The study is aimed to explore a rapid method to extract DNA from fried Chinese medicinal products. The alkaline lysis buffer was made of sodium hydroxide, 1% PVP and 1% TritonX-100 and Tris-HCl solution was neutralized, through heat cracking and neutralization two step to extract DNA from processed and prepared products of traditional Chinese medicine. Then universal primes were used to amplify PCR products for fired Chinese medicinal materials. The results indicated the optimized alkaline lysis method for extracting DNA is quick and easy. Extracting of the different processed Sophora japonica of DNA concentration was (420.61 ± 123.91) g x L(-1). Using 5% Chelex-100 resin purification can improve the DNA concentration. Our results showed that the optimized alkaline lysis method is suitable for Chinese medicinal materials for quickly DNA extraction.
Alkalies ; chemistry ; Chemical Fractionation ; methods ; DNA, Plant ; genetics ; isolation & purification ; Hydrolysis ; Plants, Medicinal ; chemistry ; classification ; genetics ; Polymerase Chain Reaction ; Sophora ; chemistry ; classification ; genetics

Fosmidomycin (100 micromol x L(-1)) which is the effective inhibitor of DXR, key enzyme in terpenoid MEP pathway, was used to treat with hairy roots of Salvia miltiorrhiza. The treated roots were harvested at 2, 4, 6, 8, 10, 16 and 21 d, mRNA level of SmDXR and tanshinone content in treated and negative control groups were detected. Results found that, after treated with fosmidomycin, color of S. miltiorrhiza hairy roots grew pale gradually comparing with controls; mRNA level of SmDXR in hairy roots varied as a shape of parabolic and the highest value achieved at the sixth day after treatment, then it decreased gradually; Content of four kinds of tanshinones were detected. Among of the four kinds of tanshinones, Tanshinone I content changed relatively little, while content of dihydrotanshinone I, cryptotanshinone and tanshinone II (A) decreased gradually in 21 days. The content of total tanshinones in NC groups was 5, 63 times more than FOS-treated roots in the 21th day. The previous results showed that SmDXR played an important role in the accumulation of tanshinone content in MEP pathway. Once the mRNA level of SmDXR was suppressed, the accumulation of secondary metabolites will be significantly affected.
Aldose-Ketose Isomerases ; genetics ; Diterpenes, Abietane ; metabolism ; Fosfomycin ; analogs & derivatives ; pharmacology ; Gene Expression Regulation, Plant ; drug effects ; Plant Roots ; drug effects ; growth & development ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; growth & development ; metabolism ; Time Factors

AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.

Objective To compare the diagnosis results of endemic skeletal fluorosis from clinical and X-rays examinations, in order to provide the foundation for revising clinical diagnostic standard of endemic skeletal fluorosis. Methods The 675 inhabitants aged 16 to 60 years old were retrospectively chosen as subjects in 15 villages drinking un-improved water, where they lived for 10 years or more. Drinking water fluoride were rated as 0.5,1.0, 1.5,2.0,2.2,2.4,3.0,3.5,4.0,6.0,7.0 mg/L levels in Qianan and Nongan County of Jilin Province. The clinical and X-rays results of endemic skeletal fluorosis were analyzed and compared at different drinking water fluoride levels. Results The clinically detectable rates of endemic skeletal fluorosis(21.43%,22.45% ,21.28%, 19.05%, 38.89%) were higher than that of X-rays(0,2.04%,0,4.76%, 12.96%, X2=7.96,9.49,11.19,4.08,9.45, P<0.05) when fluoride content of drinking water was 2.0,2.2,2.4,3.0,4.0 mg/L. X-rays detective rates were 0 at water fluorides levels of 2.0,2.4 mg/L and still low at water fluoride levels of 3.0,4.0 mg,/L. The difference of detective rates of endemic skeletal fluorosis between the clinical (1.00%,4.44%, 7.23%, 18.00%, 54.39%, 49.18%) and X-rays (0,2.22%, 3.61%, 8.00%, 36.84%, 52.46%) were not statistically significant at water fluorides levels of 0.5,1.0,1.5,3.5,6.0,7.0 mg/L(X2=1.00,0.17,0.47,2.21,3.54,0.13, P>0.05). Conclusions The detectable rates of skeletal fluorosis increase with the increased concentration of water fluoride, which is more reliable for clinical examination than for X-rays method.

Objective To observe the long term effects of arthroscopic knee debridement and reconstructing operation for treating osteoarthritis in patients with Kaschin-Beck disease. Methods Thirty-one cases of patients with Kaschin-Beck disease were followed for 6 years after operation of articular clearing by arthroscope. Index of pain, symptoms of self-evaluation, range of motion, walking distance, standing test by affected leg when bending at 30° or 60° were recorded and compared with the preoperative results. Results Twenty-four cases were followed up for 6 years. Six years after operation the pain index(3.38 ± 2.87) was dramatically decreased compared to that before operation (6.88 ± 1.45, t = 5.30, P ＜ 0.05). Patients symptoms markedly improved by subjective self-evaluation was 70.83% (17/24), the effective rate was 100% (24/24). The number of cases that could stand up when leg bending at 30° or 60° were 21,18 cases, respectively, compared with that of preoperative of 14, 11 cases, respectively, the difference was statistically significant(x2 = 5.17,4.27, all P ＜ 0.05). Six years after operation the walking distance(3 cases ＜ 1 km, 11 cases 1 - 5 km and 10 cases ＞ 5 km) were greatly improved compared to the results before operation (12 cases ＜ 1 km, 9 cases 1 - 5 km and 3 cases ＞ 5 km, U = 2.88, P ＜0.05). Six years after operation the knee activity[(132.25 ± 14.52)°] remained at the same level, compared with that of preoperative [(131 .58 ± 14.68) °], the difference was not statistically significant (t = 0.16, P ＞ 0.05) .Conclusions The method of arthroscopic joint debridement to cure Kaschin-Beck disease knee osteoarthritis can significantly reduce pain, improve function and walking distance, with more stable long-term satisfactory outcome.

OBJECTIVE: To explore the application value of postmortem multi-slice spiral computed tomography (MSCT) by observing and analyzing the injury features in the traffic accident victims. METHODS: Ten traffic accident victims were scanned with whole body MSCT. The systemic autopsy was subsequently performed to compare with the results of MSCT. The advantages and disadvantages of autopsy and MSCT for obtaining the information of traffic accident injuries were then analyzed. RESULTS: MSCT could reveal 3D shape of fractures clearly and detect air accumulation in different positions of the body, which showed the obvious advantages compared with autopsy. However, the resolution of MSCT was limited compared to the detection of organ and soft tissue injuries. CONCLUSION A combination of MSCT and autopsy is the best way for determining the manner and the cause of death in traffic fatality victims.
Accidents, Traffic ; Autopsy ; Fractures, Bone ; Humans ; Soft Tissue Injuries ; Tomography, Spiral Computed

High resolution melting (HRM), an important technology for genotyping and mutation scanning, has broad prospects in the authentification of traditional Chinese medicine. This paper selected universal trnH-psbA primers and used HRM to establish a new methods for identification of Akebia herbs. PCR was conduct at the annealing temperature of 58 degrees C and 35 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further analyzed. The results showed the Tm values of Caulis Akebiae was (81.84 ± 0.16), (85.28 ± 0.16) degrees C and Caulis Clematidis Armandii was (83.22 ± 0.19) degrees C and Caulis Aristolochiae manshuriensis was (81.67 ± 0.14) degrees C, (84.24 ± 0.10) degrees C with 5-125 mg - L-' DNA template, 0.4 μmol x L(-1) primer, 2.0 mmol x L(-1) Mg2+. This method can achieve the authentification of Akebia herbs and is simple, fast, high-throughput, visual.
Chemistry Techniques, Analytical ; methods ; DNA, Plant ; chemistry ; genetics ; Genotype ; Magnoliopsida ; chemistry ; classification ; genetics ; Phylogeny ; Transition Temperature

Objective To monitor and compare the breathing mechanics on PC,VC and PRVC during pneumoperitoneum,and to discuss the significance of the clinic use of PRVC.Method Ninety laparoscopic cholecystectomy patients were equally divided into 3 groups (PC,VC,PRVC).Levels of PES,PAWM,PAP,PaCO2,ETCO2,TV MAP and HR were detected before pneumoperitoneum,and at 5,10,15 and 20 minutes postpneumoperitoneum.Results Pneumoperitoneum made three respiratory patterns with different levels of PAWM,PAP,and PES.PES post-pneumoperitoneum in the VC model was obviously higher than that in the PC and PRVC group.At 10 min post-pneumoperitoneum,levels of PaCO2 and ETCO2 increased obviously in the PC and VC group(P ＜ 0.05).Levels of PaCO2 and ETCO2 were increased in the PC group,but TV level post-pneumoperitoneum was significantly lower than that in the other two groups (P ＜ 0.05).Level of PaCO2 and ETCO2 were increased in the PC and VC group post-pneumoperitoneum,along with increases of MAP and HR (P ＜ 0.05).Levels of MAP and HR in the PRVC group post-pneumoperitoneum were significantly lower than those in the PC and VC group (P ＜ 0.05).Conclusion PRVC mode can effectively reduce the increases of pneumoperitoneum-induced PAWM,PAP,PES,without the unusual increase of PaCO2 and ETCO2 during surgeries,contributing to the stability of vital signs of perioperative patients.

Objective To evaluate the clinical results of arthroscopic repair of combined Bankart and superior labrum anterior to posterior (SLAP)lesions in patients with recurrent shoulder dislocations.Methods Between May 2011 and January 2015,we reviewed 15 cases with combined Bankart and SLAP lesions with recurrent shoulder dislocations who underwent arthroscopic repair.Their average age during surgery was 24.2 years (ranging from 16 to 38 years).During the operation,we began by repairing the unstable SLAP lesion with absorbable suture anchors before we repaired Bankart lesion from the inferior to superior.Fifteen patients in the control group had isolated Bankart lesions without SLAP lesions and underwent arthroscopic repair.Their mean age was 24.6 years (ranging from 18 to 35 years).The preoperative and postoperative results were analyzed by Visual Analogue Scale (VAS)for pain,the range of motion,American Shoulder and Elbow Surgeon (ASES)and Rowe Shoulder Scores Systems.We compared the results with the isolated Bankart lesion. Results For patients who underwent arthroscopic repair of combined Bankart and SLAP lesions,the mean postoperative follow-up period was 15 months (ranging from 13 to 28 months),vs 22 months (ranging from 21 to 34 months)in the control group.VAS for pain was decreased from preoperative 4.9 to postoperative 1.9 (P <0.05).Mean ASES and Rowe Shoulder Scores were improved from preoperative 56.4 and 33.7 to postoperative 91.8 and 94.1,respectively (P <0.05). Recurrent dislocation was not observed until the last follow-up and anterior instability in two groups was not noted during the physical examination.Compared with the isolated Bankart lesion group,no statistical significance was found in the ASES score,Rowe Shoulder Score,or VAS for pain (P >0.05 ).Anterior flexion,abduction,and internal rotation of the affected shoulder during the last follow-up were normal after surgery.Conclusion For recurrent dislocation of the shoulder with combined Bankart and SLAP lesion,arthroscopic repair using absorbable suture anchors can achieve favorable clinical results.It can effectively restore shoulder function.

Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P＜0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.