Objective:This study aimed to investigate the role and clinical significance of MUC4 gene mutations in thrombotic events in patients with classic paroxysmal nocturnal hemoglobinuria (PNH) patients.Methods:A retrospective analysis was conducted on the clinical data and gene sequencing results of 45 patients with classic PNH admitted to the Department of Hematology, Tianjin Medical University General Hospital, from June 2018 to February 2022. MUC4 gene mutations in patients with classic PNH were summarized, and the risk factors for thrombotic events in these patients were analyzed. Additionally, the effects of MUC4 gene mutations on the cumulative incidence and survival of thrombotic events in patients with classic PNH were determined.Results:The detection rate of MUC4 gene mutations in patients with classic PNH who experienced thrombotic events (thrombotic group) was 68.8% (11/16), which was significantly higher than that in the non-thrombotic group [10.3% (3/29) ] ( P<0.001). All mutations occurred in exon 2. MUC4 mutation ( OR=20.815, P=0.010) was identified as an independent risk factor for thrombotic events in patients with classic PNH. The cumulative incidence of thrombotic events was 78.6% (11/14) in the MUC4 gene mutation group (mutation group) and 16.1% (5/31) in the non-mutation group, showing a statistically significant difference between the two groups ( P<0.001). Survival analysis showed a lower overall survival (OS) rate in the thrombotic group compared with that in the non-thrombotic group [ (34.4±25.2) % vs. (62.7±19.3) % ] ( P=0.045). The OS rate of patients was (41.7±29.9) % in the mutation group and (59.1±18.3) % in the non-mutation group ( P=0.487) . Conclusion:MUC4 gene mutations are associated with an increased incidence of thrombotic events in classic PNH patients, highlighting their role as independent risk factors for thrombosis in this population. These mutations can be considered a novel predictive factor that aids in evaluating the risk of thrombosis in patients with classic PNH.

Since microdroplets are able to be generated rapidly in large amount and each droplet can be well controlled as an independent micro-cultivator, droplet microfluidic technology can be potentially used in the culture of microorganisms, and provide the microbial culture with high throughput manner. But its application mostly stays in the laboratory-level building and using for scientific research, and the wide use of droplet microfluidics in microbial technology has been limited by the key problems that the operation for microdroplets needs high technical requirements with wide affecting factors and the difficulties in integration of automatic microdroplet instrumentation. In this study, by realizing and integrating the complicated operations of droplet generation, cultivation, detection, splitting, fusion and sorting, we design a miniaturized, fully automated and high-throughput microbial microdroplet culture system (MMC). The MMC can be widely used in microbial growth curve test, laboratory adaptive evolution, single factor and multi-level analysis of microbial culture, metabolite detection and so on, and provide a powerful instrument platform for customized microbial evolution and screening aiming at efficient strain engineering.
Industrial Development ; Microfluidics

Objective:To explore the application value of skin lead marker combined with iSCOUT image-guided positioning system in monitoring and correcting the setup error of intensity-modulated radiotherapy (IMRT) for breast cancer and calculate the PTV margin, aiming to provide reference for clinical practice.Methods:25 breast cancer patients treated with IMRT after modified radical mastectomy in Fujian Medical University Union Hospital from April to August 2019 were enrolled in this study. The skin lead marker combined with iSCOUT image-guided positioning system was employed for image-guided positioning based on the gold standard registration algorithm. Initial setup errors on the x (lateral), y (craniocaudal) and z (anteroposterior) axis and residual errors after the position correction were recorded and analyzed. The effect of the errors before and after image-guided correction upon the plan dose was compared and the reasonable PTV margin was calculated.Results:25 patients received 150 times of positioning verification using skin lead marker combined with iSCOUT image-guided positioning system. The absolute residual errors on the x-, y-and z-axis were (1.53±0.96), (1.30±0.99) and (1.34±0.92) mm, significantly smaller than the initial setup errors of (2.63±2.12), (2.41±2.45) and (3.07±2.77) mm (all P<0.001). The percentage of dose deviation due to residual errors was also smaller than that of the initial errors. Significant differences were observed in D 98%, D 2%, D max of PTV, D max of the heart, D max of the healthy breast, and D mean of the affected lung and both lungs. The percentage deviation from the original plan was decreased from 2.18%, 3.19%, 10.66%, 8.75%, 48.21%, 10.50%, and 3.66% to 0.38%, 0.23%, 2.31%, 0.04%, 13.78%, 6.35% and 0.41%, respectively (all P<0.05). PTV margins on the x-, y-and z-axis after correction were calculated as 1.87, 1.75 and 1.69 mm, respectively. Conclusion:It is feasible and valuable to apply the skin lead marker combined with iSCOUT image-guided positioning system in the positioning verification and correction of breast cancer radiotherapy position, providing novel reference for clinical PTV margin.

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at 28°C, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ΔAcuD was significantly attenuated in this Zebrafish infection model.
Gene Knockout Techniques ; Hyphae ; Isocitrate Lyase ; Microinjections ; Mortality ; Rhombencephalon ; RNA Interference ; Talaromyces ; Virulence ; Yeasts ; Zebrafish

Objective To explore the perioperative outcomes and safety of enhanced recovery after surgery (ERAS) in laparoscopic radical cystectomy (LRC).Methods We retrospectively evaluated outcome of 10 LRC patients on ERAS protocol from May 2017 to October 2017,and 39 LRC patients on conventional recovery after surgery(CRAS) protocol from July 2015 to November 2016.There were (60.9 ±11.4) years and (63.7 ± 12.1) years in ERAS group and CRAS group respectively(P =0.514);(25.5 ±2.7) kg/m2 and (24.4 ± 3.6) kg/m2 with body mass index (P =0.375).Both of the median of charlson comorbidity index (P =0.931) and American Society of Anesthesiologists score (P =0.254) were 2 There was no statistical significance between the two groups for type of urinary diversion and preoperative laboratory studies (P ＞ 0.05).Patients' perioperative outcomes,early (30-day) complications and postoperative readmission rate were compared.Results The ERAS group had less intraoperative crystalloid infusion [(950.0 ± 474.3) ml vs.(1 797.4 ± 448.1) ml,P ＜ 0.001],faster removed gastric tube (0 d vs.4 d,P ＜0.001),and shorter passing flatus time [(1.6 ± 0.8) d vs.(2.9 ± 1.4) d,P =0.006] than the CRAS group;however,no difference was found in terms of intraoperative colliod infusion [(1 110.0 ± 331.5)ml vs.(1 117.9 ± 397.9) ml,P =0.954].No patients from either group required conversion to open surgery.There was no significant difference between the two groups for operative time (P =0.311),estimated blood loss (P =0.073),drain days (P =0.681),postoperative hospital stay (P =0.509),overall blood transfusion (P =1.000),intensive care unit stay (P =1.000) and tumor characteristics (pathological stage,histology,nodes removed,positive nodes,lymph node-positive patients,positive surgical margins).The 30-day postoperative complications were documented in 5 (50％)and 23 (59％)patients in groups ERAS and CRAS (P =0.878),respectively.And the most common complication were minor complications (Clavien-Dindo grade 1 and 2) in both groups (100.0％ vs.86.9％,P =0.729).The 30-day readmission rate was 20.0％ (2 patients) in ERAS group and 10.3％ (4 patients) in CRAS group with no statistical significance(P =0.588).Conclusions Our ERAS protocol expedited bowel function recovery after RC and urinary diversion without increasing in 30-day complications compared with CRAS.The key of implement ERAS pathway is to explore and develop their own protocol conformed to their medical treatment enviroment.

Objective To investigate the effect of metformin combined with acupoint therapy with low frequency impulse stimulation on repairing diabetic neuropathy of patients with diabetic foot.Methods Totally 150 diabetic foot patients with neuropathy were randomly divided into observation group (n =75) and control group (n =75).The therapeutic effect,nerve conduction velocity,fasting blood glucose,postprandial 2 h blood glucose,glycosylated hemoglobin and adverse reactions were compared between the two groups.Results The total effective rate of the observation group was 92％,which was significantly higher than 76％ of the control group (P ＜0.05).The SCV and MCV of the tibial nerve and peroneal nerve increased significantly after treatment in all the patients (P ＜0.05),and the SCV and MCV of the tibial nerve and peroneal nerve in the observation group were significantly higher than those in the control group (P ＜ 0.05).After treatment,the levels of HbA1 C,FBG and 2 hPBG decreased significantly when compared with those before treatment (P ＜ 0.05),and the levels of HbA1 C,FBG and 2 hPBG in the observation group were significantly lower than those in the control group (P ＜ 0.05).The total incidence rate of complications in the observation group was 8％,which was significantly lower than 20％ in the control group (P ＜ 0.05).Conclusion Metformin combined with acupoint therapy with low frequency impulse stimulation can effectively improve the symptoms of neuropathy in patients with diabetic foot.

Objective To investigate the effect of metformin combined with acupoint therapy with low frequency impulse stimulation on repairing diabetic neuropathy of patients with diabetic foot.Methods Totally 150 diabetic foot patients with neuropathy were randomly divided into observation group (n =75) and control group (n =75).The therapeutic effect,nerve conduction velocity,fasting blood glucose,postprandial 2 h blood glucose,glycosylated hemoglobin and adverse reactions were compared between the two groups.Results The total effective rate of the observation group was 92％,which was significantly higher than 76％ of the control group (P ＜0.05).The SCV and MCV of the tibial nerve and peroneal nerve increased significantly after treatment in all the patients (P ＜0.05),and the SCV and MCV of the tibial nerve and peroneal nerve in the observation group were significantly higher than those in the control group (P ＜ 0.05).After treatment,the levels of HbA1 C,FBG and 2 hPBG decreased significantly when compared with those before treatment (P ＜ 0.05),and the levels of HbA1 C,FBG and 2 hPBG in the observation group were significantly lower than those in the control group (P ＜ 0.05).The total incidence rate of complications in the observation group was 8％,which was significantly lower than 20％ in the control group (P ＜ 0.05).Conclusion Metformin combined with acupoint therapy with low frequency impulse stimulation can effectively improve the symptoms of neuropathy in patients with diabetic foot.

Objective: To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Methods: Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA⁺ nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA⁺ nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA⁺ nucleated RBC was detected by Western blot. Results: Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA⁺ nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA⁺ nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA⁺ nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA⁺ nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA⁺ nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Conclusion Autophagy is impaired in nucleated RBC of MDS patients.

Objective To assess the clinical effectiveness and safety of relinqing pharmaceutical preparations for the treatment of uncomplicated urinary tract infection(UTI). Methods The genitourinary infection, urinary tract infection, pyelonephritis, cystitis, stranguria and urethritis were used as key words to search at CNKI,VIP,SinoMed,PubMed,Wan Fang and Cochrane Library Databases up to April 2015. Data of randomised controlled trials (RCTs) comparing treatments using relinqing were included in this study. The quality of the literature was evaluated by the method of Cochrane handbook 5.1.0. Data extraction was carried out independently by two authors. RevMan 5.2 software was used for Meta-analysis. Results Five RCTs were included that involved a total of 471 uncomplicated UTIs. Analysis of four studies showed a higher rates of effectiveness for uncomplicated UTI in the treatment with relinqing plus antibiotics than those of antibiotics alone [RR and 95%CI:1.15 (1.08-1.23), P<0.001]. Analysis of two studies showed a higher rates of bacterial clearance for uncomplicated UTI in the treatment with relinqing plus antibiotics than those of antibiotics alone [RR and 95% CI: 4.04 (1.78-9.16)]. Conclusion Data from five small studies suggest that relinqing as an independent intervention or in conjunction with antibiotics may be beneficial for treating uncomplicated UTIs. However, the small number and poor quality of the included studies meant that it is not possible to formulate robust conclusion on the use of relinqing for uncomplicated UTI either alone or as an adjunct to antibiotics.

OBJECTIVETo explore the expression levels of terminal complement complex (C5b-9) and CD62p on platelets and the soluble C5b-9 (sC5b-9) level in serum in patients with PNH or PNH-aplastic anemia (AA).

METHODSSerum levels of sC5b-9, complement C3 and C4 were detected by using ELISA in 25 patients with PNH/PNH-AA. The quantities of C5b-9 and CD62p on the membrane of platelets were detected by flow cytometry.

RESULTS①In PNH/PNH-AA group, the serum sC5b-9 level [390.27(265.73-676.87) μg/L] was lower than that in control group [540.39(344.20-1 576.78) μg/L] (P<0.01). ②The platelet PNH clone (CD59⁻CD61⁺/CD61⁺) size [50.58(23.29-81.60)%] was bigger in the PNH/PNH-AA group than that [23.57(15.58-29.02)%] in control group (P<0.01). The percentages of C5b-9 deposition (C5b-9⁺CD61⁺/CD61⁺) were higher on the PNH clone platelets (CD59⁻CD61⁺) in the PNH/PNH-AA group [(17.53 ± 6.27)%] than those on the normal platelets (CD59⁺CD61⁺) in PNH patients 11.33±5.03）%］ and control [(10.88±3.58)%] group (P<0.01). ③ The expression of CD62p (CD62p⁺CD61⁺/CD61⁺) on PNH clone platelets in PNH patients [(61.98 ± 11.71)%] was higher than that on the normal platelets in PNH patients [(43.76±11.30)%] and control group [(38.23±18.07)%] (P<0.01). In addition, the expression of CD62p on normal platelets was higher in PNH patients than control (P<0.05). ④The deposition of C5b-9 positively correlated with the expression of CD62p on the platelets (r=0.559, P=0.002).

CONCLUSIONDeficiency of CD59 antigen on platelets in PNH patients may lead to the deposition of C5b-9 on its membrane and its dysfunction, which may contribute to thrombosis events in PNH.