OBJECTIVE To investigate the feasibility and application of enzyme-linked bridging assay(ELBA)method to the pharmacokinetic evaluation of antisense strand siRNA drug. METHODS Antisense strand RNAs were diluted in LNCap cell lysates from 5 to 50 000 pmol·L-1 to construct the quantification curves. We transfected the intact double-strand siRNA at a final concentration 100 nmol·L-1 targeting Polo-like kinase into the LNCap cells and investigated the specificity of ELBA quantitating the siRNA antisense strand in cell supernatant,cell lysates and RNA-induced silencing complex( RlSC). Quantification curves were constructed and validated in biological matrices such as plasma (5-25 000 pmol·L-1 )and multiple tissues(liver,heart,spleen,and kidneys)(3-6250 pmol·L-1 ). The prostate specific membrane antigen aptamer siRNA delivery system with the intact siRNA concentration of 15 nmol·kg-1 was prepared. The siRNAs were delivered into the LNCap xenogrant tumor model in C57 mice by tail vein injection. The concentration of siRNA antisense strand was determined in plasma and tissues 30 min post administration by ELBA. RESULTS The quantitative range of antisense strand siRNA in cell lysates was 5-50 000 pmol·L-1 ,and ELBA method could quantify the siRNA antisense strand concentration from cell lysates and RlSC in LNCap cells transfected with double-strand siRNA. ln addition,ELBA could specifically reflect the single antisense strand concentration instead of intact siRNA double strands in plasma. The quantification range of siRNA antisense strand using ELBA in plasma was 5-25 000 pmol·L-1 and 3-3125 pmol·L-1 in tissues. About 30 min post administration of PSMA aptamer-siRNA,the antisense strand of siRNA was distributed mainly to the tumor,liver,kidneys,blood and spleen in sequence. The distribution profile might be attributed to the target delivery and siRNA pharma-codynamics. CONCLUSION The ELBA method is successfully applied to the siRNA antisense strand pharmacokinetic evaluation,which provides an alternative for pharmacokinetic studies of siRNA-based drugs.

Objective: To investigate the incidence of renal insufficiency in solitary kidney patients and analyze the risk factors. Methods: Patients with solitary kidney who were admitted to the Second Hospital of Lanzhou University from January 2012 to January 2019 were retrospectively selected as subjects. According to estimated glomerular filtration rate (eGFR) level, the patients were divided into two groups: eGFR<60 ml·min^-1·(1.73 m²)^-1 group and eGFR≥60 ml·min^-1·(1.73 m²)^-1 group. The data of the general information, laboratory examinations and kidney size were collected, and the differences of the above indicators between the two groups were compared. Logistic regression model was used to analyze the related factors of renal function decline. Results: (1) A total of 323 solitary kidney patients with age of (53.8±15.8) years and median duration of 10.0 years were enrolled in the study, including 203 males (62.8%). There were 150 cases (46.4%) with hypertension, 136 cases(42.1%) with proteinuria, and 134 cases (41.5%) with renal insufficiency, even 29 cases(9.0%) had developed into end-stage renal disease. (2) Compared with those in eGFR≥60 ml·min^-1·(1.73 m²)^-1group, patients in eGFR<60 ml·min^-1·(1.73 m²)^-1 group had higher age, mean arterial pressure, serum creatinine, serum uric acid, fasting blood glucose, and higher proportion of hypertension and proteinuria, but had lower proportion of congenital solitary kidney, hemoglobin, plasma albumin and residual kidney diameter. The differences of above indicators were statistically significant (all P<0.05). (3) Logistic regression analysis showed that increasing age (every ten years, OR=1.752, 95% CI 1.455-2.109, P<0.001), anemia (OR=2.327, 95% CI 1.356-3.994, P=0.002), hyperuricemia (OR=5.097, 95% CI 2.873-9.042, P<0.001) and high urine protein level (every 1+, OR=1.515, 95% CI 1.197-1.919, P=0.001) were independent risk factors for renal dysfunction in solitary kidney patients. Conclusions The incidence of renal insufficiency in solitary kidney patients is 41.5%. Patients with solitary kidney may perform varying degrees of kidney damage, such as hypertension, proteinuria and eGFR decline. Increasing age, anemia, hyperuricemia and high urine protein level are independent risk factors for renal insufficiency in solitary kidney patients.

Objective:To explore the incidence and occurrence time of gallstone disease after bariatric surgery.Methods:Retrospectively analyzed the clinical data of 187 patients with morbid obesity who underwent bariatric surgery in the Department of General Surgery, Beijing Friendship Hospital, Capital Medical University from Dec. 2017 to Aug. 2019. All patients did not receive prophylactic ursodeoxycholic acid. All patients were underwent abdominal ultrasound and MRI examination preoperatively, and at least one abdominal ultrasound, MRI examination postoperatively. The incidence and occurrence time of gallstones and biliary sludge in patients with different bariatric surgery were analyzed respectively. Measurement data conforming to the normal distribution were described as mean ± standard deviation ( Mean± SD). Measurement data consistent with skewed distribution were described as median (lower quartile, upper quartile). Counting data were described as a percentage (%). Kruskal-Wallis test was used for comparison among groups, and then Bonferroni correction was used for pairwise comparison. Results:The follow-up time was up to Dec. 31, 2020, with a median follow-up time of 27.0 (22.0, 31.0) months. Thirty-four patients (18.2%, 34/187) developed gallstones after bariatric surgery. Individually, it was 18.0%(30/167) in LSG group, 22.2%(2/9) in LRYGB group, 11.1%(1/9) in LOAGB group and 50.0%(1/2) in LOAGB revisional surgery group. Eighteen patients (9.6%, 18/187) were found biliary sludge formation, among which 8.4% (14/167), 22.2% (2/9), and 22.2% (2/9) underwent LSG, LRYGB, and LOAGB, respectively. The rates of weight loss and BMI loss in patients with postoperative gallstone-formation were 21.4 (18.7, 23.6)% and 21.4 (18.6, 23.5) %, respectively. Three patients (1.6%, 3/187) had newly developed symptomatic gallstones, and all of them underwent LSG. The mean occurrence time for biliary sludge and gallstone was 85.5 (28.8, 98.8) and 103.5 (93.0, 179.3) days, respectively. Statistical difference in occurrence time was only found between postoperative gallstone and biliary sludge formation ( P=0.009). Conclusion:Without drug intervention, the incidence of gallstone after bariatric surgery was about 18.2% (34/187), which requires close clinical attention. Abdominal ultrasonography and the T2WI sequence of upper abdominal MRI can help to diagnose gallstone and monitor its changes.

Objective:To summarize the clinical manifestations and color Doppler ultrasonography of benign lymphoepithelial disease (BLL) in lacrimal gland, so as to improve the diagnostic accuracy of lacrimal BLL.Methods:Clinical manifestations and color Doppler flow imaging (CDFI) features of lacrimal BLL in 16 patients (2 males and 14 females, with 31 lesions) who visited the Second Hospital of Hebei Medical University from November 2014 to August 2019 were retrospectively analyzed.Results:The performance for lesions in patients with duration less than 3 months was scattered in irregular low echo, lesion blood flow signals within the rich (Adler Ⅱ level), pathologic examination results showed more lymphocytes were seen in the lacrimal matrix, and the myoepithelium of the lacrimal duct grew to form the epithelium-myocutaneous island. The ultrasonic feasures in patients with course of 3-6 months were in multiple categories such as circular low echo, "honeycomb" change, CDFI showed lesions with a lot of blood flow signals (Adler Ⅲ level), the pathological examination results indicated that there were a large number of lymphocytes and epithelial-musculocutaneous islands in the lacrimal matrix, and the lymphocytes were significantly increased compared with patients with the course of disease less than 3 months. The ultrasound results in patients with a course of more than 6 months showed lesions in multiple categories such as circular low echo, and large low echo, greater than 3 mm in diameter, CDFI showed lesions within the same large amounts of blood flow signals (Adler Ⅲ level), the pathological examination results were consistent with the course of 3 to 6 months. One patient had positive tuberculin test and 11 had higher IgG4 than normal.The number of lymphocytes increased with the prolongation of disease course.Conclusions:With the expert knowledge of color Doppler ultrasonographic characteristics of lacrimal gland BLL and with the serum IgG4 level helps to improve the accuracy of ultrasonic diagnosis.

Objective:To investigate the expression of microRNA-155 (miR-155) in serum and kidney of C57BLKS/db (db/db) mice and its role in the pathogenesis of diabetic kidney disease (DKD).Methods:The db/db mice ( n=24) were divided into 6, 8, and 10 weeks old groups ( n=8) with age increasing according to the random number table, and C57BL/6 mice of the same age were used as control group. The expression of miR-155 in mouse serum and kidney tissue was determined using real-time quantitative PCR. The mRNA and protein expression of Ets-1, eNOS, AGTR1 in renal tissues was verified by real-time quantitative PCR, Western blotting and immunohistochemistry. Results:Compared with the control group, the expression of miR-155 in serum of db/db mice at 6, 8 and 10 weeks of age were significantly increased (all P<0.01), and the increase of miR-155 was most obvious at 10 weeks of age ( P<0.01). Meanwhile the expression of miR-155 in kidney tissues of 6, 8 and 10 weeks old db/db mice was significantly up-regulated (all P<0.01), and the highest expression of miR-155 was at 10 weeks of age ( P<0.01). Immunohistochemistry showed that Ets-1, eNOS and AGTR1 were localized in glomerular endothelial cells. The results of real-time quantitative PCR showed that the mRNA expression of Ets-1, eNOS and AGTR1 were down-regulated in the kidney tissues of db/db mice at 6, 8 and 10 weeks of age compared to the control(all P<0.05), and the level of down-regulation was the most obvious at 10 week. Western blotting results showed that there was no significant change in Ets-1, eNOS and AGTR1 in 6-week-old db/db mice compared to the control group; the eNOS protein expression was down-regulated at 8 weeks of age ( P<0.05); the expression of AGTR1 protein was down-regulated ( P<0.05), and the protein expression of Ets-1 and eNOS were significantly down-regulated at 10-week age (both P<0.01). Conclusions:The expression of miR-155 in serum and kidney tissues of db/db mice increases during the progression of DKD, while the expression of miR-155 target genes Ets-1, eNOS and AGTR1 decreases with the progression of DKD. MiR-155 may participate in the development and progression of DKD by inhibiting its target genes Ets-1, eNOS and AGTR1, affecting endothelial cell function.

We aimed to investigate the relationship of dietary zinc intake with new-onset hypertension among Chinese adults. A total of 12,177 participants who were free of hypertension at baseline from the China Health and Nutrition Survey were included. Dietary intake was assessed by three consecutive 24-h dietary recalls combined with a household food inventory. Participants with systolic blood pressure ≽ 140 mmHg or diastolic blood pressure ≽ 90 mmHg or diagnosed by a physician or under antihypertensive treatment during the follow-up were defined as having new-onset hypertension. During a median follow-up duration of 6.1 years, 4269 participants developed new-onset hypertension. Overall, the association between dietary zinc intake and new-onset hypertension followed a J-shape (P for non-linearity < 0.001). The risk of new-onset hypertension significantly decreased with the increment of dietary zinc intake (per mg/day: hazard ratio (HR) 0.93; 95% confidence interval (CI) 0.88-0.98) in participants with zinc intake < 10.9 mg/day, and increased with the increment of zinc intake (per mg/day: HR 1.14; 95% CI 1.11-1.16) in participants with zinc intake ≽ 10.9 mg/day. In conclusion, there was a J-shaped association between dietary zinc intake and new-onset hypertension in general Chinese adults, with an inflection point at about 10.9 mg/day.
Adult ; Humans ; Cohort Studies ; Zinc ; Diet ; Hypertension/epidemiology* ; Eating ; China/epidemiology*