OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

According to the correlative analyses on Chinese medicine essence, dosage forms and quality control level, it expounds the precise concept of Chinese medicine, and its quality advantages and characteristics in this paper, furthermore discusses how to achieve the ideal drugs and Chinese medicine quality precision in expectation. Base on the Chinese medicine essence, using the concept of nature medicine and its drug system to construct Chinese medicine effective material basis and its drugs, with the correlative analyses of whole view and reductionism, the problems of uncertainty quality of original natural medicinal resources and preparations may well be solved, and further with the macroscopic to microcosmic construction of drug system, the precision in expectations of Chinese medicine quality and higher production lever may well be achieved.
Drugs, Chinese Herbal ; chemistry ; pharmacology ; standards ; Humans ; Medicine, Chinese Traditional ; Plants, Medicinal ; chemistry ; Precision Medicine ; Quality Control

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

HSP90, which is the biomarker of cell stress and endogenous protective protein, functions as a molecular chaperone. Many client proteins of HSP90, including EGFR, Met, Raf-1, IKK and p53, play important roles in the occurrence and development of tumor. Binding of HSP90 inhibitors triggers the deactivation of HSP90, resulting in client protein degradation, and hence inhibits the tumor growth by blocking multiple targets involved in signaling of tumor proliferation. This review summarizes recent development of small molecule inhibitors bound to N-terminal of HSP90.

Objective To explore the influence of intestinal trefoil factor (ITF) on interleukin-6(IL-6) in neonatal rat with necrotizing enterocolitis(NEC) ,and to discuss the protective machanism of ITF on NEC.Methods Thirty-two neonatal rats were divided randomly into four groups,group A as control group,group B as NEC group,group C as NEC+NS 0.2 mL group,group D as NEC+ITF 0.2 mg group.NEC model of neonatal rats were established.On the 4th day,all the subjects were put to death.Intestinal tissue within the boundary of ileum and cecum was obtained to observe histological changes.Other intestinal tissue was treated into homogenate.After the homogenate was centrifuged,supernates were used to test the density of IL-6.Results The density of IL-6 significantly decreased in group A,D than those in group B and C (Pa0.05).The pathological lesions indicated that intestinal tissue necrosis was severe in group B and C,which was graded as 3 points,but obviously lessen in group D,which was graded as 1 point,with ITF interfering.Conclusions Intestinal inflammation is ameliorated after ITF are injected hypodermically or intraperitoneally.ITF may provide a brand-new way for the therapy of NEC in neonatal rats.

In this paper, the process performance indexes (PPIs) P(p) and P(pk) were introduced and applied to evaluate the process capability and quality consistence of Chinese medicine products. The historical quality analysis data of Qingkailing injection were collected and taken as the research object. The confidence intervals of P(p) and P(pk) were estimated based on the Bootstrap sampling methods. Results showed that the value and width of the confidence interval P(p) of were smaller that of P(pk) suggesting that the P(pk) index was more sensitive than P(p) in process capability analysis. Within the production period concerned, the P(pk) values estimated from different quality indexes of Qingkailing injection, such as baicalin, cholic acid, geniposide and total nitrogen, were 1.122, 2.055, 1.564 and 0.891, respectively. It could be found that the cholic acid had the highest process capability, followed by the geniposide and baicalin. The total nitrogen had the lowest process capability, indicating that it is necessary to reinforce the quality management of total nitrogen related manufacturing processes. The case studies demonstrate the effectiveness and feasibility of PPIs, which are convenient to be used in production practice of Chinese medicine.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; Injections ; Quality Control

OBJECTIVETo observe the intervention effects of electroacupuncture at "Yishu" (EX-B 3) on rats with type-2 diabetes mellitus (T2DM), so as to provide experiment references for acupuncture to treat T2DM.

METHODSAmong seventy male Wistar clean-grade rats, 8 rats were randomly selected into a control group; the rest rats were made T2DM model. Fifty-two rats which were successfully made T2DM model, according to randomized block method, were divided into a model group (10 rats), a medication group (10 rats), an electroacupuncture at "Shenshu" (BL 23) group (11 rats), an electroacupuncture at "Pishu" (BL 20) group (10 rats) and an electroacupuncture at "Yishu" (EX-B 3) group (11 rats). Seven days after successful establishment of model, the rats in the model group were fixed in the self-made rat bag without receiving any treatment; the rats in the medication group, according to body mass (10 mL/kg), were treated with intragastric administration of glimepiride; the rats in all the electroacupuncture groups were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Yishu" (EX-B 3), respectively. The continuous wave was selected with a frequency of 15 Hz and a current intensity of 4 to 6 mA. The treatment was given 20 min per treatment, once a day, 5 treatments per week for continuous 4 weeks. Before the establishment of model and continuous 4 weeks after the intervention, blood samples were collected from rats' caudal vein, and fasting blood glucose (FBG) was measured with FBG device each week. After the last intervention, the rats were killed and hypothalamus, pituitary and adrenal gland were collected. The colorimetric method was applied to measure the contents of triglyceride (TG), cholesterol (TC), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C); radioimmunoassay was used to test the contents of glycated serum protein (GSP), fasting insulin (FINS), corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortin (CORT).

RESULTSFour weeks after the intervention, except that the rat's body mass in the normal group continued to increase, body mass in the model group, medication group and each electroacupuncture group were significantly reduced (P<0.01, P<0.05). Compared with the model group, the FBG in the electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu" (EX-B 3) group were obviously reduced (P<0.05, P<0.01); FBG in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05). The contents of TG, HDL-C and LDL-C in the electroacupuncture at "Yishu" (EX-B 3) group were reduced (P<0.05, P<0.01), the content of TG was significantly lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05), the content of LDL-C was significantly lower than that in electroacupuncture at "Shenshu" (BL 23) group (P<0.05). Insulin sensitivity index (ISI) in the medication group, electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu (EX-B 3)" group were evidently increased (P<0.05, P<0.01); ISI in the medication group was lower than that in the electroacupuncture at "Yishu" (EX-B 3) group (P<0.05). The content of CRH in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (P<0.05, P<0.01); the content of CORT in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Pishu" (BL 20) group (both P<0.05).

CONCLUSIONElectroacupuncture at "Yishu" (EX-B 3) could reduce the level of CORT to improve the insulin resistance in rats with T2DM, improve insulin sensitivity index, regulate blood lipid metabolism and relieve the hyperactivity of the HPA axis.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; therapy ; Electroacupuncture ; Hormones ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo detect the expressions of receptor tyrosine kinases (RTKs) mRNA and protein and to explore potentially promising tumor markers and conceivable drug target in bladder cancer.

METHODSThe expressions of RTKs mRNA and protein in tissue from invasive urothelial carcinoma of the bladder were examined by real-time quantitative PCR array and cytokine antibody array, with normal bladder tissue as control. The Results were analyzed using bioinformatic approaches.

RESULTSThe expressions of TGFA, STAB1, SERPINE1, ANGPT2, SPINK5, ANGPTL1, PROK1, MDK, CXCL9, GRN, RUNX1, VEGFA, and TGFB1 were obviously upregulated in bladder cancer tissue, while those of EDIL3, PTN, CCL2, PDGFD, FGF13, KITLG, FGF2, SERPINF1, and TNF were downregulated. ALK, Btk, EphB2, ErbB4, PDGFR-α, ROS, Tie-2, Tyk2, and VEGFR3 were over-expressed in bladder cancer, while FRK, Fyn, IGF-IR, Insulin R, Itk, JAK1, JAK3, and LCK were low-expressed.

CONCLUSIONVascular endothelial growth factor/platelet-derived growth factor-targeted therapies may play an active role in treating carcinoma of bladder.

Carcinoma, Transitional Cell ; metabolism ; Humans ; RNA, Messenger ; genetics ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Urinary Bladder Neoplasms ; metabolism

OBJECTIVETo explore the values of adrenocorticotropic hormone receptor (ACTH-R) determination and ultrastructural observation of tumor cells in the subtyping of adrenocortical neoplasms (ANs).

METHODSThe expression of ACTH-R in 87 AN tissues were determined with Polymer immunohistochemical staining, with 10 normal adrenal tissues as the controls. The ultrastructure of the tumor cells was observed using electron microscopy.

RESULTSThe positive expression rate of ACTH-R was (80.1 +/- 8.2)%, (53.2 +/- 10.3)%, (63.2 +/- 10.1)%, (83.3 +/- 6.5)%, and (70.1 +/- 7.3)% in the sub-CPA group, CPA group, APA group, NFA group, and NC group, respectively. ACTH-R expression was significantly higher in NFA and sub-CPA groups than in NC group (P = 0.001, P = 0.000), APA group (P = 0.000, P = 0.000), and CPA group (P = 0.000, P = 0.000), and was also significantly different between NC group and APA group (P = 0.039) and between APA group and CPA group (P = 0.037). However, no significant difference was found between NFA group and sub-CPA group (P = 0.325). As shown by the electron microscopy, ANs had some partially similar microscopic features, while different AN subtypes showed differences in the type and amount of secretory granules.

CONCLUSIONACTH-R determination and ultrastructural observation of tumor cells may be helpful for subtyping ANs.

Adrenal Cortex Neoplasms ; diagnosis ; metabolism ; ultrastructure ; Adrenal Glands ; metabolism ; ultrastructure ; Adult ; Female ; Humans ; Male ; Middle Aged ; Receptors, Corticotropin ; metabolism

OBJECTIVETo summarize the efficacy of open stone surgery for renal and ureteral calculi, and to investigate the current indications for upper urinary tract stones treated by open stone surgery.

METHODSTo analyze retrospectively the cases with upper urinary tract calculi treated by open stone surgery from January 1995 to December 2004.

RESULTSIn all, there were 667 patients treated for renal and ureteral calculi by open stone surgery, at the same time 9000 cases with upper urinary tract calculi also treated in our centre with open surgery rate 7.4%. Indications for open surgery were complex stone burden of 297 cases, nephrectomy for non-functioning kidneys of 137 cases, with anatomical abnormalities of 134 cases, ESWL or endoscopy failure of 57 cases, with co-morbid medical diseases of 30 cases, with renal carcinoma of 5 cases, with xanthogranulomatous pyelonephritis of 3 cases, with kidney injury of 2 cases, and with other ipsilateral operation of 2 cases. Of 145 cases for nephrectomy, 16 patients and 6 patients suffered from various intraoperative and postoperative complications respectively;of 522 patients treated by open stone surgery, 7 patients and 34 patients were suffered from various intraoperative and postoperative complications respectively; 57 patients with multiple renal calculi had residual calculi with overall stone-free rate 89.1%. And no case died in perioperative period.

CONCLUSIONSOpen surgery maintains its important role for treatment of renal and ureteral calculi because of its safety and efficacy. Nephrectomy for nonfunctioning kidney with renal and/or ureteral calculi, huge and complex renal calculi, anatomical abnormalities, minimal invasive therapies failure and some selective patients with co-morbid medical diseases are suitable for open stone surgery with high success and low morbidity rate.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Kidney Calculi ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Ureteral Calculi ; surgery ; Young Adult