OBJECTIVETo analyze the fungal composition in Massa Medicata Fermentata based on culture dependent method and independent PCR-SSCP technique.

METHODFungi were directly isolated from Massa Medicata Fermentata samples. The obtained strains were identified according to morphology and DNA sequence. Meanwhile the total fungal DNA was extracted from Massa Medicata Fermentata samples, the cultural independent PCR-SSCP technique based on β-tubulin gene were used to identify the mycobiota.

RESULTAccording to cultural method, Aspergillus flavus and Rhizopus oryzae were present in Massa Medicata Fermentata samples, while A. flavus and A. niger were present in fried Massa Medicata Fermentata samples. In contrast, 5 species were obtained by PCR-SSCP technique, A. flavus was overlapped with fungal taxa derived from culture dependent method; A. ambiguu and A. s ivoriensis were dominant with relative abundance of 57% and 35% respectively, while the relative abundance of A. flavus was as low as 4%. None species was obtained from fried Massa Medicata Fermentata samples.

CONCLUSIONPCR-SSCP based on β-tubulin gene could distinguish fungi into species, culture dependent method combined with culture independent method could better understand the fungal composition associated with Massa Medicata Fermentata fermentation.

Fermentation ; Fungi ; isolation & purification ; Medicine, Chinese Traditional ; Polymerase Chain Reaction ; methods ; Polymorphism, Single-Stranded Conformational ; Tubulin ; genetics

At present, the occurrence and development of cataract in patients operated by vitrectomy is more common. The patient, who would be in cataract surgery, is prone to suffering from higher risks and complications in eyes if he had vitrectomy before than those without vitrectomy. Consequently, it is a significant challenge for ophthalmologists to determine the choice of surgery and the operation during surgery. Compared with cataract surgery after vitrectomy without perfusion, phacoemulsification with 25-gauge perfusion is an effective method, which can maintain the intraocular pressure, stabilize the anterior chamber, and reduce the intraoperative and postoperative complications. In conclusion, phacoemulsification with 25-gauge perfusion is a preferred method for cataract patients after vitrectomy.

OBJECTIVETo study the effect of growth hormone (GH) on the proliferation and type II collagen secretion of chondrocytes of mandibular condyle in rabbit in vitro.

METHODSFlow cytometry (FCM) and immunohistochemical technique were employed to observe the possible changes.

RESULTS(1) The exogenic GH can enhance the proliferation and synthesis of DNA of the chondrocytes of mandibular condyle in rabbit in vitro. The suitable concentration of GH is 10 microg/ml. The synthesis of DNA reaches the highest level after 12 hours, while the proliferation index (PI) hits the highest after 24 hours. (2) GH (10 microg/ml) can stimulate the secretion of type II collagen of the chondrocytes.

CONCLUSIONThe exogenic GH can enhance the proliferation, the synthesis of DNA and the secretion of type II collagen of the chondrocytes of mandibular condyle in rabbit in vitro.

Animals ; Cartilage, Articular ; cytology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chondrocytes ; drug effects ; secretion ; Collagen Type II ; drug effects ; Growth Hormone ; pharmacology ; Mandibular Condyle ; cytology ; Rabbits

[To explore the effect of Humifuse Euphorbia Herb ( HEH) on alleviating insulin resistance in type 2 diabetic KK-Ay mice. Totally 40 KK-Ay mice fed with high-fat diet were divided into four groups: the metformin group, the model group, the HEH low-dose group and the HEH high-dose group, and orally administrated with metformin hydrochloride (250 mg x kg(-1)), distilled water, humifuse euphorbia herb 1 g x kg(-1) and 2 g x kg(-1). Besides, C57BL/6J mice with ordinary feed were taken as the normal control group and orally administrated with equal distilled water. The oral administration for the five groups lasted for eight weeks. Before and after the experiment, weight, fasting glucose and insulin tolerance were determined. The morphological changes in pancreas were observed through hematoxylin-eosin (HE) staining on pancreatic tissue sections. The serum insulin, TNF-α, IL-6, adiponectin (ADPN) and leptin (LEP) were detected by ELISA. The results showed that HEH could reduce weight and fasting glucose in KK-Ay mice, alleviate hyperinsulinemia, reduce blood glucose-time AUC, increase 30-min blood glucose decline rate, relieve insulin resistance, significantly ameliorate the pathomorphological changes in pancreas in each group, decrease serum TNF-α, IL-6 and leptin levels in KK-Ay mice and rise serum ADPN level. This study proved that humifuse euphorbia herb can ameliorate the insulin resistance in KK-Ay mice, and its mechanism may be related to the effect on inflammatory factors and adipocytokines.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Euphorbia ; chemistry ; Humans ; Insulin ; metabolism ; Insulin Resistance ; Interleukin-6 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

OBJECTIVETo investigate the effects of intrauterine growth retardation (IUGR) caused by malnutrition during pregnancy on the acetylation of histone H3 and expression of histonedeacetylase1(HDAC1) in the hepar of the adult offspring and to explore the relationship between them.

METHODSMale 8-week-old offspring from maternal protein-malnutrition dams were studied. The expression of HDAC1 mRNA in the hepar was measured by fluorescent quantization RT-PCR. The levels of hepatic nuclear HDAC1 protein and acetylation of histone H3/K9 were assessed by Western blot.

RESULTSThe hepatic HDAC1 mRNA expression in IUGR rats was reduced to 54% of that of normal control rats (t=2.042, p<0.05). A decline in nuclear expression of HDAC1 protein (438 +/- 47) was also noted when compared with normal controls (1,128 +/- 110) (t=2.179, p<0.05). In contrast, the percentage of acetylated histone H3/K9 in IUGR rats (17.3 +/- 1.6%) increased significantly compared with that of normal control rats (10.5 +/- 1.2%) (t=3.597, p<0.01). The level of acetylated histone H3/K9 was negatively correlated with the HDAC1 protein concentration (r=-0.781, p<0.01).

CONCLUSIONSIncreased hepatic acetylation of histone H3 in the IUGR offspring might be caused by decreased HDAC1 expression in nuclear protein. This may contribute to the transcription change of some genes in the hepar.

Acetylation ; Animals ; Female ; Fetal Growth Retardation ; metabolism ; Histone Deacetylase 1 ; analysis ; genetics ; Histones ; metabolism ; Liver ; metabolism ; Male ; Pregnancy ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo study the validity and accuracy of differing cutoff scores of the Ages and Stages Questionnaires-Chinese (ASQ-C) for screening infants and toddlers in comparison with the gold standard, Bayley Scale of Infant Development, Second Edition (BSID II).

METHODThe 269 samples were enrolled from the normative children, aged 3 - 31 months, of the ASQ-C in Shanghai. The age-appropriate ASQ-Cs were completed by parents/caregivers and the BSIDIIwas administered by professionals. The cutoff scores of -2 standard deviation (s), -1.5 s, and -s for the ASQ-C were examined against BSID II with the cutoff scores set at -2 s as the standard of developmental delay, -s as the standard of suspected developmental delay and developmental delay respectively. Agreement between the classifications of the ASQ-C (i.e., typical, suspected, delay) was compared with the classification of the BSID II (typical, suspected, delay), sensitivity, specificity, Youden Index and area under ROC curve of ASQ-C were examined. The statistical analysis was carried out using SPSS 13.0.

RESULTWhen the cutoff score for BSID II was -2 s, the -2 s cutoff score for ASQ-C exhibited the following properties: the highest agreement of 83.64%, the sensitivity and specificity both above 80% being respectively 88.46% and 83.13%, the highest Youden Index of 0.72 and the largest area of 0.86 under ROC curve. The -1.5 s cutoff score for ASQ-C showed the following properties: 71.75% agreement, 100% sensitivity, 68.72% specificity, Youden Index = 0.69, the area under ROC curve = 0.84. The -s cutoff score for ASQ-C showed the following properties: the lowest agreement of 55.02%, 100% sensitivity, the lowest specificity of 50.21%, the lowest Youden Index of 0.50, and the smallest area of 0.75 under ROC curve. When the cutoff score for BSID II was set at -s, the -2 s for ASQ-C showed the following properties: the highest agreement of 85.87%, the lowest sensitivity of 68.57%, the highest specificity of 91.96%, Youden Index = 0.61, the smallest area = 0.77 under ROC curve. The -1.5 s for ASQ-C showed the following properties: the agreement of 80.67%, the sensitivity and specificity both above 70% being respectively 85.71% and 78.89%, the highest Youden Index of 0.65, the largest area of 0.82 under ROC. The -s cutoff score for ASQ-C showed the following properties: the lowest agreement of 68.40%, the highest sensitivity of 94.29%, the lowest specificity of 59.30%, the lowest Youden Index of 0.54, and the area under ROC curve = 0.80. When the cutoff score for BSID II was -1 to 2 s, the identifying percentages of the -2 s, -1.5 s and -s for the ASQ-C were 56.82%, 77.27% and 90.91%, respectively.

CONCLUSIONFor developmental delay identification, the -2 s cutoff score for ASQ-C produces the most robust validity and highest accuracy; for the identification of suspected developmental delay and developmental delay, the -1.5 s cutoff score for ASQ-C has the highest screening accuracy with appropriate sensitivity and specificity; for identifying the suspected developmental delay, the -s cutoff score for ASQ-C has the highest percentage of the identification. It is necessary to add 1 - 2 s to the ASQ-C's cutoff scores as the standard for screening suspected developmental delays.

Child Development ; Child, Preschool ; Humans ; Infant ; Mass Screening ; Reference Standards ; Sensitivity and Specificity ; Surveys and Questionnaires ; standards

To investigate the effects of lipopolysaccharide (LPS) on autophagy in rat hepatic stellate cells (HSC-T6) and involvement of NF-κB pathway in it.Methods 1)HSC-T6 cells were treated with LPS at the concentraction of 0, 0.01, 0.1, 1, and 10 mg/L for 0, 3, 6, 12, 18, and 24 h respectively, Microtubule-associated protein light chain Ⅱ (LC3Ⅱ) and Beclin1 levels were detected by Western blot; 2)HSC-T6 cells were randomized into groups of control group, LPS group, PDTC group, LPS+PDTC group, PMA group and LPS+PMA group, Western blot assay was used to measure the levels of LC 3Ⅱand Beclin1, immunofluoresence was used to measure NF-κB P65 intracellular distribution.The levels of hydroxyproline (Hyp) were determined by the method of colorimetry and the levels of laminin (LN) and hyaluronic acid (HA) were determined by ELISA in the culture supernatants after corresponding processing .Results The levels of LC3Ⅱand Beclin1 were significantly increased after HSC-T6 cells were treated with LPS at the concentraction of 0.1 mg/L for 6 h and the levels of Hyp, LN, and HA in the culture supernatants increased remarkably as well (P<0.05).PDTC pretreatment increased the levels of LC3Ⅱand Beclin1 in the LPS-treated HSC-T6 cells while decreased the levels of Hyp , LN, and HA significantly (P<0.05).PMA pretreatmen decreased the levels of LC 3Ⅱand Beclin1 in the LPS-treated HSC-T6 cells while increased the levels of Hyp, LN, and HA (P<0.05).Conclusions LPS can promote autophagy and activation of NF -κB pathway in HSC-T6 cells.Activation of NF-κB pathway may inhibit the LPS-induced autophagy of HSC-T6 cells.

Objective: This study aimed to assess the technical feasibility, efficacy, and safety of the safe triangular working zone (STWZ) approach applied in percutaneous vertebroplasty (PV) for spinal metastases involving the posterior part of the vertebral body. Materials and Methods: We prospectively enrolled 87 patients who underwent PV for spinal metastasis involving the posterior part of the vertebral body, with or without the STWZ approach, from January 2019 to April 2022. Forty-nine patients (27 females and 22 males; mean age ± standard deviation [SD], 57.2 ± 11.6 years; age range, 31–76 years) were included in group A (with STWZ approach), accounting for 54 vertebrae. Thirty-eight patients (18 females and 20 males; 59.1 ± 10.9 years; 29–81 years) were included in group B (without STWZ approach), accounting for 57 vertebrae. Patient demographics, procedure-related variables, and pain relief as assessed using the visual analog scale (VAS) were collected at different time points. Tumor recurrence in the vertebrae after PV was analyzed using Kaplan–Meier curves. Results: The STWZ approach was successful from T1 to L5 without severe complications. Cement filling was satisfactory in 47/54 (87.0%) and 25/57 (43.9%) vertebrae in groups A and B, respectively (v< 0.001). Cement leakage was not significantly different between groups A and B (p= 1.000). Mean VAS score ± SD before and 1 week and 1, 3, 6, 9, and 12 months after PV were 7.6 ± 1.8, 4.2 ± 2.0, 2.7 ± 1.9, 1.9 ± 1.5, 1.7 ± 1.4, 1.7 ± 1.1, and 1.6 ± 1.3, respectively, in group A and 7.2 ± 1.7, 4.0 ± 1.3, 3.4 ± 1.6, 2.4 ± 1.2, 1.8 ± 1.0, 1.4 ± 0.5, and 1.7 ± 0.9, respectively, in group B. Kaplan–Meier analysis showed a lower tumor recurrence rate in group A than in group B (p = 0.001). Conclusion The STWZ approach may represent a new, safe, alternative/auxiliary approach to target the posterior part of the vertebral body in the PV for spinal metastases.

Nephronophthisis (NPHP) is a group of autosomal recessive tubulointerstitial cystic kidney disorders. This article reports a case of NPHP type 12 caused by TTC21B mutations. The girl had an insidious onset, with moderate proteinuria, renal dysfunction, stage 2 hypertension, situs inversus, and short phalanges when she visited the hospital for the first time at the age of 3 years and 6 months. The renal lesions progressed to end-stage renal disease (ESRD) before she was 4 years old. Urine protein electrophoresis showed glomerular proteinuria. There were significant increases in urinary β2-microglobulin and α1-microglobulin. Gene detection revealed two compound heterozygous mutations, c.1552T>C (p.C518R) and c.752T>G (p.M251R), in the TTC21B gene, which came from her father and mother respectively. The c.752T>G mutation was a novel mutation. It is concluded that besides typical tubular changes of NPHP, marked glomerular damage is also observed in patients with TTC21B gene mutations.
Child, Preschool ; Female ; Genotype ; Humans ; Kidney ; Kidney Diseases, Cystic ; Kidney Failure, Chronic ; Microtubule-Associated Proteins ; genetics ; Mutation ; Nephrosis ; genetics

BACKGROUNDThe aim of this study was to assess the performance of apparent diffusion coefficient (ADC) measurement obtained with diffusion-weighted magnetic resonance imaging (DW-MRI) to distinguish renal cell carcinomas (RCCs) from small benign solid renal tumors (≤ 4 cm).

METHODSIn this cross-sectional study, 49 consecutive patients with histopathologically confirmed small solid renal tumors, and seven healthy volunteers were imaged using nonenhanced MRI and DW-MRI. The ADC map was calculated using the b values of 0, 50, 400, and 600 s/mm 2 and values compared via the Kruskal-Wallis and Mann-Whitney tests. The utility of ADC for differentiating RCCs and benign lesions was assessed using a receiver operating characteristic curve. Multiple nonenhanced MRI features were analyzed by Logistic regression.

RESULTSThe tumors consisted of 33 cases of clear-cell RCCs (ccRCCs) and 16 cases of benign tumors, including 14 cases of minimal fat angiomyolipomas and 2 cases of oncocytomas. The ADCs showed significant differences among benign tumors ([0.90 ± 0.52] × 10-3 mm 2 /s), ccRCCs ([1.53 ± 0.31] × 10-3 mm 2 /s) and the normal renal parenchyma ([2.22 ± 0.12] × 10-3 mm 2 /s) (P < 0.001). Moreover, there was statistically significant difference between high and low-grade ccRCCs (P = 0.004). Using a cut-off ADC of 1.36 × 10-3 mm 2 /s, DW-MRI resulted in an area under the curve (AUC), sensitivity, and specificity equal to 0.839, 75.8%, and 87.5%, respectively. Nonenhanced MRI alone and the combination of imaging methods led to an AUC, sensitivity and specificity equal to 0.919, 93.9%, and 81.2%, 0.998, 97%, and 100%, respectively. The Logistic regression showed that the location of the center of the tumor (inside the contour of the kidney) and appearance of stiff blood vessel were significantly helpful for diagnosing ccRCCs.

CONCLUSIONSDW-MRI has potential in distinguishing ccRCCs from benign lesions in human small solid renal tumors (≤ 4 cm), and in increasing the accuracy for diagnosing ccRCCs when combined with nonenhanced MRI.

Adult ; Aged ; Carcinoma, Renal Cell ; diagnosis ; Cross-Sectional Studies ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Kidney Neoplasms ; diagnosis ; Male ; Middle Aged ; Young Adult