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.

OBJECTIVERNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.

METHODSThe pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.

RESULTSSEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).

CONCLUSIONResults indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.

Animals ; Cleft Palate ; Gene Silencing ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; Oxidoreductases Acting on CH-CH Group Donors ; Palate ; growth & development ; RNA, Messenger

OBJECTIVEThe purpose of this study was to study the masticatory muscles affected by distraction osteogenesis of the mandible.

METHODSThe distraction osteogenesis (DO) was applied to distract the left mandible of 6 mongrel dogs that were divided into three experimental groups. After different distraction phase and consolidation phase, the masseter and the digastric muscle were taken out. The specimens were stained using hematoxylin/eosin and enzyme histochemistry. Afterwards, the specimens were observed with a light microscope to study the morphologic changes of the muscles. The contents of enzyme in the different groups were measured by VIDAS.

RESULTSThe masseter showed consequently atrophy, but the digastric muscle showed a progress of histomorphologic reconstruction, including atrophy and hypertrophy. The changes of the contents of enzyme and histomorphology were identical in the masticatory muscles.

CONCLUSIONThe digastric muscle parallel to the vector of mandibular distraction adapts the distraction by the way of atrophy, regeneration and hypertrophy. And the contents of enzyme appear to decrease at the beginning, increase afterwards, and return to the normal level finally. But the masseter perpendicular to the vector of mandibular distraction shows consequent atrophy, and the contents of enzyme consequently decrease, which means the metabolism decrease.

Adenosine Triphosphatases ; analysis ; Animals ; Dogs ; Female ; L-Lactate Dehydrogenase ; analysis ; Male ; Mandible ; surgery ; Masticatory Muscles ; anatomy & histology ; enzymology ; Osteogenesis, Distraction ; Random Allocation

The in-vitro fertilization and embryo transfer technique has been widely applied in human insemination. The rate of successful insemination is gradual y rising, and the in-vitro fertilization directly determine the insemination outcome.
OBJECTIVE:To evaluate the difference between the two common using insemination methods, microdrop and open, in in-vitro fertilization and embryo development.
METHODS:A randomized study was conducted to compare microdrop and open insemination methods among non-male factor patients undergoing in-vitro fertilization and embryo transfer. A total of 1 175 cases were enrol ed in the research. There were 573 cases in the microdrop group, and 602 cases in open insemination group. The fertilization rate and embryo development in the two groups were compared.
RESULTS AND CONCLUSION:The fertilization failure rate [total fertilization failure rate+low fertilization rate (<25%oocytes fertilized)] in the microdrop insemination group was higher than in the open insemination group (11.9%, 3.3%, P<0.001), while the good quality embryo rate and pregnancy rate did not differ significantly between the two groups (al P>0.05). The open insemination method is a simple insemination method with a lower fertilization failure rate. As the fertilization is a highly complicated process involving many extrinsic and intrinsic factors, further study is needed to confirm the effects of the two insemination methods on in-vitro fertilization outcome.

This article is aimed to provide a method for the simultaneous analysis of five heavy metals, including Cu, As, Pb, Cd, and Hg, in Chuanxiong Rhizoma and Gastrodiae Rhizoma through ICP-MS. Five heavy metals were determined by an inductively coupled plasma-mass spectrometry method after microwave-assisted digestion. The linear correlation coefficients were all better than 0.999. The lowest limits of quantification for all target elements were 0.003 0.134 ug·L-1, while the recovery values ranged from 80.04% 118.34%. This method was accurate, convenient, rapid, and highly sensitive, and can be applied to determine five heavy metals, including Cu, As, Pb, Cd, and Hg in Chuanxiong Rhizoma and Gastrodiae Rhizoma.

BACKGROUND:Recombinant bovine basic fibroblast growth factor is a manifold effect cytokine which can promote angiogenesis, wound healing, tissue repair and bone regeneration. Recombinant bovine basic fibroblast growth factor with good histocompatibility is easy to operate and has been widely used in oral and maxil ary surgery.
OBJECTIVE:To evaluate the effect of recombinant bovine basic fibroblast growth factor against dry socket syndrome after tooth extraction.
METHODS:A total of 160 patients who had been extracted mandibular third molar were selected and randomly divided into two groups. In the experimental group, recombinant bovine basic fibroblast growth factor was put into the sockets after mandibular third molars were extracted, while in the control group, we let the wounds to be healed natural y without any materials. The incidence of dry socket syndrome was observed and compared between two groups at 3 days, 5 days and 1 week after tooth extraction.
RESULTS AND CONCLUSION:One patient had dry socket after operation in the experimental group, and the incidence was 1.25%. In the control group, 10 patients suffered from dry socket, and the incidence was 12.5%. There was a significant difference in the incidence of dry socket between the two groups (P<0.01). There was visible granulation tissue within the tooth socket after tooth extraction in the experimental group, and extraction sockets narrowed and were fil ed with granulation tissues, which was 1-2 days earlier than the control group. No al ergies, tissue hyperplasia and other local and systemic reactions occurred in patients receiving implantation of recombinant bovine basic fibroblast growth factor gel. These findings indicate that local implantation of recombinant bovine basic fibroblast growth factor gel after mandibular tooth extractions can speed up the healing of dental extraction wounds.

This study was aimed to establish the determination method of nineorganic solvent residues of macrop-orous resin. Headspace capillary gas chromatography (GC) was used in the content determination of hexane, benzene, methyl-benzene, o-xylene, m-xylene, p-xylene, styrene, diethyl-benzene and divinyl-benzene of macroporousresin residues in Ginkgo leaf tablets. The results showed that ninemacroporous resin residues in Ginkgo leaf tablets were completely separated. The calibration curves were in good linearity (r=0.9992-0.9998). The average recoveries were 80%-110%. It was concluded that themethod was accurate, quick, and sensitive, which can be used to determine the macroporous resin residues in Ginkgo leaf tablets.

OBJECTIVETo investigate the effect of 7- dehydrocholesterol reductas (Dhcr-7) gene silencing on the palatal development by sonic hedgehog (Shh)-bone morphogenetic protein2(BMP-2) signal pathway in vitro.

METHODSA total of 60 pairs of palatal shelves fromgestation day (GD) 13.5 mouse embryos were divided into three groups (A, B, C) of 20 randomly. In group A (control), palatal shelves were cultured with medium containing no cholesterol.In group B (Dhcr-7-siRNA), palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus. After 48h, the culture medium of groups A and B were changed with medium without cholesterol. In group C (cholesterol), palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus. After 48h, the culture medium of group C was changed with medium containing 600 mg/L cholesterol. After 72h again, tissues dyeing and scanning electron microscope (SEM) technique were used to observe morphological changes of palates. Both RT-PCR and Western blottingtechniques were used to measure mRNA and protein expressions for Dhcr-7, Shh, and BMP-2, respectively.

RESULTSThe tissues dyeing and SEM showedthat the palates fusedin groups A and Candthe palates did not fuse in group B eventually. The expression of both mRNA and proteins for Shh and BMP-2 in group B wasdecreased with the Dhcr-7 reduction. In group B, the mRNA and protein expression of Shh was separately 0.063±0.018 and 0.092±0.065;the mRNA and protein expression quantity of BMP- 2 was separately 0.054±0.018 and 0.049±0.021. In group A, the mRNA and protein expression of Shh was separately 0.667±0.093 and 0.639±0.078;the mRNA and protein expression of BMP-2 was separately 0.591 ± 0.043 and 0.569 ± 0.081. The difference of Shh and BMP- 2 mRNA and protein expression between A and B group were statistically significant separately (P < 0.05). The expression of both mRNA and protein for Dhcr-7 (0.074±0.034 and 0.075±0.028) did not changebasicallyin group C, compared with the Dhcr- 7expression of mRNA and protein (0.083±0.045; 0.067±0.065) in group B, the difference wasnot statistically significant(P > 0.05). In group C, the mRNA and protein expressionof Shh (0.649±0.085 and 0.608±0.092) and BMP-2 (0.578±0.062 and 0.548±0.065) were significantly increased. The difference of Shh and BMP-2 mRNA and protein expression between B and C group were statistically significant separately (P < 0.05).

CONCLUSIONSDhcr-7 could influence the expression of Shh and BMP-2. Dhcr-7 reductase regulated the palatal development by the Shh-BMP-2 signal pathway.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cholesterol ; Culture Media, Conditioned ; chemistry ; pharmacology ; Hedgehog Proteins ; genetics ; metabolism ; Mice ; Oxidoreductases Acting on CH-CH Group Donors ; metabolism ; Palate ; growth & development ; metabolism ; RNA, Messenger ; analysis ; metabolism ; RNA, Small Interfering ; metabolism ; Random Allocation ; Signal Transduction

OBJECTIVETo evaluate the osteogenesis of alveolar bone graft (ABG) in patients with alveolar cleft by cone beam CT (CBCT).

METHODSABG surgery was performed in 20 patients with unilateral complete alveolar cleft. The patients were followed up for 3 and 6 months after surgery and the osteogenesis of the bone graft was evaluated by CBCT. The bone density and the height of labial and palatal bone graft area were analyzed.

RESULTSThere was no significant difference in the bone density between 3 months [(403.79 ± 64.70) HU] and 6 months[(411.45 ± 42.62) HU ] (P = 0.329).However, there was significant difference in bone height in the labial and palatal side between 3 months and 6 months (labial P = 0.020, palatal P = 0.008).

CONCLUSIONSThe osteogenesis was the best 3 months after bone graft. The following treatment can start in this stage.

Bone Transplantation ; Cleft Palate ; Cone-Beam Computed Tomography ; Humans ; Osteogenesis

Objective To investigate the expression level of transforming growth factorβ1 (TGF-β1) and interleukin 17 (IL-17) in patients with allergic conjunctivitis. Methods TGF-β1 and IL-17 expression levels in allergic conjunctivitis patients hospitalized in the author's hospital and a healthy population were detected to explore the expression level in all types of patients with allergic conjunctivitis. Results TGF-β1 and IL-17 levels were significantly higher in patients with allergic conjunctivitis than in healthy population (P<0.01). Among the four clinical subtypes of allergic conjunctivi-tis, vernal keratoconjunctivitis had the highest expression level of TGF-β1 and IL-17 in spring, followed by perennial allergic conjunctivitis and seasonal allergic conjunctivitis, and atopic keratoconjunctivitis had the lowest expression level. However, the differences among the four subtypes were not significant (P>0.05). Conclusion Patients with allergic conjunctivitis have higher TGF-β1 and IL-17 expression levels than normal healthy people. TGF-β1 and IL-17 ex-pression level vary in different subtypes of allergic conjunctivitis, vernal keratoconjunctivitis has higher expression level than other types in spring.