Discrimination of Liver Metastases of Digestive System Neuroendocrine Tumors From Neuroendocrine Carcinoma by Computed Tomography-Based Radiomics Analysis.

OBJECTIVE: The aim of the study is to investigate the value of computed tomography (CT) radiomics features to discriminate the liver metastases (LMs) of digestive system neuroendocrine tumors (NETs) from neuroendocrine carcinoma (NECs). METHODS: Ninety-nine patients with LMs of digestive system neuroendocrine neoplasms from 2 institutions were included. Radiomics features were extracted from the portal venous phase CT images by the Pyradiomics and then selected by using the t test, Pearson correlation analysis, and least absolute shrinkage and selection operator method. The radiomics score (Rad score) for each patient was constructed by linear combination of the selected radiomics features. The radiological model was constructed by radiological features using the multivariable logistic regression. Then, the combined model was constructed by combining Rad score and the radiological model into logistic regression. The performance of all models was evaluated by the receiver operating characteristic curves with the area under curve (AUC). RESULTS: In the radiological model, only the enhancement degree (odds ratio, 8.299; 95% confidence interval, 2.070-32.703; P = 0.003) was an independent predictor for discriminating the LMs of digestive system NETs from those of NECs. The combined model constructed by the Rad score in combination with the enhancement degree showed good discrimination performance, with AUCs of 0.893, 0.841, and 0.740 in the training, testing, and external validation groups, respectively. In addition, it performed better than radiological model in the training and testing groups (AUC, 0.893 vs 0.726; AUC, 0.841 vs 0.621). CONCLUSIONS: The CT radiomics might be useful for discrimination LMs of digestive system NECs from NETs.

[Clinical research of acupuncture therapy combined with domperidone in treatment of diabetic gastroparesis of liver stagnation and spleen deficiency].

OBJECTIVE: To observe the clinical efficacy of shuanggu yitong acupuncture therapy (the therapy for both replenishment and unblocking) combined with domperidone on diabetic gastroparesis (DGP) of liver stagnation and spleen deficiency pattern and explore its effect mechanism. METHODS: DGP patients differentiated as liver stagnation and spleen deficiency pattern were divided into a control group (n=42) and an observation group (n=42) according to the random number table. The patients in the control group took domperidone tablets orally, 10 mg each time, 3 times a day for 28 days. In the observation group, on the base of the treatment as the control group, shuanggu yitong acupuncture therapy was applied to Baihui (GV20), Shenting (GV24), Zhongwan (CV12), bilateral Zusanli (ST36), Hegu (LI4)and Taichong (LR3), stimulated for 30 min in each treatment. Acupuncture was given once daily, 3 times a weeks for 28 days consecutively. Fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin (HbA1c) were detected before and after treatment in the patients of two groups separately. The score of symptom severity index of gastroparesis (GCSI), traditional Chinese medicine (TCM) syndrome score and gastric emptying rate were assessed in the patients of two groups. Using ELISA, radioimmunoassay and colorimentry methods, the contents of motilin in plasma, gastrin, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and interferon-gamma (INF-γ) in serum, as well as the activity of superoxide dismutase (SOD), reactive oxygen species (ROS) and malondialdehyde (MDA) in the serum were determined in the two groups. The clinical curative effect was evaluated. RESULTS: After treatment, the levels of FBG, 2 h PBG and HbA1c, the scores of GCSI and TCM syndrome, the contents of motilin in plasma, gastrin, TNF-α and MDA, as well as the activity of ROS in serum were all reduced when compared with those before treatment in each group (P<0.05, P<0.01), while gastric emptying rate and SOD activity in the serum were higher than those before treatment (P<0.05, P<0.01). After treatment, the serological content of INF-γ was lower than that before treatment in the control group (P<0.05), and the contents of IL-6 and IL-1ß were reduced than those before treatment in the observation group (P<0.05). Compared with the control group, the levels of FBG, 2 h PBG and HbA1c, the scores of GCSI and TCM symptoms, the contents of motilin in plasma, gastrin, TNF-α, MDA, IL-6 and IL-1ß, and the activity of ROS in serum in the observation group were all lower significantly (P<0.05, P<0.01), while the SOD activity and gastric emptying rate in the observation group were higher than those in the control group (P<0.05, P<0.01). The total effective rate was 90.5% (38/42) in the observation group, better than the control group (73.8%, 31/42, P<0.05). CONCLUSION: Shuanggu yitong acupuncture therapy combined with domperidone remarkably relieves the clinical symptoms and improves the gastric emptying rate, effectively reduces motilin and gastrin and regulates oxidative stress and inflammatory responses in the patients with DGP of liver stagnation and spleen deficiency.

[Clinical efficacy and time-effect relationship of dynamic qi acupuncture for acute lumbar sprain].

OBJECTIVE: To observe the clinical efficacy of dynamic qi acupuncture for acute lumbar sprain, and to explore the differences of different needle retention time on the improvement of pain, lumbar mobility and lumbar dysfunction. METHODS: A total of 160 patients with acute lumbar sprain were randomly divided into an observation group A (40 cases, 4 cases dropped off), an observation group B (40 cases, 2 cases dropped off), an observation group C (40 cases, 4 cases dropped off) and a medication group (40 cases, 6 cases dropped off). The patients in the observation group A, the observation group B and the observation group C were treated with acupuncture at "lumbago point" and Sanjian (LI 3) on the left side, and during the needles were kept for 10, 20 and 30 min respectively, the patients were required to take tolerable lumbar active activities, once a day; the patients in the medication group were treated with celecoxib capsules, 0.2 g each time, twice a day. All the patients were treated for 5 d. Before and after treatment, the scores of numerical rating scale-11 (NRS-11), lumbar range of motion (ROM) and modified Oswestry disability index (ODI) were observed, and the clinical efficacy of each group was evaluated. RESULTS: After treatment, the scores of NRS-11, ROM and ODI in each group were decreased compared before treatment (P<0.01). The decreased degree of NRS-11, ROM and ODI in each observation group was greater than that in the medication group (P<0.05), and the decreased degree of ROM and ODI in the observation group B and the observation group C was greater than that in the observation group A (P<0.05). The total effective rates were 94.4% (34/36) in the observation group A, 94.7% (36/38) in the observation group B and 97.2% (35/36) in the observation group C, respectively, which were higher than 79.4% (27/34) in the medication group (P<0.05). CONCLUSION: Dynamic qi acupuncture with needle retention for 10, 20 and 30 min all could effectively improve the pain, lumbar mobility and lumbar dysfunction in patients with acute lumbar sprain. If the lumbar dysfunction is severe, needle retention for 20 min or more is recommended.

Qualitative and quantitative parameters on hepatobiliary phase of gadoxetic acid-enhanced MR imaging for predicting pathological response to preoperative systemic therapy in colorectal liver metastases.

PURPOSE: To explore the value of parameters of the on hepatobiliary phase (HBP) of pre-treatment gadoxetic acid-enhanced magnetic resonance imaging (MRI) for predicting pathological response to systemic therapy in colorectal liver metastases (CRLMs), compared with response evaluation criteria in solid tumors, version 1.1 (RECIST 1.1). METHODS: A total of 96 patients with CRLMs who underwent gadoxetic acid-enhanced MRI prior to treatment and then liver resection from January 2017 to December 2021 were enrolled. The pathological response was assessed by the percentage of residual tumors (RTs), and CRLMs were classified into two groups according to the pathological response grade (PRG): (1) strong response (including PRG2 and PRG3, RTs ≤ 10%), and weak response (PRG1, RTs > 10%). Two radiologists evaluated the enhancement pattern and degree of CRLMs on the HBP. The diameter, mean and standard deviation (SD) value of signal intensity (SI) of CRLMs on pre-contrast and HBP images were recorded. Relative tumor enhancement (RTE) and the SD ratio (SDR) were calculated. These parameters were analyzed in terms of pathological response on a lesion-by-lesion basis. RESULTS: Totally, 263 CRLMs were classified into: the strong response group (PRG2, n = 57; PRG3, n = 7) and the weak response group (PRG1, n = 199). RTE and SDR values were significantly higher in the strong response group than in the weak response group (P < 0.001). RTE values (P < 0.001) and SDR values (P = 0.031) were independent factors for predicting strong response. The area under curve (AUC) of RTE and SDR values were 0.725 and 0.652, respectively. The combination of these parameters was 0.750, which performed better than RECIST 1.1 (0.750 vs 0.531; P < 0.001). CONCLUSIONS: RTE and SDR values on HBP are potential features in predicting pathological response to systemic therapy in CRLMs.

[Protective effect and mechanism of electroacupuncture of "Biao-Ben" acupoints combination for mitochondrial dysfunction in diabetic nephropathy rats].

OBJECTIVE: To study the protective effect of electroacupuncture (EA) at "Biao"-acupoints, "Fenglong"(ST40) and "Zhongwan"(CV12), used for treating symptoms of the disease, and "Ben"-acupoints, "Zusanli"(ST36) and "Guanyuan"(CV4), for treating the root cause of the disease on oxidative stress injury of renal mitochondria through SIRT1/PGC-1α signal pathway in rats with diabetic nephropathy (DN). METHODS: A total of 33 male Wistar rats were randomized into normal (n=10), model (n=12) and EA (n=11) groups.The DN model was established by feeding the rats with high-sugar and high-fat diet for 6 weeks combined with streptozotocin (STZ, 35 mg/kg, i.p.). EA (4 Hz/60 Hz, 1 mA)was applied to ST36-ST40 and CV4-CV12 for 15 min, once every other day for 8 weeks. The rats' body weight was recorded, urine in 24 hours (24-h UP) was collected to measure the urine protein level, and the fasting blood glucose (FBG) level detected by using a glucometer. The levels of serum glycosylated hemoglobin (HbA1c), creatinine (Scr) and urea nitrogen (BUN) were assayed using immunoturbidi-metry, picric acid method and urease method, respectively, and those of serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) detected using an automatic biochemical analyzer. The kidney tissue was collected for assaying the activity of superoxide dismutase (SOD) with xanthine oxidase method, glutathione (GSH) activity with dithio-dinitrobenzoic acid method, catalase （CAT） activity with ammonium molybdate spectrometric method, and malondialdehyde (MDA) content with thiobarbituric acid method. Histopathological changes of the kidney tissue were observed by microscope after hematoxylin-eosin staining (HE), periodate Schiff staining (PAS) and Masson staining, separately, and its subcellular structure was observed under transmission electron microscopy. The expression levels of renal SIRT1 and PGC-1α mRNAs and proteins were detected by quantitative real-time PCR and Western blot, and the immunoactivity of renal α-smooth muscle actin (α-SMA), and immunofluorescence density of renal collagen Ⅰ (Col Ⅰ), collagen Ⅳ(Col Ⅳ) and fibronec-tin (FN) detected by immunohistochemistry and immunofluorescence assay, respectively. RESULTS: Compared with the normal group, the levels of FBG, HbA1c, BUN, Scr, 24-h UP, TG, TC, LDL-C, MDA, α-SMA, Col Ⅰ, Col Ⅳ and FN proteins were significantly increased (P<0.01), and the levels of body weight, HDL-C, SOD, GSH, CAT, SIRT1 and PGC-1α mRNAs and proteins were decreased in the model group (P<0.01, P<0.05). In comparison with the model group, the levels of FBG, HbA1c, BUN, Scr, 24-h UP, TG, TC, LDL-C and MDA, and the expressions of α-SMA, Col Ⅰ, Col Ⅳ and FN proteins were markedly down-regulated (P<0.05, P<0.01), and those of body weight, HDL-C, SOD, GSH, CAT, and the expressions of SIRT1 and PGC-1α mRNAs and proteins significantly up-regulated (P<0.01, P<0.05) in the EA group. HE staining showed mesangial dilatation, glomerular hypertrophy and mesangial matrix accumulation; PAS staining showed an increase of the glomerular extracellular matrix deposition; and Masson staining displayed an enhancement of glomerular fibrosis and interstitial space expansion; and electron microscope revealed foot process fusion, basement membrane thickening and organelle injury in the rat's kidney of the model group, which were relatively milder in the EA group. CONCLUSION: EA of ST36-ST40 and CV4-CV12, "Biao-Ben" acupoints combination, can alleviate oxidative stress and mitochondrial dysfunction in DN rats, which may be associated with its functions in up-regulating SIRT1/PGC-1α signaling, and decreasing renal fibrosis.

[Electroacupuncture preconditioning alleviates renal injury and oxidative stress through Sirt3/MnSOD pathway in type 2 diabetic rats].

OBJECTIVE: To explore the protective effect and molecular mechanism of electroacupuncture (EA) preconditioning on renal injury in type 2 diabetic rats. METHODS: Fifty male Wistar rats were randomly divided into control, model, EA, EA+inhibitor, and inhibitor groups, with 10 rats in each group. Diabetes model was established by high fat and high glucose diet and intraperitoneal injection of streptozotocin (40 mg/kg). EA (2 Hz, 1 mA) preconditioning was applied to "Guanyuan" (CV4), "Zhongwan" (CV12), bilateral "Zusanli" (ST36) and "Fenglong" (ST40) for 15 min, once every other day for 8 weeks. Rats of the inhibitor and EA+inhibitor groups were given intraperitoneal injection of 3-TYP (50 mg/kg) once every other day for a total of 3 times. The body weight, kidney mass, and renal index were recorded. The contents of urine microalbumin (ALB), 24 h urine 8-hydroxydeoxyguanosine (8-OHdG) and activities of reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), glutathione glycine peroxidase (GSH-Px) in kidney were detected by ELISA. The activities of mitochondrial respiratory chain enzyme complex (RCCⅠ-RCCⅣ) in kidney were detected using spectrophotometric method. HE staining, Masson staining and transmission electron microscopy were used to observe the changes of renal structure. The protein and mRNA expressions of silent information regulator 3 (Sirt3), manganese superoxide dismutase (MnSOD) in kidney were detected by Western blot and quantitative real-time PCR, respectively. RESULTS: After modeling and compared with the control group, the contents of ALB, the renal index, activity of ROS and content of 8-OHdG, and the renal collagen volume fraction (CVF) were increased (P<0.01), while the activities of SOD, CAT and GSH-Px, RCCⅠ-RCCⅣ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.01). After the treatment and compared with the model group, the contents of ALB, the renal index, ROS, 8-OHdG, and the CVF were decreased in the EA group(P<0.01, P<0.05), while the activities of SOD, CAT, GSH-Px, RCCⅠ-RCCⅣ, and Sirt3 and MnSOD expression levels were increased (P<0.01, P<0.05)；the RCCⅡ activity and the expression level of MnSOD mRNA were increased (P<0.05) in the EA+inhibitor group; the ALB and 8-OHdG contents and the CVF in the inhibitor group were increased (P<0.05), while the activity of SOD, and Sirt3 and MnSOD expression levels were decreased (P<0.05). In comparison with the EA group, the contents of ALB, the renal index, activities of ROS and 8-OHdG contents, and the CVF were increased (P<0.05, P<0.01), activities of SOD, CAT and GSH-Px and RCCⅠ and RCCⅡ, and the mRNA and protein expressions of Sirt3 and MnSOD were decreased (P<0.05, P<0.01) in both EA+inhibitor group and inhibitor group, whereas the activities of RCCⅢ and RCCⅣ were decreased in the inhibitor group (P<0.05). The therapeutic effect of inhibitor was notably inferior to that of EA+inhibitor in decreasing ALB and 8-OHdG contents, and CVF (P<0.01), and in up-regulating SOD and RCCⅡ activities, Sirt3 and MnSOD expression levels (P<0.01, P<0.05). CONCLUSION: EA preconditioning can increase the expressions of renal Sirt3 and MnSOD in type 2 diabetic rats, thereby reducing the oxidative stress response and protecting the kidneys.

[Clinical observation on filiform-fire needling of "Biaoben acupoint combination" for 33 cases of sequelae of coronavirus disease 2019 during recovery period].

OBJECTIVE: To observe the clinical therapeutic effect of filiform-fire needling of "Biaoben acupoint combination" on the sequelae of patients with coronavirus disease 2019 (COVID-19) during the recovery period. METHODS: A total of 33 patients with COVID-19 during the recovery period were treated with filiform-fire needling at the acupoints of Mingmen (GV 4), Shenzhu (GV 12), Gaohuang (BL 43), Zusanli (ST 36) and Shangjuxu (ST 37), etc., once every other day, 3 times a week, and 3 times was one course of treatment and totally 2 courses of treatment were required. The TCM symptom, Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD) scores, pulmonary function indexes (forced vital capacity [FVC], forced expiratory volume in one second [FEV1], peak expiratory flow [PEF]) and chest CT imaging change were observed before and after treatment, and the therapeutic effect was evaluated. RESULTS: After treatment, the scores of TCM symptom, HAMA and HAMD were decreased compared with those before treatment (P<0.05), and the levels of FVC, FEV1 and PEF were increased compared with those before treatment (P<0.05), and the recovery rate of 22 patients with pulmonary ventilation dysfunction was 86.4% (19/22). After treatment, the lung shadow area was smaller than that before treatment (P<0.05). The effective rate of 25 patients with lung CT abnormalities was 84.0% (21/25). After treatment, 23 cases were cured, 5 cases were markedly effective, 4 cases were effective, 1 case was ineffective, the cured and markedly effective rate was 84.8%. CONCLUSION: The filiform-fire needling of "Biaoben acupoint combination" could significantly reduce the sequelae of cough, fatigue, chest tightness, etc. and mental symptoms such as anxiety and depression in patients with COVID-19 during the recovery period, and promote inflammatory exudation absorption of pulmonary lesion and improve lung ventilation function.

[Electroacupuncture of "Biaoben acupoints" relieves kidney injury by suppressing oxidative stress in diabetic nephropathy rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of combined "Biao"- and "Ben"-acupoint (for treating symptoms and root causes of the disease, respectively) on the expression of kidney forkhead box O1 (FoxO1) and peroxi-some proliferator-activated receptor-γ coactivator-1α (PGC-1α) in diabetic nephropathy (DN) rats, so as to explore its potential mechanisms underlying improvement of DN. METHODS: Wistar rats were randomly divided into normal control (n=10), DN model (n=12), EA (n=11), EA+inhibitor (AS1842856 targeting FoxO1, n=11) and inhibitor (n=11) groups. The DN model was established by high fat and high glucose diet for 6 weeks and intraperitoneal injection of streptozotocin (55 mg/kg). EA (2 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Guanyuan"(CV4), "Fenglong" (ST40) and "Zhongwan"(CV12) for 15 min, once every other day for 8 weeks. The body mass was recorded, and blood glucose detected. The serum was sampled for detecting creatinine (Scr) content with Jaffe's assay, urea nitrogen (BUN) content with urease method. Urine albumin (ALB) and renal reactive oxygen species (ROS) contents were detected with ELISA, renal superoxide dismutase (SOD) activity with xanthine oxidase method, and renal malondialdehyde (MDA) content with thiobarbituric acid method. The renal subcellular structure was observed under transmission electron microscopy, and the expression levels of PGC-1α and FoxO1 proteins in the kidney tissue were detected using Western blot. RESULTS: Compared with the normal control group, the levels of body mass, SOD activity, and FoxO1 and PGC-1α protein expression were significantly reduced (P<0.01), while the contents of blood glucose, and serum Scr and BUN, urine ALB, renal MDA and ROS levels significantly increased in the model group (P<0.01). In comparison with the model group, the levels of body mass, SOD activity, and FoxO1 and PGC-1α expression were significantly increased in the three treatment groups except SOD, expression of FoxO1 and PGC-1α in the inhibitor group (P<0.01, P<0.05), and the contents of blood glucose, Scr, BUN, ALB, MDA and ROS were obviously decreased in the three treatment groups except ALB and ROS in the inhibitor group (P<0.01, P<0.05). The therapeutic effect of EA was notably superior to that of EA+inhibitor and inhibitor in increasing body mass, SOD activity, and FoxO1 and PGC-1α expression levels (P<0.05, P<0.01), and in down-regulating blood glucose, BUN, ALB and ROS levels (P<0.05, P<0.01), suggesting a reduction of the therapeutic effect of EA after administration of the inhibitor AS1842856 of FoxO1. Results of electron microscopy showed diffusely thickened and vague basement membrane, increased mesangial matrix, fused foot process, and reduced volume of endothelial cells with pykno-tic nucleus of the kidney tissue in the model group, which was obviously milder in both EA and EA+inhibitor groups particularly in the EA group. CONCLUSION: EA increases the expression of FoxO1 and PGC-1α in the kidneys of DN rats, thereby reducing the oxidative stress response and protecting the kidneys.

[Thumb-tack needles based on "Biaoben acupoint compatibility" for sequela of COVID-19 during recovery period].

OBJECTIVE: To observe the effect of thumb-tack needles based on "Biaoben acupoint compatibility" on sequela symptoms, mental state and pulmonary ventilation function in patients with coronavirus disease 2019 (COVID-19) during recovery period. METHODS: Fifty cases of COVID-19 during recovery period were randomly divided into an observation group and a control group, 25 cases in each group. The patients in the observation group were treated with thumb-tack needles at Guanyuan (CV 4), Zusanli (ST 36) and Taiyuan (LU 9). The patients in the control group were treated with sham thumb-tack needles at identical acupoints as the observation group. The treatment in the two groups was given once a day, 7-day treatment was taken as a course of treatment, and totally two courses of treatment were given. The TCM symptom score, Hamilton anxiety scale (HAMA) score, Hamilton depression scale (HAMD) score, pulmonary function (forced vital capacity [FVC], forced expiratory volume in the first second [FEV1], peak expiratory flow [PEF]), the severity of pulmonary ventilation dysfunction and pulmonary imaging changes in the two groups were compared before and after treatment. RESULTS: Compared before treatment, the total scores and each item scores of TCM symptom scale, HAMA scores and HAMD scores in the two groups were reduced after treatment (P<0.05). Except for the symptom scores of dry throat and dry stool, the total score and each item score of TCM symptom scale, HAMA score and HAMD score in the observation group were lower than those in the control group (P<0.05). Compared before treatment, FVC, FEV1 and PEF in the two groups were increased after treatment (P<0.05), and those in the observation group were higher than the control group (P<0.05). The severity of pulmonary ventilation dysfunction in the two groups was reduced after treatment (P<0.05), and the severity in the observation group was better than that in the control group (P<0.05). After treatment, the lung shadow area in the two groups was decreased (P<0.05), and that in the observation group was smaller than the control group (P<0.05). The improvement of imaging change in the observation group was better than that in the control group (P<0.05). CONCLUSION: The thumb-tack needles based on "Biaoben acupoint compatibility" could significantly reduce the sequela symptoms, anxiety and depression in patients with COVID-19 during recovery stage, and improve the pulmonary ventilation function.

[Effect of electroacupuncture of "Biao-Ben" acupoints on renal function and hemorheology and eNOS level in patients with early diabetic nephropathy].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Biao-Ben acupoints" (Biao indicates pathogenic factors of disease; Ben refers to body constitution) on renal function, hemorheology and endothelial nitric oxide synthase (eNOS) level in patients with early diabetic kidney disease (DKD), so as to explore its mechanism underlying relieving early DKD. METHODS: A total of 120 patients with early DKD were selected and randomized into 3 groups: medication, conventional acupoints, and "Biao-Ben"acupoints groups by stratified randomization method, with 40 cases in each group. Patients of the me-dication group were treated by routine symptomatic supportive treatment (gleziquantel tablets or subcutaneous injection of insulin \[for hyperglycemia\], candesartan tablet \[hypertension\], simvastatin tablets \[hyperlipidemia\], etc.).Based on the medication group, patients of the conventional acupoint group were treated by EA of bilateral Feishu (BL13), Weiwanxiashu (EX-B3), Weishu (BL21), Shenshu (BL23), Sanyinjiao (SP6), Taixi (KI3) (main acupoints), etc., and those of the Biao-Ben acupoint group treated by EA of main acupoints Zhongwan (CV12), Fenglong (ST40), Xuehai (SP10) and Taichong (LR3) (Biao acupoints), and Guanyuan (CV4) and Zusanli (ST36) (Ben acupoints). The EA treatment was conducted one daily, 5 days a week for 8 weeks. The urine microprotein level in 24 h was detected using an automatic specific protein analyzer, followed by calcula-ting the urine albumin excretion rate (UAER). The serum creatinine (Scr), urea nitrogen (BUN) and cystatin (CysC) contents were detected by using an automatic biochemical analyzer, and the whole blood low-cut viscosity (ηbL), whole blood mid-cut viscosity (ηbM), whole blood high-cut viscosity (ηbH), plasma viscosity (ηp) and fibrinogen (FIB) levels were detected using an automatic hemorheology tester, and the serum eNOS and nitric oxide (NO) levels assayed using enzyme linked immunosorbent assay. The total clinical effective rates were compared and the adverse reactions of the treatment were recorded. RESULTS: Compared with the values before the treatment in each group, the levels of UAER, Scr, BUN, CysC, ηbL, ηbM, ηbH, ηp and FIB were all significantly decreased (P<0.01), while serum eNOS and NO levels significantly increased in the three groups after the treatment (P<0.01). Compared with the medication group, the levels of UAER, Scr, BUN, CysC, ηbL, ηbM, ηbH, ηp and FIB were notably lower (P<0.01, P<0.05), and serum eNOS and NO contents obviously higher in both the conventional acupoint and "Biao-Ben" acupoint groups (P<0.01). Comparison between the two EA groups showed that the levels of UAER, BUN, ηbL, ηbM, ηbH and ηp were lower (P<0.05), whereas the serum eNOS and NO contents were considerably higher (P<0.05) in the "Biao-Ben" acupoint group than in the conventional acupoint group. After the treatment, the total clinical effective rate of the "Biao-Ben" acupoint group was 89.74%(35/39), being significantly higher than those of both the conventional acupoint group (71.05%, 27/38,P<0.05) and medication group (64.10%, 25/39, P<0.05). CONCLUSION: EA of "Biao-Ben" acupoints can improve renal function and reduce microcirculation disorders in patients with early DKD, which may be related to its function in up-regulating the levels of serum eNOS and NO.

[Thumb-tack needle based on Biaoben Genjie theory for dry eye: a randomized controlled trial].

OBJECTIVE: To compare the clinical efficacy of thumb-tack needle and sodium hyaluronate eye drops for dry eye. METHODS: A total of 60 patients with dry eye were randomly divided into a thumb-tack needle group and a western medication group, 30 cases in each group. Based on Biaoben Genjie theory, thumb-tack needle was applied at Zhiyin (BL 67), Lidui (ST 45), Fuyang (BL 59), Yanglao (SI 6), Jingming (BL 1) and Cuanzhu (BL 2) in the thumb-tack needle group, 2 times a week. In the western medication group, 0.1% sodium hyaluronate eye drops were applied, 3 times a day. Both groups were treated for 4 weeks. Before and after treatment, TCM symptom score, tear film break-up time (BUT), SchirmerⅠtest (SⅠT), corneal fluorescein sodium staining (FL) score and 36-item short form health survey (SF-36) score were observed, and the clinical efficacy was evaluated in the two groups. RESULTS: After treatment, the total scores and each item score of TCM symptom in the two groups were decreased compared with before treatment (P<0.05); except for score of visual fatigue and photophobia, the total score and each item score of TCM symptom in the thumb-tack needle group were lower than the western medication group (P<0.05). After treatment, the BUT, SⅠT and SF-36 scores were increased and the FL scores were decreased in the two groups (P<0.05); the BUT, SⅠT and SF-36 score were higher than the western medication group (P<0.05), and the FL score was lower than the western medication group (P<0.05) in the thumb-tack needle group. The total effective rate in the thumb-tack needle group was 93.3% (28/30), which was higher than 80.0% (24/30) in the western medication group (P<0.05). CONCLUSION: Thumb-tack needle based on Biaoben Genjie theory could effectively relieve clinical symptoms of dry eye patients, prolong tear film break-up time, increase tear secretion, improve tear film function and quality of life, and its curative effect is better than sodium hyaluronate eye drops.

NLRP3 ablation enhances tolerance in heat stroke pathology by inhibiting IL-1ß-mediated neuroinflammation.

BACKGROUND: Patients with prior illness are more vulnerable to heat stroke-induced injury, but the underlying mechanism is unknown. Recent studies suggested that NLRP3 inflammasome played an important role in the pathophysiology of heat stroke. METHODS: In this study, we used a classic animal heat stroke model. Prior infection was mimicked by using lipopolysaccharide (LPS) or lipoteichoic acid (LTA) injection before heat stroke (LPS/LTA 1 mg/kg). Mice survival analysis curve and core temperature (TC) elevation curve were produced. NLRP3 inflammasome activation was measured by using real-time PCR and Western blot. Mice hypothalamus was dissected and neuroinflammation level was measured. To further demonstrate the role of NLRP3 inflammasome, Nlrp3 knockout mice were used. In addition, IL-1ß neutralizing antibody was injected to test potential therapeutic effect on heat stroke. RESULTS: Prior infection simulated by LPS/LTA injection resulted in latent inflammation status presented by high levels of cytokines in peripheral serum. However, LPS/LTA failed to cause any change in animal survival rate or body temperature. In the absence of LPS/LTA, heat treatment induced heat stroke and animal death without significant systemic or neuroinflammation. Despite a decreased level of IL-1ß in hypothalamus, Nlrp3 knockout mice demonstrated no survival advantage under mere heat exposure. In animals with prior infection, their heat tolerance was severely impaired and NLRP3 inflammasome induced neuroinflammation was detected. The use of Nlrp3 knockout mice enhanced heat tolerance and alleviated heat stroke-induced death by reducing mice hypothalamus IL-1ß production with prior infection condition. Furthermore, IL-1ß neutralizing antibody injection significantly extended endotoxemic mice survival under heat stroke. CONCLUSIONS: Based on the above results, NLRP3/IL-1ß induced neuroinflammation might be an important mechanistic factor in heat stroke pathology, especially with prior infection. IL-1ß may serve as a biomarker for heat stroke severity and potential therapeutic method.

Chronic preclinical safety evaluation of EPO-018B, a pegylated peptidic erythropoiesis-stimulating agent in monkeys and rats.

EPO-018B, a synthetic peptide-based erythropoiesis stimulating agent (ESA), is mainly designed for treatment of anemia caused by chronic renal failure and chemotherapy against cancer. It overcomes the deficiencies of currently approved ESA, including the frequent administration of temperature-sensitive recombinant protein and anti-EPO antibody-mediated pure red cell aplasia (PRCA). This study was designed to evaluate the potential chronic toxicity of EPO-018B. Subcutaneous administration doses were designed as 0, 0.2, 1 and 10mg/kg for six months for 160 rats (20/gender/group) and 0, 0.3, 3 and 20mg/kg for nine months for 32 monkeys (4/gender/group) once every three weeks. The vehicles received the same volume of physiological saline injection. All animals survived to the scheduled necropsies after six weeks (for rats) and fourteen weeks (for monkeys) recovery period, except for the two high-dose female rats and two high-dose male monkeys, which were considered related to the increased RBCs, chronic blood hyperviscosity and chronic cardiac injury. EPO-018B is supposed to be subcutaneously injected once every month and the intended human therapeutic dose is 0.025mg/kg. The study findings at 0.2mg/kg for rats and 0.3mg/kg for monkeys were considered to be the study NOAEL (the no observed adverse effect level), which were more than ten times the intended human therapeutic dose. Higher doses caused adverse effects related to the liver toxicity, cardiotoxicity, appearance of neutralizing antibodies of EPO-018B and the decrease of serum glucose and cholesterol. Most treatment-induced effects were reversible or revealed ongoing recovery upon the discontinuation of treatment. The sequelae occurred in rats and monkeys were considered secondary to exaggerated pharmacology and would less likely occur in the intended patient population. As to the differences between human beings and animals, the safety of EPO-018B need to be further confirmed in the future clinical studies.

Purification and characterization of a urethanase from Penicillium variabile.

Urethanase produced by Penicillium variabile was purified through ultrasonication, concentration by polyethylene glycol 20,000, and Superdex G-200 gel filtration chromatography. The molecular weight of urethanase was determined to be around 96 kDa by gel filtration. The purified enzyme showed a single band in SDS-PAGE with the molecular weight of ~13.7 kDa, which suggests that the enzyme has a multimeric structure composed of the same subunits. Peptide map fingerprinting analysis was then carried out by MALDI/TOF-TOF MS. Within the known sequences in NCBI, glucosamine-6-phosphate deaminase and 6-phosphogluconate dehydrogenase get high score as compared with urethanase. Sequence analysis informs that N-terminal sequence of urethanase is GTNTADNDAA. The Minchaelis constant (Km) and maximum reaction rate (Vm) of urethanase are 27.2 mmol/L and 156.25 µmol/L min, respectively.

Isolation and characterization of urethanase from Penicillium variabile and its application to reduce ethyl carbamate contamination in Chinese rice wine.

A strain with urethanase activity was isolated from mouse gastrointestine. By combination of morphological characterization of the colony, hyphae, and spore and the sequence analysis of its rDNA ITS, the strain was determined as Penicillium variabile and named as P. variabile JN-A525. The enzymatic properties of urethanase from P. variabile JN-A525 were further studied. The optimum temperature and pH value of urethanase are of 50 °C and 6.0, respectively. The enzyme maintains stability when the temperature is below 50 °C and the pH is in the range of 7.0-10.0. The enzyme also exhibits ethanol tolerance. It can remove ethyl carbamate from Chinese rice wine without the change of flavor substances in the wine.