Objective: To identify the phenotypes, antibodies and explore the molecular mechanisms of a patient who carries antibodies to RBC high-frequency antigens and his family members. Methods: The antibody identification test was performed for the proband by serological methods, and targeted NGS was subsequently used to detect mutations that occurred in blood group genes. Blood samples were collected from the proband and his family members. Sanger sequencing was used to verify the mutation of the XK gene. The expression of Kell blood group antigens was detected by serological methods and flow cytometry. K cells were used to detect the antibody specificity of the proband. The morphology of red blood cells was detected by the scanning electron microscopy. The serum creatine kinase levels of the proband and his family members were analyzed by colorimetric methods. Results: The results of the antibody identification test suggested that the proband might have antibodies to high-frequency antigens. NGS results suggested a homozygous mutation (c. 107G>A) in exon 1 of the XK gene in the proband, resulting in a truncated XK protein. The Sanger sequencing results of the proband were consistent with the NGS results, and the mutation was not found in other family members. The expression of Kell blood group antigens of the proband was not found by serological methods and flow cytometry. The results of the antibody specificity test showed that the proband had anti-Km antibodies. Spike-like changes were identified on red blood cells, and serum creatine kinase level was elevated in the proband. Conclusion: In this study, the McLeod phenotype caused by homozygous mutation (c. 107G>A) of XK gene was identified in Chinese individuals for the first time by the phenotype and molecular mechanism studies. The results of genotyping and phenotyping suggested that the McLeod phenotype caused by the mutation was compatible with the phenotypes of McLeod and K .

Objective:To explore the assessment value of liver enzyme changes before and after hepatectomy on the prognosis of patients with primary hepatocellular carcinoma(HCC). Methods:A retrospective analysis was conducted on the clinical data of 421 patients with primary HCC who received at the First and Second Affiliated Hospitals of Xi'an Jiaotong University from January 2016 to December 2020.The univariate and multivariate COX regression analyses were used to screen the risk factors affecting the prognosis of patients with primary HCC,and the optimal cut-off value was selected to group the most valuable indicators among them,further analyzing the different factors among subgroups.The Kaplan-Meier method was used to assess the prognosis of the subgroups,and the survival curves were plotted and statistically tested. Results:Multivariate COX regression analysis revealed that Barcelona Clinic Liver Cancer(BCLC)stage,tumor number,alpha-fetoprotein(AFP),the quantitative change of γ-glutamyl transpeptidase(GGT)and total bilirubin(TBIL)before and after surgery were independent risk factors affecting the prognosis of patients with primary HCC(P＜0.01),and there were differences in hepatitis B virus surface antigen(HBsAg),TNM stage,BCLC stage,tumor size,tumor number,tumor differentiation degree,AFP,surgery type,intraoperative bleeding volume,postoperative albumin-bilirubin(ALBI)and white blood cell count between the high and low GGT variation groups(P＜0.05).Kaplan-Meier survival analysis showed that the prognosis of patients with primary HCC between the high and low GGT variation groups was significantly different,with a hazard ratio of 2.603 for mortality and 1.449 for recurrence(both P＜0.01). Conclusion:The quantitative change of GGT before and after hepatectomy has an evaluation value for the prognosis of patients with primary HCC,and the quantitative change of GGT greater than 29.5 is an independent risk factor for the prognosis of patients with primary HCC.

OBJECTIVE: To report the clinical and genetic characteristics of a rare case of Gitelman syndrome with comorbid Graves disease and ACTH-independent adrenocortical adenoma. METHODS: A patient who had presented at the Nanchong Central Hospital on December 21, 2020 was selected as the study subject. Clinical data of the patient was collected. Whole-exome sequencing was carried out on DNA extracted from peripheral venous blood samples from the patient and her family members. RESULTS: The patient, a 45-year-old woman, was found to have Graves disease, ACTH-independent Cushing syndrome, hypokalemia and hypomagnesemia following the discovery of an adrenal incidentaloma. MRI scan had revealed a 3.8 cm × 3.2 cm mass in the left adrenal gland. The mass was removed by surgery and confirmed as adrenocortical adenoma. DNA sequencing revealed that the patient and her sister have both harbored compound heterozygous variants of the SLC12A3 gene, namely c.1444-10(IVS11)G>A and c.179(exon1)C>T (p.T60M), which were respectively inherited from their father and mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1444-10(IVS11)G>A and c.179(exon1)C>T (p.T60M) were respectively classified as a variant of uncertain significance (PM2_Supporting+PP3) and a likely pathogenic variant (PM3_Strong+PM1+PP3). CONCLUSION The conjunction of Gitelman syndrome with Graves disease and adrenal cortex adenoma is rather rare. The newly discovered c.1444-10(IVS11)G>A variant of the SLC12A3 gene, together with the heterozygous variant of c.179(exon1)C>T (p.T60M), probably underlay the pathogenesis in this patient.
Humans ; Female ; Middle Aged ; Gitelman Syndrome/genetics* ; Adrenocortical Adenoma ; Hypokalemia ; Graves Disease/genetics* ; Mothers ; Mutation ; Solute Carrier Family 12, Member 3

Objective:To assess the efficacy and safety of 100 units of botulinum toxin A (BTX-A) intradetrusor injection in patients with overactive bladder.Methods:From April 2016 to December 2018, 17 tertiary hospitals were selected to participate in this prospective, multicenter, randomized, double-blind, placebo-controlled study. Two phases of study were conducted: the primary phase and the extended phase. This study enrolled patients aged 18 to 75 years who had been inadequately managed by anticholinergic therapy (insufficient efficacy or intolerable side effects) and had spontaneous voiding with overactive bladder. Exclusion criteria included patients with severe cardiac, renal and hepatic disorders, patients with previous botulinum toxin treatment for 6 months or allergic to BTX-A, patients with urinary tract infections, patients with urinary stones, urinary tract tumors, diabetes mellitus, and bleeding tendency. Eligible patients were randomly assigned to BTX-A group and placebo control group in a ratio of 2∶1. Two groups of patients received 20 intradetrusor injections of BTX-A 100U or placebo at the depth of the submucosal muscle layer respectively under cystoscope, including 5 injections at the base of the bladder, 3 injections to the bladder triangle, 5 injections each to the left and right walls and 2 injections to the top, sparing the bladder neck. As a placebo control group, patients received same volume of placebo containing no BTX-A and only adjuvant freeze-dried preparations for injection with the same method. A combination of gelatin, sucrose, and dextran served as adjuvants. Average micturition times per 24 hours, urinary incontinence (UI) episodes per day, average micturition volume per day, OAB symptom score(OABSS), and quality of life (QOL) score were recorded at baseline and the 2nd, 6th and 12th week after treatment. The primary efficacy endpoint was the change from baseline in the average micturition times per 24 hours at the 6th week after treatment. The secondary efficacy endpoints included the change from baseline in the average micturition times per 24 hours at 2nd and 12th week, as well as the change from baseline in the OABSS, QOL score, average frequency of urgency and UI episodes per day, urgency score, average micturition volume per day at 2nd, 6th and 12th week after treatment. Patients were followed for 12 weeks to assess adverse events (AEs). After assessed at week 12, if the micturition times has decreased less than 50% compared to baseline and the patient is willing to receive retreatment, then patients could enter the extended trial phase. In that phase, patients in both groups were injected with 100 units BTX-A from 12th week onwards and then followed up the same indicators for 12 weeks.Results:216 patients were enrolled in this trial (144 cases in the BTX-A group and 72 cases in the placebo control group). Baseline characteristics such as age (47.75±14.20 in the BTX-A group and 46.39±15.55 in the control group), sex (25 male/117 female in the BTX-A group and 10/61 in the control group), and disease duration (0.51 years in the BTX-A group and 0.60 years in the control group) were balanced between the two groups( P>0.05). A marked reduction from baseline in average micturition times per 24 hours was observed in all treatment groups at the 6th week and the reduction of the two groups was statistically different ( P<0.001 and P=0.008 respectively). Compared with the baseline, the average micturition times per 24 hours at the 6th week decreased from baseline by 2.40(0.70, 4.60)times for the BTX-A group and 0.70(-1.00, 3.30) times for the placebo control group respectively, and the difference between the two groups was considered to be statistically significant ( P=0.003). The change rates of average micturition times per 24 hours from baseline at the 6th week of the two groups were (16±22)% and (8±25)% respectively, and the difference between the two groups was statistically significant ( P=0.014). Compared with the baseline, the average micturition times per 24 hours at 2nd and 12th week decreased by 2.00(0.00, 4.00)and 3.30(0.60, 5.03)for the BTX-A group, 1.00(-1.00, 3.00)and 1.70(-1.45, 3.85)for the placebo control group respectively. The difference between two groups was considered to be statistically significant ( P=0.038 and P=0.012); the changes of average urgency times per day for the BTX-A group and the control group at the 2nd, 6th and 12th week were 2.00(0.00, 4.30)and 2.40(0.30, 5.00), 3.00(0.30, 5.70)and 0.70(-1.30, 2.70), 0.70(-1.30, 3.00) and 1.35(-1.15, 3.50), respectively. There were significant differences between two groups at the 2nd, 6th and 12th week, ( P=0.010, P=0.003 and P=0.025, respectively). The OABSS of the BTX-A group and the control group at the 6th week decreased by 1.00(0.00, 4.00)and 0.50(-1.00, 2.00) compared with the baseline, and the difference between the two groups was statistically significant ( P=0.003). 47 cases of BTX-A group and 34 cases of placebo control group entered the extended trial phase, and 40 and 28 cases completed the extended trial phase, respectively. The average micturition volume per 24 hours changed by -16.60(-41.60, -0.60)ml and -6.40(-22.40, 13.30)ml, (-35.67±54.41)ml and(-1.76±48.69)ml, (-36.14±41.51)ml and (-9.28±44.59)ml, (-35.85±43.35)ml and(-10.41±40.29)ml for two groups at the 12th, 14th, 18th and 24th week, and the difference between two groups was statistically significant at each follow-up time ( P=0.01, 0.006, 0.012 and 0.016, respectively). There was no significant difference in other parameters( P>0.05). However, adverse reactions after intradetrusor injection included increased residual urine volume (27 in the BTX-A group and 3 in the control group), dysuria (21 in the BTX-A group and 6 in the control group), urinary infection (19 in the BTX-A group and 6 in the control group), bladder neck obstruction (3 in the BTX-A group and 0 in the control group), hematuria (3 in the BTX-A group and 1 in the control group), elevated alanine aminotransferase (3 in the BTX-A group and 0 in the control group), etc. During the follow-up period, there was no significant difference in the other adverse events between two groups except the increase of residual urine volume( P<0.05). In the primary trial phase, among the 27 cases with increased residual urine volume in BTA group, only 1 case (3.70%) with PVR more than 300 ml; the PVR of 3 patients in the placebo group was less than 100 ml. The increase of residual urine volume caused by the injection could be improved or disappeared with the passage of time. Conclusions:Intradetrusor injection of Chinese BTX-A improved the average micturition times per 24 hours, the average daily urgent micturition times, OABSS, and average micturition volume per time, and reduced the adverse effects in patients with overactive bladder.Chinese BTX-A at dose of 100U demonstrated durable efficacy and safety in the management of overactive bladder.

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

Objective To investigate the effect of sodium houttuyfonate on the expression of PI3K and AKT1 and mTOR mRNA in the lung of rats with chronic obstructive pulmonary disease(COPD),and reveal the possible mechanism of the COPD treated with sodium houttuyfonate. Methods Twenty-four male Wistar rats were randomly divided into normal control group,model control group,dexamethasone group and sodium houttuyfonate group(n=6 for each). The rat models of COPD were established by intratracheal instillation of lipopolysaccharide and smudging. The expressions of PI3K and AKT1 and mTOR mRNA were determined by real-time PCR. The morphological changes of the lung tissue was examined by histopathology. Results Compared with the normal control group,the expressions of PI3K and AKT1 were significantly in-creased and mTOR mRNA was significantly decreased in the model group(P<0.01,P<0.05). Compared with the mod-el group,the expressions of PI3K and AKT1 were significantly decreased and mTOR mRNA was significantly increased in the sodium houttuyfonate group and dexamethasone group(P<0.01,P<0.05). Compared with the dexamethasone group, the expression of mTOR mRNA was significantly increased in the sodium houttuyfonate group(P<0.05). The pathological observation indicated that there were local pulmonary consolidation and a extensive neutrophil infiltration in the alveolar cav-ity. Prominent pulmonary interstitial fibrous hyperplasia was observed in the model group. The pathological manifestations were much ameliorated than those of the model group,and only mild interstitial pneumonia and a slight fibrous hyperplasia were seen in the sodium houttuyfonate and the dexamethasone groups. Conclusions Sodium houttuyfonate reduces the in-jury of lung tissue and has protective effect on COPD rats. The mechanism is probably related to the down-regulatation of expression of PI3K and AKT1 mRNA and up-regulatation of expression of mTOR mRNA in COPD rats.

Objective To explore the value of transesophageal doppler in transurethral resection prostate.Methods Thirty-six patients (aged 60-85 years, falling into ASA grade Ⅰ-Ⅲ) of benign prostatic hyperplasia for undergoing transurethral resection prostate were enrolled.Through multifunction monitor, CVP, Narcotrend index(NI) were monitored before anesthesia induction (T0), 20 minutes after anesthesia induction (T1), after irrigating fluid of 5 000 ml (T2) and 10 000 ml (T3) and 15 000 ml (T4) and 20 000 ml (T5).By transesophageal doppler, FTc(corrected flowtime), SV(stroke volume), ΔPV(peak flow vaviable quantity) were monitored at T1-T5.Results CVP at T1-T5 were not significantly changed compared with that at T0.Compared with T1, FTc, SV at T3-T5were significantly increased and ΔPV at T2-T5were significantly decreased (P<0.05).CVP levels correlated significantly with both FTc (r=0.702, P<0.01) and SV (r=0.595, P<0.01).CVP negatively correlated significantly with ΔPV (r=-0.351, P<0.05).Furthermore, FTc correlated significantly with the concentrationof Na+(r=0.672,P<0.01).No patient had serum sodium ion concentration less than 125 mmol/L.Conclusion FTc of transesophageal doppler is as accurate as central venous pressure in monitoring hemodynamic changes, and even more sensitive than CVP.It is useful in early diagnosis and treatment of TURS.

Objective To discuss the outcomes of transvaginal repair and transabdominal repair for complex vesicovaginal fistula (VVF).Methods The data of complex VVF patients undergoing surgical repair in Peking University First Hospital were retrospectively collected between January 2009 and December 2016.The surgical modalities for complex VVF included transvaginal repair with layered closure and transabdominal repair with full thick vascular peritoneal interposition.The subtype distribution of complex VVF in transabdominal repair group and transvaginal repair group were recorded.The present study included 63 complex VVF patients with the median age of 46 years (range 26-60 years).There were 32 cases undergoing transvaginal repair with layered closure and 31 cases undergoing transabdominal repair with full thick vascular peritoneal interposition.The proportion of cases having failed previous repairs was significantly higher in transvagical repair group (30/32 vs.23/31,P =0.034).Compared with patients with transvaginal repair,patients with transabdominal repair tended to have multiple VVF without statistic significance (18.8％ vs.29.0％,P =0.338).Patients with transabdominal repair had larger VVF than patients with transvaginal repair (median:1.0cm vs.0.5cm,P ＜ 0.001).Results There were 2 cases suffering from fat liquefaction of surgical incision and 1 case suffering from adhesive intestinal obstruction in patients undergoing transabdominal repair.In the median follow-up duration of 24 months (range 8-102 months)and 29 months (range 8-78 months),the successful rates of transvaginal repair and transabdominal repair were 75％ (24/32) and 93.5％ (29/31).Severe lower urinary tract symptoms occurred in one patient who had urine leakage after transabdomnal repair.The bladder volume of patients in transabdominal group recovered at postoperative 3-6 months.Conclusions In consideration of surgical invasion and fistula condition,transvaginal repair with layered closure and transabdominal repair with full thick vascular peritoneal interposition should be performed individually for complex VVF.Meanwhile,the surgeons need pay attention to other perioperative management.

Objective To investigate the correlation of right ventricular (RV) to left ventricular (LV) volume ratio (RVv/LVv) measured by chest CT with pulmonary arterial pressure estimated by echocardiography before lung transplantation.Methods We reviewed 104 cases of lung transplant.According to the mean pulmonary arterial pressure (mPAP) exceeding 25 mmHg by right heart catheterization,hypertensive group (n =74) and normotensive group (n =30) were set up.Risk factors were assessed individually and adjusted for confounding by a multivariable logistic regression analysis.The area under the curve (AUC) for predicting pulmonary hypertension on chest CT and echocardiography was calculated.Results RVv/LVv and echocardiography-derived ptlmonary arterial systolic pressure (PASP) were significantly different between the two groups (P ＜ 0.05).In the hypertensive group,there was strong correlation between the RVv/LVv and PASP from catheterization (R =0.82,P＜0.001),also between the P ASP from echocardiography and catheterization (R =0.60,P＜ 0.001).The ROC curve displayed that with 0.85 as the cutoff for RVv/LVv,the sensmitivity,specificity and accuracy rate for predicting mPAP over 25 mmHg were 87.5％,91.8％ and 90.9％,respectively.The ROC curve also displayed that based upon an echocardiography-derived PASP of 35 mmHg as the cutoff point,the sensitivity,specificity and accuracy rate for predicting mPAP over 25 mmHg were 91.2％,90.8％ and 88.4％,respectively.Conclusion RV/LV volume ratios on chest CT and echocardiographic evaluation correlate well with PASP assessed by right heart catheterization and can be used to predict pulmonary hypertension with high sensitivity and specificity.

Objective To evaluate the effects of flurbiprofen axetil administered at different time points on oxygenation in the patients undergoing one-lung ventilation (OLV).Methods Ninety patients of both sexes,aged 45-64 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,undergoing elective thoracoscope-assisted pulmonary lobectomy,were assigned into 3 groups (n =30 each) using a random number table:control group (group C),preoperative administration group (group F1) and intraoperative administration group (group F2).Flurbiprofen axetil (10 mg/ml) and fat emulsion 10 ml were injected intravenously at 15 min before operation in F1 and C groups,respectively.Flurbiprofen axetil 10 ml was intravenously injected immediately after the beginning of OLV in group F2.At 15 min before operation (T1),15 and 30 min of OLV (T2,3),and 15 min after restoration of two-lung ventilation (T4),airway peak pressure (Ppeak) and dynamic lung compliance (Cdyn) were recorded,arterial blood samples were collected for blood gas analysis.The arterial oxygen partial pressure (PaO2) was recorded,and the oxygenation index (OI) and intrapulmonary shunt (Qs/Qt) were calculated.The concentrations of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-K-PGF1α) in serum were measured by enzyme-linked immunosorbent assay,and TXB2/6-K-PGF1α ratio was calculated.The development of interrupting OLV due to SpO2＜90％ and postoperative dyspnea,pulmonary infection,atelectasis and length of hospital stay were recorded.Results Compared with group C,PaO2 and OI were significantly increased,and Qs/Qt was decreased at T2,3,the serum concentrations of TXB2 and 6-K-PGF1α were decreased,and TXB2/6-K-PGF1α ratio was increased at T2-4,the incidence of interrupting OLV was decreased (P＜0.05),and no significant change was found in the parameters mentioned above in group F2 (P＞0.05).Compared with group F1,PaO2 and OI were significantly decreased at T2,3,Qs/Qt was increased at T2,and the serum concentrations of TXB2 and 6-K-PGF1α were increased,and TXB2/6-K-PGF1α ratio was decreased at T2-4 in group F2 (P＜0.05).There was no significant difference in the incidence of postoperative dyspnea,pulmonary infection and atelectasis and length of hospital stay between the three groups (P＞0.05).Conclusion Flurbiprofen axetil injected at 15 min before operation can significantly improve oxygenation and prevent the development of hyoxemia in the patients undergoing OLV,however,flurbiprofen axetil administered immediately after the beginning of OLV has no such effect.