Understanding autism spectrum disorders with animal models: applications, insights, and perspectives.

Autism spectrum disorder (ASD) is typically characterized by common deficits in social skills and repetitive/stereotyped behaviors. It is widely accepted that genetic and environmental factors solely or in combination cause ASD. However, the underlying pathogenic mechanism is unclear due to its highly heterogeneous nature. To better understand the pathogenesis of ASD, various animal models have been generated, which can be generally divided into genetic, environment-induced, and idiopathic animal models. In this review, we summarize the common animals used for ASD study and then discuss the applications, clinical insights, as well as challenges and prospects of current ASD animal models.

Combined treatment with vitamin K2 and PTH enhanced bone formation in ovariectomized rats and increased differentiation of osteoblast in vitro.

Osteoporosis is accompanied by insufficient osteogenic capacity. Several lines of evidence suggested that solutions to enhance osteoblastogenesis were important strategies for osteoporotic bone defect repair. This study investigated the effect of combined treatment with vitamin K2 and PTH on bone formation in calvarial bone defect in osteoporotic rats and its influence on osteoblast in vitro. Bilateral ovariectomy was used in SPF Sprague Dawley rats to generate an osteoporosis model. Subsequently, a calvarial defect model was established and all osteoporotic rats were randomly assigned to the following groups: control, VK (vitamin K2, 30 mg/kg everyday), PTH (recombinant human PTH (1-34), 60 µg/kg, three times a week) or VK + PTH (vitamin K2, 30 mg/kg everyday plus PTH, 60 µg/kg three times a week) for 8 weeks. In vitro, bone marrow-derived stem cells (BMSCs) were cultured and treated with vitamin K2, PTH or vitamin K2+PTH. ALP staining and western blot were performed to observe the influence of combined treatment on BMSCs. Bone formation within calvarial defect were assessed by serum γ-carboxylated osteocalcin (Gla-OC), micro-CT, histological and immunofluorescent labeling. In this study, combined treatment of PTH and vitamin K2 showed positive effects on preventing bone loss in femurs in OVX rats. Combined treatment increased serum Gla-OC and promoted bone formation in osteoporotic calvarial bone defects. Immunohistochemistry showed that OCN and RUNX2 were more highly expressed in the VK + PTH group than in the control groups. In vitro studies results suggested that combined treatment with PTH and vitamin K2 increased expression of ALP, BMP2 and RUNX2 in BMSCs. Our data suggested that the combination of vitamin K2 and PTH increased differentiation of osteoblast and had a synergistic effect on bone formation in osteoporotic calvarial bone defect.

[Stiletto needle and needle-knife for influence of gravity index in treating knee osteoarthritis].

OBJECTIVE: To explore stiletto needle and needle-knife for influence of double sufficient weight in treating knee osteoarthritis patients. METHODS: One hundred and thirteen early and medium term knee osteoarthritis patients were randomly divided into three groups, including stiletto needle group(38 cases), needle-knife group (38 cases) and voltaren group (37 cases). In stiletto needle group, there were 13 males and 25 females with an average of(55.87±7.72) years old, treated by stiletto needle once a week, and 2 weeks were a course; there were 11 males and 27 females in needle-knife group with an average of(57.11±7.07) years old, treated by acupotome once a week, and 2 weeks were a course; there were 12 males and 25 females in voltaren group with an average age of(57.62±8.08) years old, treated by votalin emulsion smearing 3 to 5 cm on painful area of knee joint, three times a day for 2 weeks; 36 patients in normal group, including 11 males and 25 females with a mean age of (55.28±7.55) years old, treated with nothing. Gravitational four lattice used to measure bipedal back and forth load before and after treatment in further observe weight-bearing situation among three groups, d value, which was the distance from center of gravity to original point, was measured as a obvervational index, JOA score was used to evaluate clinical effect. RESULTS: Five patients were fall out, including 2 patients in stiletto needle group, 2 patients in needle-knife group and 1 patient in voltaren group. Other 108 patients were followed-up from 28 to 35 d with an average of 30 d, and without untoward effect. There was significant difference in d value between treatment group and control group at 1 month after treatment(P<0.05); there was significant difference in d value among treatment group at 1 month after treatment(P>0.05), and d value was decreased before treatment than that of after treatment. There was no significant difference in JOA score among treatment group after treatment at 1 month(P<0.05); while had significant difference between groups(P>0.05) after treatment at 1 month. CONCLUSIONS: Stiletto needle, needle-knife and voltaren for the treatment of knee osteoarthritis at early and middle stage, center of gravity is closer to original point, and weight-bearing is improved. It is a easy method of four lattice to measure change of weight-bearing in KOA patients after treatment.

Nickel N-heterocyclic carbene catalyzed C-C bond formation: a new route to aryl ketones.

A novel nickel N-heterocyclic carbene catalyzed cross-coupling reaction of aryl aldehydes with boronic esters for the synthesis of aryl ketones was developed. This reaction provides a mild, practical method toward aryl ketones, which are versatile intermediates and building blocks in organic synthesis.

[Lethal effects of Trichoderma longibrachiatum on Heterodera avenae].

The lethal effect of the conidia suspension of Trichoderma longibrachiatum against second stage juveniles of Heterodera avenae was observed under microscopic conditions and studied in vitro to preliminarily determine the potential and mechanism of the conidia suspension of T. longibrachiatum against H. avenae. Microscopic observation results showed that the conidia suspension of T. longibrachiatum adhered to or parasitized on the surface of second stage juveniles, even some parasitized nematodes were deformed at the initial stage. Later, the second stage juveniles were wrapped and the integuments were penetrated by a large number of hyphae germinated from the conidia suspension of T. longibrachiatum. Even the body of dead second stage juveniles was completely lysed. In vitro studies showed that different concentrations (1.5 x 10(5)-1.5 x 10(7) cfu x mL(-1)) of T. longibrachiatum (conidia suspension) had strong lethal and parasitic effects on the second stage juveniles of H. avenae, and significant differences existed among the treatments with different concentrations of T. longibrachiatum. In addition, the lethal and parasitic effects increased with increasing the T. longibrachiatum concentration. The mortality and corrected mortality of the second stage juveniles treated with the concentrations of 1.5 x 10(7) cfu x mL(-1) were 91.3% and 90.4% after 72 hours, and the second stage juveniles were parasitized by 88.7% after 14 days. Therefore, the conidia suspension of T. longibrachiatum had a great lethal effect on H. avenae, and the strain could be considered as a potential bio-control agent against H. avenae.

[Case-control study on the treatment of patients with scapular muscle fasciitis by Pizhen].

OBJECTIVE: To evaluate and analyze the mechanism of Pizhen for the treatment of patients with scapular muscle fasciitis. METHODS: One hundred and sixteen patients were divided into two groups according to random number table method, 58 cases in the treatment group and 58 cases in the control group. After treatment, 109 patients were followed up, and 7 patients lost follow-up (2 cases in the treatment group and 5 patients in the control group). In the treatment group, there were 15 males and 41 females, with a mean age of (28.02 +/- 4.85) years; the course of disease ranged from 0.25 to 3 years; and the patients were treated by Pizhen once a week,and 2 weeks were one treatment course. In the control group, there were 19 males and 34 females, with a mean age of (27.23 +/- 4.54) years old; the course of disease ranged from 0.25 to 1 year; and the patients were treated by acupuncture massage instrument, 20 minutes at a time,once a day for 2 weeks. Soft tissue displacement and banana area were tested by a soft tissue tension tester, pressure value was measured by pressure measuring instrument, and VAS was recorded by pain visual analog scale record. RESULTS: There were significant differences in tissue displacement, banana area, pressure value and VAS between two groups before and after treatment. The tissue displacement and pressure value in the treatment group were higher than those in the control group,while banana area and VAS were lower than those in the control group. In the treatment group, 30 patients healed, 13 patients got marked improvement, 12 good and 1 poor;while 16 patients healed, 19 patients got marked improvement,11 good and 7 poor in the control group. The therapeutic effects in the treatment group was better than that in the control group. CONCLUSION: Pizhen is an effective method to treat pain in patients with scapular muscle fasciitis by relaxing tendon of soft tissue around pain point of scapular and relaxing compress and stimulation of nerves.

[Clinical evaluation of stiletto needle for pain of knee osteoarthritis].

OBJECTIVE: To objectively evaluate the clinical efficacy of stiletto needle for pain of knee osteoarthri tis (KOA), and analyze its function mechanism. METHODS: Seventy-six cases of KOA (76 knees) were selected. Under the guide of Jingjin theory in TCM, stiletto needle was applied at pain point of Jingjin in extra-articular area to have a loose solution effect, 1 to 3 points were selected each time, 1 to 2 times of treatment were required. The results of tenderness measurement instrument was adopted as main evaluation index of joint pain, and all data of evaluation indices before and after the treatment were statistical analyzed. RESULTS: There were significant differences in visual analogue scale (VAS) score, tenderness score, HSS function score and movement range of joint before and after the treatment (all P < 0.05). The effective rate of stiletto needle therapy was 89.5%. There was apparent regression trend between VAS score and tenderness score with Y (VAS) = 7.841-1.569 X (tenderness score) as its regressive equation. CONCLUSION: The stiletto needle therapy is an effective method to relieve the pain of knee osteoarthritis, and its clinical efficacy evaluation could be more objective and digital with tenderness measurement instrument.

[Distribution and physicochemical properties of aquatic colloids in the Yangtze estuarine and coastal ecosystem].

Cross-flow ultrafiltration (CFUF) technique was applied for isolating colloids from waters in the Yangtze estuarine and coastal ecosystem. The stability and physicochemical properties of colloids were analyzed quantitatively. Colloidal size and Zeta-potential at XP and WSK station decreased with the increasing pH, and reached the point of zero charge at pH < 2 and pH 3.8, respectively. The colloidal organic carbon (COC) concentration in the collected samples was 7.7-35.7 micromol x L(-1), occupying 7.1% -41.7% of the total dissolved organic carbon (DOC). Three-dimensional excitation/emission matrix fluorescence spectroscopy (3DEEM) was used to analyze the samples and the results showed tryptophan-like fluorescence peaks, UV fulvic-like fluorescence peaks and humic-like fluorescence peaks. Concentrations of colloidal Na, Mg, K, Ca, Co, Cr, Cu, Fe, Li, Mn and Ni accounted for 0.33%, 5.7%, 0.975%, 1%, 7.2%, 7%, 11.9%, 15.7%, 5.5%, 10.5% and 11.3% of the dissolved phase in average, and the binding capacity of Co, Cr, Cu, Fe, Li, Mn, Ni with COC was generally higher than that of Na, Mg, K, Ca. There were significant correlations between colloidal size and salinity, between DOC and salinity, and between UOC and salinity, but no linear relationship between COC and salinity was found. The total concentration of trace metals (Co, Cr, Cu, Fe, Li, Mn, Ni) and COC showed a linear relationship.

[Parasitic and lethal effects of Trichoderma longibrachiatum on Heterodera avenae: microscopic observation and bioassay].

A laboratory experiment was conducted to study the parasitic and lethal effects of Trichoderma longibrachiatum conidia suspension on Heterodera avenae cysts. Different concentrations (1.5 x 10(5)-1.5 x 10(7) cfu x mL(-1)) of T. longibrachiatum conidia suspension had strong parasitic and lethal effects on H. avenae cysts, and the effects differed significantly among the different concentrations. When treated with the T. longibrachiatum conidia suspension at a concentration of 1.5 x 10(7) cfu x mL(-1), 96.7% of the H. avenae cysts were parasitized by the conidia at the 18th day, and the hatching rate of the cysts was inhibited by 91.2% at the 22nd day. The microscopic observation showed that at the initial parasitic stage, T. longibrachiatum conidia suspension adhered or parasitized on the cyst surface, germinated a large number of hyphae, and grew on the cyst surface, making the development of cyst embryo stopped and the contents in cysts flocculated, and even, some cysts started to deform, and small dark brown vacuoles formed on the cyst surface. At the later parasitic stage, the cysts were penetrated by dense mycelium, cysts were broken, their contents exosmosed, and the mycelium on the integument of some cysts produced conidiophores, on which, conidium were adhered or parasitized. It was considered that T. longibrachiatum could be used as a potential high-efficient bioagent to control the occurrence and damage of H. avenae.

[Case-control study on the treatment of pain in patients with knee osteoarthritis by Pizhen].

OBJECTIVE: To evaluate and analyze mechanism of Pizhen for the treatment of pain in patients with knee osteoarthritis. METHODS: Eighty-five patients (85 knees) were divided into two groups according to random number table method. Among 35 patients in the experimental group, 6 patients were male and 29 patients were female, 1 patient lost follow-up, the average age was (56.65 +/- 4.89) years, and the patients were treated with Pizhen. Among 50 patients in the control group,there were 6 males and 44 females, the average age was (57.54 +/- 4.91) years, and the patients were treated with intra-articular injection with sodium hyaluronate. The VAS score and the data of algometer from the most obvious tenderness around the knee were used to evaluate the clinical effects. RESULTS: There were significant differences of scores in the experimental group before and after treatment (P<0.01). There were significant differences of scores in the control group before and after treatment (P<0.01). After treatment, there were no statistical differences of VAS score and the data of algometer between experimental and control groups ( P>0.01). There was negative correlation between VAS score and the data of algometer. CONCLUSION: The Pizhen is an effective method to treat pain in patients with knee osteoarthritis by relaxing tendon of soft tissue around pain point of knee joint, and relaxing compress and stimulation of nerves.

[Case-control study on stiletto needle and Dichofenac Diethylammon for the treatment of knee osteoarthritis pain].

OBJECTIVE: To investigate therapeutic effects of stiletto needle and Dichofenac Diethylammon in treating knee osteoarthritis pain at early and medium-term. METHODS: Sixty-seven patients with early and medium-term knee osteoarthritis were randomly divided into two groups, 34 cases in stiletto needle group (treatment group) and 33 cases in Dichofenac Diethylammon (control group). Among them, 60 patients finished treatment and followed-up for 1 months (7 cases fall off totally, 4 cases in treatment group and 3 cases in control group). In treatment group, there were 5 males and 25 females with a mean age of (55.90 +/- 9.34) years (ranged, 40 to 68 years), the course of disease for 0.25 to 1 year, mean (0.87 +/- 0.34) years, treated by stiletto needle once a week, and 3 weeks were a course. In control group, there were 6 males and 24 females with a mean age of (58.67 +/- 7.39) years (ranged, 40 to 70 years), the course of disease for 0.25 to 2 years, mean (0.93 +/- 0.60) years, treated by Dichofenac Diethylammon, three times a day for 3 weeks. Soft tissue displacement and banana area were tested by soft tissue tension tester, pressure value were measured by pressure measuring instrument, VAS score were recorded by pain visual analog scale record, HSS scoring and effects were used to evaluated and statistical analyzed. RESULTS: There were significant differences in tissue displacement, banana area, pressure value, VAS scoring and HSS scoring between two groups before and after treatment, and tissue displacement, pressure value, HSS scoring in treatment group were higher than control group, while banana area and VAS scoring were lower than control group. According to HSS scoring and VAS scoring, 3 cases were healed, 8 cases were markedly improved, 18 cases were effective and 1 case was effectiveless in treatment group;while 1 case was healed, 4 cases were markedly improved, 20 cases were effective and 5 cases were effectiveless in control group. There was no significant meaning in therapeutic effects. CONCLUSION: Stiletto needle for knee osteoarthritis pain can effectively reduce high tension of local soft tissue ,alleviate symptom of pain, and improve knee function.

[Grading treatment of hip osteoarthritis].

OBJECTIVE: To investigate therapeutic effects of intra-articular injection of sodium hyaluronate, arthroscopy and total hip replacement for the treatment of different severity hip osteoarthritis. METHODS: Eighty-nine patients were treated with the three methods from June 2004 to August 2008. There were 33 males and 56 females, ranging in age from 30 to 72 years, averaged 51.8 years. The disease course ranged from 3 to 360 months, with a mean of 58.7 months. All the patients had hip pain and limited movement of hip joint. The patients were treated according to ISOA score and Kellgren-Lawrance (K-L) grades of hip osteoarthritis. The preoperative and postoperative Harris scores were compared. RESULTS: After the operation, 83 patients were followed up, and the duration ranged from 3 to 58 months. The Harris scores results: the patients with slight hip osteoarthritis and K-L grade I got an averaged preoperative score of (70.50 +/- 4.62) and postoperative score of (91.75 +/- 4.01); the patients with slight hip osteoarthritis and K-L grade II got an averaged preoperative score of (70.15 +/- 3.79) and postoperative score of (82.18 +/- 3.92). The patients with moderate hip osteoarthritis and K-L grade III got an averaged preoperative score of (57.12 +/- 2.19) and postoperative score of (89.94 +/- 1.13) score. The patients with serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (48.63 +/- 2.21) and postoperative score of (90.76 +/- 1.14); the patients with very serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (42.67 +/- 3.17) and postoperative score of (91.97 +/- 2.24); the patients with extremely serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (36.46 +/- 2.96) and postoperative score of (82.69 +/- 1.19). There were significant differences in scores of all groups before and after treatment. CONCLUSION: The hip osteoarthritis should be treated according to its serious degree such as slight, moderate and severe, as well as based on its X-ray classification, which is the best method to get satisfactory results.

[Study on midterm follow-up results of arthroscopic debridement for knee osteoarthritis].

OBJECTIVE: To evaluate the effect of arthroscopic debridement for knee osteoarthritis. METHODS: The medical records of 56 patients (56 knees) who underwent arthroscopic debridement procedures for knee osteoarthritis from 2000 to 2004 were reviewed. Among the patients, 15 patients were male and 41 patients were female,ranged in age from 39 to 76 years, with an average of (55.23 +/- 10.26) years. The duration of the disease ranged from 1 to 25 months, with an average of 5.75 months. The chief symptoms were pain, swelling and dysfunction of the knees. According to Kellgren-Lawrence classification grade, 17 patients were grade I ,39 patients were grade II. Lysholm score was used to evaluate the postoperative effects, and the statistics analysis was carried out with SPSS 13.0. RESULTS: All the patients were followed up more than 5 years. Three patients underwent total knee replacement respectively at 5.5, 7.8 and 8.3 years after the arthroscopic debridement, the average Lysholm score of other 53 patients increased from (42.40 +/- 6.78) preoperatively to (75.53 +/- 8.23) postoperatively. There were significant difference between preoperative score and postoperative score (t = -22.62, P < 0.01). CONCLUSION: The good midterm follow-up results of arthroscopic debridement for knee osteoarthritis are related to the appropriate indications selections, limited debridement, good patient education, systemic rehabilitation and chondroprotective agents.

[Expression of DPC4/Smad4, p21wafI, and p16 in human pancreatic cancer].

BACKGROUND & OBJECTIVE: DPC4/Smad4 inactivation was detected in almost half of the pancreatic carcinomas result in the loss of inhibition of the tumor cell proliferation. p21wafI is the downstream target gene of Smad4 while DPC4 and p16 may synergistically play a role in the development of pancreatic carcinoma. By studying the expressions of DPC4/Smad4, p21wafI, and p16. This study was designed to explore the mutual relationship among them and the possible mechanism in human pancreatic carcinoma. METHODS: Immunohistochemistry was used to detect the expression of Smad4, p21wafI, and p16 in fifty-six samples of paraffin embedded human pancreatic cancer tissue, and in situ hybridization, immunohistochemistry, Western blotting technique were used to detect the expression of DPC4/Smad4 in five human pancreatic adenocarcinoma cell lines. RESULTS: The positive rate of Smad4, p21wafI, and p16 in paraffin embedded human pancreatic cancer tissue was 58.93%, 48.21%, and 42.86%, respectively, whereas the positive rate of these proteins in matched normal tissue was 89.29%, 87.5%, and 76.79% respectively. Three out of five pancreatic adenocarcinoma cell lines (P3, P4, and P7) were positive for DPC4/Smad4 with in situ hybridization, immunohistochemistry and Western blotting, while the other two lines were all negative. There is statistically significant difference between cancer and normal tissue (P < 0.05). In pancreatic adenocarcinomas, the expression of Smad4 was related to that of p21wafI (P < 0.05), and so was the expression of Smad4 to that of p16 (P < 0.05). But no correlation was found between p21wafI and p16 (P > 0.05). CONCLUSION: The expression of Smad4, p21wafI, and p16 significantly decreased in pancreatic cancer compared with normal tissue. The decreased expression of the proteins may play an important role in the development of pancreatic cancer.

[Alterations of DPC4/SMAD4/MADH4 gene detected in paraffin-embedded tissues of human pancreatic carcinomas].

OBJECTIVE: To demonstrate the alterations of DPC4/SMAD4/MADH4 gene in paraffin-embedded tissues of pancreatic carcinomas. METHODS: Forty-six cases of resected specimens containing carcinomatous tissue and normal pancreatic tissue were analysed for possible DPC4 gene mutations by polymerase chain reaction (PCR)and single-strand conformation polymorphism (SSCP). The DNA sequencing technique was applied to determine the patterns of gene mutation in the PCR-SSCP positive cases. Fifty-six cases of pancreatic carcinoma along with the specimens corresponding normal pancreatic tissues were studied by in situ hybridization (ISH) and immunohistochemistry (IHC) techniques for gene expression in mRNA and protein level. RESULTS: The homozygous deletion rate of exon 1, 2, 3, 4, 8, 11 of DPC4 gene in pancreatic carcinoma was 28.26%, while the mutation rate of DPC4 gene was 21.74%. In these tumors, there were 3 cases of nonsense mutation, 5 cases of missense mutation, 1 case of deletion and missense mutation, 1 case of insertion mutation. Positive rates of SMAD4 in carcinomatous tissues detected by the ISH and IHC were 53.57% and 58.93% respectively, whereas they were 91.07% and 89.29% in the matched normal tissue respectively. There were significant difference between cancer and normal tissue (P < 0.05). Thrity-two cases were positive of DPC4/SMAD4 with all methods mentioned above, the coincident rate was 87.50% (28/32). The coincidence between gene detection and ISH of SMAD4 was 87.50%, and it was 96.88% between gene detection and IHC of SMAD4. Of all 56 cases, the coincidence of the positive rates of SMAD4 detected by ISH and IHC was 91.07%. No significant difference among the positive rates of DPC4/SMAD4 as detected by the three different techniques (P > 0.05). CONCLUSIONS: The main mechanisms of inactivation of DPC4 gene in pancreatic carcinoma are homozygous deletion and mutation. The product of DPC4 expression is significantly decreased in cancer group compared with the normal tissues. As a tumor suppressor gene, DPC4 alteration is an important molecular event in pancreatic carcinoma, and probably plays a crucial role in cancer development and progression.