Bs 300

Venous blood samples were collected from the forearm after a 12-hour fast according to standard techniques. The samples were collected in the educational centers and later transported in a rigid thermal container with cold packs to a laboratory where the blood serum was separated by centrifugation. Subsequently, the serum samples were analyzed in a single central laboratory in the capital of San Luis Potosí. Total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglyceride, and glucose levels were assessed on the automated analytical platform BS300 (Mindray®, Nanshan, Shenzhen, China) following the internal test protocols and the use of commercially available reagent kits. For the classification of hyperglycemia, levels greater than 100 mg/dl (5.55 mmol/L) were considered the clinical cutoff point, while for prediabetes, levels of ≥100 mg/dl (5.55 mmol/L) to <126 mg/dl (6.99 mmol/L) were considered the cutoff point according to the standards established by international guidelines (18 (link)).

The calcium and phosphorus (P) concentration in serum was evaluated through habitual methods using an automated analyzer (Bs-300, Mindray Biomedical Electronics Co., Ltd., Shenzhen, China).

Clinical chemistry analysis of the urine was carried out on an automatic biochemistry analyzer (Mindray BS-300) using appropriate kits with the following parameters: CRE, UACR. The renal tissues were rapidly dissected, fixed overnight in 10% buffered formalin, and then embedded in paraffin, made into 4 mm sections and further stained with HE and PAS.

Samples were analyzed on the fully automated Mindray BS300 device with the Relassay kit. 300 μL of reagent 1 (measurement buffer) and 45 μL of the sample were mixed, and the first reading was performed at 530 nm after 30 sec. Next, 15 uL of reagent 2 (prochromogenic solution) was added to the mixture, and the second reading was performed at 530 nm after 5 min of incubation. A standard solution containing 10 μmol/L hydrogen peroxide (H₂O₂) (equivalent/liter) supplied in the kit was used for standard measurement. The first and second measurements were performed three times, and their averages were calculated. The absorbance change (ΔAbs) was calculated by subtracting the first absorbance value (A1) from the second absorbance value (A2). The TOS levels were calculated using the formula provided in the kit and expressed as mmol H₂O₂ Eq/L: $\begin{array}{c}\text{TOS}=\frac{\mathrm{\Delta }\text{Abs\hspace{0.17em}sample}}{\mathrm{\Delta }\text{Abs\hspace{0.17em}standard}}\times \text{standard\hspace{0.17em}concentration\hspace{0.17em}}\frac{10\mu \text{mol}}{\text{L}}.\end{array}$

In this study, blood glucose level was measured using the Precision G Blood Glucose Testing System (Abbott Laboratories, Abbott Park, IL). Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed on mice after 10 weeks of bFGF treatment (Figure 1A). After a 12 h fast, mice were orally administered with 50% glucose solution at a dose of 2 g/kg body weight. For ITT, mice were i.p. injected with insulin solution at a dose of 0.35 U/kg body weight after a 4 h fast. Blood glucose levels were measured from the tail vein at time 0, 30, 60, 90, 120, and 150 min. The levels of urea nitrogen (UN), uric acid (UA), urinary albumin to creatinine ratio (UACR) in the urine was determined using an automatic biochemistry analyzer (Mindray BS-300).
For pathologic examination, the kidney tissues were harvested, fixed overnight in 4% paraformaldehyde, and then embedded in paraffin. After deparaffinization and rehydration, the paraffin sections (5 mm) were stained with hematoxylin and eosin (HE) for routine histopathological observations. Periodic acid-Schiff (PAS) and Masson’s trichrome staining were used to determine collagen deposition and fibrosis, respectively.