaps or myocutaneous flaps in 11 patients survived after operation, and superior gluteal artery perforator flap in 1 patient had partial distal necrosis, which was covered again with flap pushed to the distal after debridement and resection of the necrotic tissue. The wounds in 8 patients achieved primary healing, 1 patient repaired with superior gluteal artery perforator flap experienced subcutaneous infection, 1 patient repaired with superior gluteal artery perforator flap suffered distal venous congestion of the flap, and 1 patient repaired with gluteus maximus myocutaneous flap had hematoma under myocutaneous flap, and 1 patient repaired with retrograde latissimus dorsi myocutaneous flap had incision exudation and dehiscence, which were all healed after dressing change, etc. There was no recurrence of tumor after the operation. The wounds healed well during follow-up of 2-52 months after discharge, with no recurrence of infection, and the flaps were soft in texture, with satisfactory appearance and well healed donor sites. Conclusions: On the basis of thorough debridement and VSD in the first stage, superior/inferior gluteal artery perforator flap, gluteus maximus myocutaneous flap, or pedicled/free latissimus dorsi myocutaneous flap with abundant blood supply is applied to repair chronic sacrococcygeal radiation ulcer in the second stage. The staged operation is reliable, with minimal injury to the donor site of flap and satisfactory therapeutic effect.

[Application strategy and clinical effects of paraumbilical perforator flap with inferior epigastric vessels in repairing destructive wounds].

Objective: To explore the application strategy and clinical effects of paraumbilical perforator flap with inferior epigastric vessels in repairing various destructive wounds. Methods: The retrospective observational study method was applied. From January 2015 to December 2020, 28 patients (21 males and 7 females, aged 25 to 66 years) with destructive wounds in various body parts were admitted to Beijing Jishuitan Hospital. The wound areas of patients ranged from 17 cm×8 cm to 35 cm×22 cm after debridement. Pedicled or free paraumbilical perforator flaps with inferior epigastric vessels were used to repair the wounds respectively. The areas of flaps were from 18 cm×10 cm to 37 cm×24 cm, and the lengths of vascular pedicles were 13.0-17.0 (15.1±2.3) cm. For type Ⅲ high-voltage electric burn wounds of wrist, two methods were used to reconstruct the blood flow of hand, one is to bridge the radial artery with saphenous vein grafting and the other one is to design blood flow-through flap. The strength of abdominal wall in the donor site was strengthened by polypropylene patch, and then the wounds were directly sutured. If the wounds could not be sutured directly, then allogenic acellular dermal matrix (ADM) was applied to strengthen the abdominal wall first, and then autologous medium-thickness skin graft was taken from the thigh to cover the wounds. The flap transplantation, hand blood flow reconstruction, the repair of donor site, the flap survival, the wound and donor site healing after operation, the appearance of flaps, and the wound and donor site recovery during follow-up were observed. Results: Among the patients in this group, 13 patients were treated with pedicled flap grafting, while 15 patients were treated with free flap grafting. The hand blood flow of 7 patients with type Ⅲ high-voltage electric burn wounds of wrist was reconstructed by bridging radial artery with saphenous vein grafting. The hand blood flow of 3 patients with type Ⅲ high-voltage electric burn wounds of wrist was reconstructed with blood flow-through flap. In 16 patients, the strength of abdominal wall was strengthened using patch in the donor site,and then the donor sites were sutured directly. In 12 patients, the strength of abdominal wall was strengthened using allogenic ADM, and then the donor sites were covered by skin grafting. All the transplanted flaps survived completely. The wounds of 24 patients were healed, while the wounds of 3 patients with type Ⅲ high-voltage electric burn wounds of wrist and 1 patient with chronic radiation ulcer of ilium failed to heal because of there were still some necrotic tissue and purulent secretion under the flaps. The wounds were healed eventually after debridement and dressing changes. During the follow-up of 6 months to 3 years, the flap survived well with good appearance in all patients, and there was no recurrence, or no abdominal wall hernia occurred in the donor site. Conclusions: Paraumbilical perforator flap with inferior epigastric vessels has flexible design, long vascular pedicle, large area for cut. It can be pedicled or freely transplanted, which is a good choice for repairing destructive wounds in various areas.

Changes in composition a metabolism of caecal microbiota in rats fed diets supplemented with copper-loaded chitosan nanoparticles.

The study was conducted to investigate the effect of copper-loaded chitosan nanoparticles on the composition and metabolism of the caecal microbiota in rats. Forty male Sprague-Dawley (SD) rats (average body weight of 82 ± 5 g) were randomly assigned to five groups (n = 8). The dietary treatments were: (i) basal diet, (ii) basal diet + 80 mg/kg BW CuSO(4), (iii) basal diet + 80 mg/kg BW chitosan (CS-I), (iv) basal diet + 80 mg/kg BW copper-loaded chitosan nanoparticles (CSN-I) and (v) basal diet + 160 mg/kg BW copper-loaded chitosan nanoparticles (CSN-II). The trial lasted 21 days. The results showed that compared with control, Average day gain (ADG) of group CSN-I and CSN-II increased (p < 0.05). No significant difference was found in CuSO(4) or CS-I-treated groups (p > 0.05). There were no effects of these treatments on average day feed intake (ADFI) of rats (p > 0.05). The counts of Bifidobacteria and Lactobacillus in group CSN-II were higher than those of the control group (p < 0.05), while the counts of total aerobes, total anaerobes, Salmonella, Clostridium and coliform were lower than those of the control (p < 0.05). The activity of ß-glucuronidase in group CSN-I or CSN-II was significantly depressed (p < 0.05), while the activities of α-galactosidase and ß-galactosidase were enhanced significantly (p < 0.05). The pH of the caecum digesta and the concentration of propionate and butyrate in group CSN-I and CSN-II were higher than those of the control group (p < 0.05). No significant difference was found in these parameters in CuSO(4) or CS-I-treated groups (p > 0.05). The results indicate that the microbiota and environment of caecum are beneficially changed by the administration of copper-loaded CSN.

[Exploration on the method of aesthetic repair of the donor sites of flaps].

Objective: To explore the excellent methods for aesthetic repair of the donor sites of flaps. Methods: From January 2013 to March 2018, 120 patients (94 males and 26 females, aged from 3 to 60 years) were admitted to the Department of Burns of Beijing Jishuitan Hospital. Wounds areas after debridement or removing scar were ranged from 8.0 cm×3.5 cm to 24.0 cm×18.0 cm. Twenty patients with facial and neck scar were repaired with expanded flaps, including 4 scalp flaps, 8 supraclavicular flaps, 4 deltoid flaps, and 4 trapezius myocutaneous flaps. The flaps in ideal donor sites were selected to repair the wounds in 40 patients, including 20 cases of hand wounds or scars repaired with inguinal flaps, 10 children of foot skin defects or scars repaired with cross inguinal skin flap, 10 cases of knee joint wounds repaired with medial or lateral thigh flaps. The optimal flap design was used to repair wounds in 50 patients. Among the patients, wounds of 36 patients were repaired with relaying flaps, including donor sites of free anterolateral thigh flaps of 8 patients repaired with anteromedial thigh perforator flaps and donor sites of free anterolateral thigh flaps of 8 patients repaired with ilioinguinal flaps or superficial abdominal artery flaps, and donor sites of flaps of 20 patients repaired with peroneal perforator relaying flaps. Besides, wounds of 9 patients were repaired with free lobulated anterolateral thigh flaps, and wounds of 5 patients were repaired with modified V-Y propelling latissimus dorsi myocutaneous flaps. The donor sites of flaps were repaired with allogenic acellular dermal matrix combined with autologous split-thickness skin grafts in 10 cases. The areas of the flaps or myocutaneous flaps were ranged from 6.0 cm×4.0 cm to 30.0 cm×20.0 cm. The survival of flap, myocutaneous flap, or skin graft and the repair of donor site after operation and during follow-up were observed. Results: Blood flow obstacle at 0.5 cm to the distal margin of the flap occurred in 1 patient repaired with expanded flap, which were healed after dressing change. Blood supply disorder occurred at the tip of the anteromedial thigh perforator flap of 1 patient repaired by optimal flap design, which were healed completely after second debridement and restitching. The other flaps or myocutaneous flaps survived well. The allogenic acellular dermal matrix and the autologous split-thickness skin graft survived with good color and texture. During follow-up of 3 months to 4 years, the donor sites of flaps had good appearance, only with linear scar and the function recovered well. The donor sites of skin grafts had no scar hyperplasia, only with scattered pigmentation. Conclusions: According to the characteristics of donor sites of flaps, individualized and reasonable design before the operation such as pre-expanding of the flaps, selecting the ideal donor sites, optimization of the flap design or allogenic acellular dermal matrix combined with autologous split-thickness skin graft to repair donor sites of flaps can minimize the damage for function and appearance of donor sites of flaps and achieve aesthetic effects of donor sites of flaps.

[Epidemiological investigation of 169 inpatients with high-voltage electrical burns on the wrist].

Objective: To analyze the epidemiological characteristics of inpatients with high-voltage electrical burns on the wrist in Beijing Jishuitan Hospital (hereinafter referred to as the author's unit), so as to provide reference for the prevention and treatment of high-voltage electrical burns on the wrist. Methods: The medical records of inpatients suffered from high-voltage electrical burns on the wrist in the author's unit from January 2008 to December 2019 were collected. The patients' gender, age, population category, injury situation, injury season, total burn area, electrical burn type on the wrist of the affected limbs, the time from injury to first flap/myocutaneous flap transplantation, vascular reconstruction and flap/myocutaneous flap transplantation of the affected limbs, the total amputation rate of the affected limbs and the amputation rate of the affected limbs with type Ⅲelectrical burns on the wrist, the number of operation, postoperative infection rate of flap/myocutaneous flap, length of hospital stay, hospitalization expense, and treatment outcome were retrospectively analyzed. Comparison of the aforementioned statistical items between patients admitted from January 2008 to December 2013 (hereinafter referred to as the pre-stage) and January 2014 to December 2019 (hereinafter referred to as the post-stage) except gender, the total amputation rate of the affected limbs, treatment outcome were performed. Data were statistically analyzed with chi-square test, Fisher's exact probability test, and Wilcoxon rank-sum test. Results: During the 12 years, a total of 169 patients with high-voltage electrical burns on the wrist were admitted to the author's unit, including 162 males and 7 females, aged (35±13) years, and 75.15% (127/169) of patients were 21-50 years old. The top three groups in population category from high to low were workers, migrant workers, and primary and secondary school students, accounting for 48.52% (82/169), 28.99% (49/169), and 9.47% (16/169), respectively. At work of non-electric power accounted for 47.93% (81/169) and ranked the first in the proportion of injury situation. The injury occurred mostly in summer and autumn, accounting for 39.05% (66/169) and 28.99% (49/169), respectively. About 65.09% (110/169) of the patients were with total burn area less than 10% total body surface area. There were totally 216 affected limbs with high-voltage electrical burns on the wrist, of which the numbers of wrist with type Ⅲ and type Ⅳ injury were 25 (11.57%) and 21 (9.72%), respectively. The time from injury to first flap/myocutaneous flap transplantation was 6.00 (3.75, 8.00) d. There were 45 affected limbs operated with vascular reconstruction, 75 affected limbs transplanted with pedicled axial flap, and 86 affected limbs transplanted with free flap/myocutaneous flap. The total amputation rate of affected limbs was 12.96% (28/216), and the amputation rate of the affected limbs with type Ⅲ electrical burns on the wrist was 28% (7/25). The number of operation was 4 (3, 5) times, the postoperative infection rate of flap/myocutaneous flap was 7.18% (13/181), the hospitalization time was 39.00 (25.00, 50.00) d, and the hospitalization cost was 123 553.00 (50 656.50, 216 003.00) yuan. Compared with those of the pre-stage, the time from injury to first flap/myocutaneous flap transplantation was significantly shortened (Z=-4.038, P<0.01), the ratio of free flap/myocutaneous flap transplantation on affected limbs was significantly increased (χ(2)=13.478, P<0.01), the ratio of pedicled axial flap transplantation on affected limbs was significantly decreased (χ(2)=10.242, P<0.01), the number of operation was significantly reduced (Z=-5.903, P<0.01), the postoperative infection rate of flap/myocutaneous flap was significantly decreased (χ(2)=4.492, P<0.05), the length of hospital stay was significantly shortened (Z=-2.723, P<0.01), and the hospitalization expense was significantly decreased among patients hospitalized in the post-stage (Z=-2.121, P<0.05). The other items were close between patients hospitalized in the pre-stage and those in the post-stage. Conclusions: Men were more likely than women to suffer from high-voltage electrical burns on the wrist in the author's unit. Young workers and migrant workers may be the key groups for the prevention of high-voltage electrical burns on the wrist. It is very important to strengthen the management of high voltage safety education for non-electric power staff, warn and protect the surrounding area of high voltage environment, and prohibit non staff for entering the high voltage environment, especially in summer and autumn. In the last 6 years, the patients with high-voltage electrical burns on the wrist may benefit from the active vascular reconstruction at early stage and free flap/myocutaneous flap repair in the author's unit.

[Limb salvage strategies for patients with high voltage electric burns of extremities on the verge of amputation].

Objective: To explore the limb salvage strategies for patients with high voltage electric burns of extremities on the verge of amputation. Methods: From January 2003 to March 2019, 61 patients with high voltage electric burns of extremities on the verge of amputation were treated in our hospital. All of them were male, aged 15-58 years, including 49 cases of upper limbs and 12 cases of lower limbs. The wound area after thorough debridement ranged from 15 cm×11 cm to 35 cm×20 cm. Emergency surgery for reconstruction of the radial artery with saphenous vein graft under eschar was performed in 5 cases. The arteries of 36 patients (including 7 cases with simultaneous ulnar artery and radial artery reconstruction) were reconstructed with various forms of blood flow-through after debridement, among them, the radial artery of 13 cases, the ulnar artery of 8 cases, the brachial artery of 8 cases, and the femoral artery of 2 cases were reconstructed with saphenous vein graft; the radial artery of 3 cases and the ulnar artery of 7 cases were reconstructed with the descending branch of the lateral circumflex femoral artery graft; the radial artery of 2 cases were reconstructed with greater omentum vascular graft; the reflux vein of 3 cases with wrist and forearm annular electric burns were reconstructed with saphenous vein graft. According to the actual situation of the patients, 12 cases of latissimus dorsi myocutaneous flap, 6 cases of paraumbilical flap, 28 cases of anterolateral thigh flap, 10 cases of abdominal combined axial flap, 5 cases of greater omentum combined with flap and/or skin grafts were used to repair the wounds after debridement and cover the main wounds as much as possible. Some cases were filled with muscle flap in deep defect at the same time. The area of tissue flaps ranged from 10 cm×10 cm to 38 cm×22 cm. For particularly large wounds and annular wounds, the latissimus dorsi myocutaneous flap, the paraumbilical flap, the abdominal combined axial flap, and the greater omentum combined with flap and/or skin grafts were used more often. Donor sites of three patients were closed directly, and those of 58 patients were repaired with thin and medium split-thickness skin or mesh skin grafts. The outcome of limb salvage, flap survival, and follow-up of patients in this group were recorded. Results: All the transplanted tissue flaps survived in 61 patients. Fifty-six patients had successful limb salvage, among them, 31 limbs were healed after primary surgery; 20 limbs with flap infection and tissue necrosis survived after debridement and flap sutured in situ; 5 limbs with flap infection, radial artery thrombosis, and hand blood supply crisis survived after debridement and radial artery reconstruction with saphenous vein graft. Five patients had limb salvage failure, among them, 3 patients with wrist electric burns had embolism on the distal end of the transplanted blood vessels, without condition of re-anastomosis, and the hands gradually necrotized; although the upper limb of one patient was salvaged at first, due to the extensive necrosis and infection at the distal radius and ulna and the existence of hand blood supply under flap, considering prognostic function and economic benefits, amputation was required by the patient; although the foot of one patient was salvaged at first, due to the repeated infection, sinus formation, extensive bone necrosis of foot under flap, dullness of sole and dysfunction in walking for a long time, amputation was required by the patient. During the follow-up of 6 months to 5 years, 56 patients had adequate blood supply in the salvaged limbs, satisfied appearance of flaps, and certain recovery of limb function. Conclusions: Timely revascularization, early thorough debridement, and transplantation of large free tissue flap, combined tissue flap, or blood flow-through flap with rich blood supply are the basic factors to get better limb preservation and recovery of certain functions for patients with high voltage electric burns of limbs on the verge of amputation.

[Reconstruction of perineal obliteration deformity after extensive deep burn with ilioinguinal flap].

Objective: To explore the effect of ilioinguinal flap on reconstruction of perineal obliteration deformity after extensive deep burn. Methods: Five patients with perineal obliteration deformity after extensive deep burn were hospitalized from January 2010 to June 2015, with total burn area ranging from 35% to 55% total body surface area, depth of full-thickness burn and wound deep to bone, and course of scar from 6 months to 3 years. Scars of patients were involved in bilateral groins, inner thighs, monsveneris, sacrococcygeal region, and central area of perineum. The abduction angles of double lower limbs ranged from 30 to 65°. Anus was narrow, and defecation was difficult. After release of scar tissue in perineal region, the wound area ranged from 23 cm×12 cm to 28 cm×24 cm. For wound repair and reconstruction of anus, unilateral ilioinguinal flap was used in 3 cases. Due to large wound in two patients, bilateral ilioinguinal flap was used in one patient, and unilateral ilioinguinal flap combined with anterolateral femoral flap was used in another one patient. The area of unilateral ilioinguinal flap ranged from 23 cm×12 cm to 30 cm×20 cm, and the area of anterolateral femoral flap was 21 cm×12 cm. The abdominal donor site was closed with partial suture and partial skin grafting (harvested from split-thickness skin of autologous head or thin intermediate-thickness skin of autologous back). The femoral donor site was directly sutured. After the operation, the double lower limbs were fixed with plaster on abducent position and strictly immobilized. Results: All the flaps survived after operation and the wounds healed well. During the follow-up for 6 to 12 months, the appearance of flaps were good with soft texture and no contracture. Hip joint motion was good, and abduction angles of double lower limbs ranged from 110 to 135°. The appearance of crissum was good without skin inflammation and with normal function of defecation. The appearance of donor site was acceptable to patients or their parents. Conclusions: Ilioinguinal flap is a good choice for reconstruction of perineal obliteration deformity after burn.

Effects of cadmium on serum sex hormone levels in pigs.

An inhibition of sex hormone secretion is observed in domestic animals exposed to cadmium in experimental conditions. Ninety-six gilts (Duroc x Landrace x Yorkshire, initial weight 27.59 kg) were randomly assigned into four different groups with three replications in each group (eight gilts per replication). The groups received the corn-soybean basal diet and supplemented with 0, 0.5, 5.0 and 10.0 mg/kg cadmium (as CdCl2) respectively. The feeding trial was ended when body weight of pigs was approximately 90 kg. The results showed that 10.0 mg/kg cadmium significantly decreased average daily gain (p < 0.05) and increased feed/gain ratio (p < 0.05) of pigs. Follicle-stimulating hormone, luteinizing hormone, oestradiol (E2) and progesterone levels in serum of the pigs fed the diet supplemented with 10.0 mg/kg cadmium were lower significantly than those of the control (p < 0.05). Compared with the control, no changes were found in the levels mentioned above when the pigs fed the diet supplemented with 0.5 or 5.0 mg/kg cadmium (p > 0.05). The changes of regulatory mechanism of the pituitary-ovary axis may be the result of cadmium accumulation in pituitary and ovary and should be investigated further.

Characterization of aflJ, a gene required for conversion of pathway intermediates to aflatoxin.

The genes encoding the aflatoxin biosynthetic pathway enzymes have been localized as a cluster to a 75-kb DNA fragment. The enzymatic functions of the products of most of the genes in the cluster are known, but there are a few genes that have not yet been characterized. We report here the characterization of one of these genes, a gene designated aflJ. This gene resides in the cluster adjacent to the pathway regulatory gene, aflR, and the two genes are divergently transcribed. Disruption of aflJ in Aspergillus flavus results in a failure to produce aflatoxins and a failure to convert exogenously added pathway intermediates norsolorinic acid, sterigmatocystin, and O-methylsterigmatocystin to aflatoxin. The disrupted strain does, however, accumulate pksA, nor-1, ver-1, and omtA transcripts under conditions conducive to aflatoxin biosynthesis. Therefore, disruption of aflJ does not affect transcription of these genes, and aflJ does not appear to have a regulatory function similar to that of aflR. Sequence analysis of aflJ and its putative peptide, AflJ, did not reveal any enzymatic domains or significant similarities to proteins of known function. The putative peptide does contain three regions predicted to be membrane-spanning domains and a microbodies C-terminal targeting signal.