Development a multiplex panel of AISNPs, multi-allelic InDels, microhaplotypes, and Y-SNP/InDel loci for multiple forensic purposes via the NGS.

Recently, next generation sequencing has shown the promising application value in forensic research. In this study, we constructed a multiplex amplification system of different molecular genetic markers based on the previous selected ancestry informative single nucleotide polymorphisms (SNPs), multi-allelic insertion/deletion (InDel) polymorphisms, microhaplotypes, and Y-chromosomal SNP/InDel loci, and evaluated forensic efficiencies of the system in Chinese Shaanxi Han, Chinese Hui, and Chinese Mongolian groups via the next generation sequencing platform. Ancestry information analyses of Shaanxi Han, Hui, and Mongolian groups revealed that most Mongolian individuals could be differentiated from Shaanxi Hans and Huis based on the selected ancestry informative SNPs. Multi-allelic InDels and microhaplotypes showed the multiple allele variations and possessed relatively high genetic polymorphisms in these three groups, indicating these loci could provide higher forensic efficiencies for individual identification and paternity testing. Based on Y-chromosomal SNPs, different haplogroup distributions were observed among Shaanxi Han, Hui, and Mongolian groups. In conclusion, the self-developed system could be used to simultaneously carry out the individual identification, paternity analysis, mixture deconvolution, forensic ancestry information analysis, and Y-chromosomal haplogroup inference, which could provide more valuable investigative clues in forensic practices.

Developing and population analysis of a new multiplex panel of 18 microhaplotypes and compound markers using next generation sequencing and its application in the Shaanxi Han population.

Compound marker consists of two different types of genetic markers, like deletion/insertion polymorphism and single nucleotide polymorphism in the genomic region of 200 bp, and microhaplotype consists of a series of closely linked single nucleotide polymorphisms in a small DNA segment (<300 bp), which show great potential for human identifications and mixture analyses. In this study, we initially selected 23 novel genetic markers comprising 10 microhaplotypes and 13 compound markers according to previously reported single nucleotide polymorphism or deletion/insertion polymorphism loci. Genetic distributions of these 23 loci in different continental populations showed that they could be used as valuable loci for forensic human identification purpose. Besides, high informativeness values (>0.1) were observed in six loci which could be further employed for forensic ancestry analyses. Finally, 18 loci were successfully developed into a multiplex panel and detected by the next generation sequencing (NGS) technology. Further analyses of these 18 loci in the studied Shaanxi Han population showed that 15 loci exhibited relatively high expected heterozygosities (>0.5). Cumulative power of discrimination (0.999 999 999 99 4835) of these 18 loci revealed that the multiplex panel could also be utilized for human identifications in the studied Shaanxi Han population.

[Distribution of traditional Chinese medicine syndromes of diabetic retinopathy and correlation between symptoms].

To investigate the distribution of traditional Chinese medicine (TCM) Syndromes of diabetic retinopathy and explore the correlation between various symptoms. Based on the literature in databases of China Journal Full-text Database (CNKI), Wanfang database, VIP network, China biomedical literature database (CBM) and PubMed, SPSS 20.0 and IBM SPSS Modeler 14.1 software were used to analyze the location of disease, symptoms, tongue and pulse, and syndrome type distribution through frequency statistics. In addition, association rule algorithm was used to explore the basic rules for underlying symptoms combinations of diabetic retinopathy. A total of 560 articles were retrieved, and a total of 240 articles met the inclusion and exclusion criteria. In these documents, 62 types of TCM syndromes were found, involving 9 820 cases, 201 symptoms, 29 tongue conditions, and 36 pulses conditions; the first two locations for the disease were liver and kidney. Then the association rule analysis of high frequency symptoms was used to dig out 15 groups of latent syndrome, and 3 underlying symptom combinations among high frequency symptoms, tongue conditions and pulse conditions. The results of the study showed that Qi and yin deficiency was most common for the diabetic retinopathy, and the location of the disease was closely related to liver and kidney. In addition, these high-frequency symptoms and tongue conditions, pulse conditions, and underlying symptom combinations can occur as main symptoms at diagnosis, providing reference for us to study the epidemiology of PRO scale of the disease. They can also increase the weight of these symptoms directly as the main symptoms, which can be also used as an alternative entry pool for TCM syndrome diagnostic scale, laying foundation for the construction and optimization of TCM symptom database of diabetic retinopathy.

Refractive change in the adult rabbit eye after corneal relaxation with the femtosecond laser.

BACKGROUND: A new procedure to correct myopia that does not disturb the cornea in the optical zone and avoids injuring the corneal epithelium could be a key advance in corneal refractive surgery. The aim of this study is to observe the refractive change in the adult rabbits undergoing femtosecond laser-assisted multilayer intrastromal ablation in the mid-periphery of the cornea without injury of epithelium. METHOD: The right eyes of 8 New Zealand White adult rabbits were used for the experiments. A 60-kHz femtosecond laser delivery system was used, and three lamellar layers of laser pulses were focused starting at a corneal depth of 180 µm and ending at 90 µm from the surface, with each successive layer placed 45 µm anterior to the previous layer. In the interface of the applanation contact lens cone, a 6-mm diameter aluminum circle was placed at the center to block the laser, limiting ablation to the mid-periphery of the cornea. The laser settings were as follows: spot/line separation, 10 µm; diameter, 8.0 mm; energy for ablating the stroma, 1.3 µJ. An authorefractor was used to assess the manifest refraction. RESULTS: Mean spherical equivalent (SE) (mean ± SD, SD: standard deviation) was significantly increased at postoperative week 1 (1.67 ± 0.26 D, p < 0.0001), month 1 (1.65 ± 0.23 D, p < 0.0001), and month 3 (1.60 ± 0.22 D, p < 0.0001) compared to baseline (0.68 ± 0.27 D). Mean spherical equivalent showed no significant change between postoperative week 1 and month 3 (p = 0.1168). CONCLUSION: Femtosecond laser-assisted multilayer corneal intrastromal ablation in the mid-periphery may cause a consequent hyperopic shift with no refractive regression.

[Clinical observation of lymphangiectasis in conjunctivochalasis cases].

OBJECTIVE: To investigate the relationship between the occurrence and development of conjunctivochalasis and bulbar conjunctival lymphangiectasia. METHODS: Case control study. One hundred cases with conjunctivochalasis treated from January to March 2012 were selected to study, and 100 cases with no conjunctivochalasis as the control group at the same time. To observe bulbar conjunctiva lymphatic duct dilatation using slit lamp microscope, analysis bulbar conjunctiva and fascia images by OCT scanning, and ablate lymphatic of conjunctival tissue for pathologic examine. RESULTS: Twenty-nine eyes of the bulbar conjunctiva lymphangiectasia associated with 100 cases (183 eyes) conjunctivochalasis patients, accounting for 15.84%; 8 eyes of the ball conjunctival lymphatic dilation in control group of 100 cases ( 200 eyes), accounting for 4.00%. The difference between the two groups was statistically significant (χ(2) = 15.36, P < 0.001). OCT scanning showed that lymphangiectasia of the conjunctiva is at the subcutaneous mainly, some in the conjunctival lamina propria. They are border-clear, full-filled fluid, single-lumen or multi lumens, not involving the fascia. The histopathological examination showed that the lamina propria of the bulbar conjunctiva mildly chronic inflammatory changes accompanied by a large number of lymphangiectasia. CONCLUSION: Bulbar conjunctival lymphangiectasia may be one of the reasons for the conjunctivochalasis.

[The clinical observation of conjunctivochalasis crescent conjunctival resection with bipolar coagulation].

OBJECTIVE: To seek safe, effective, economical, simple treatment conjunctivochalasis surgical methods, optimize treatment, evaluation conjunctivochalasis surgical treatment. METHODS: A prospective randomized control study, 60 patients (60 eyes) conjunctivochalasis surgery patients were randomly divided into two groups, one line of bipolar coagulation therapy, another group of crescent conjunctival resection. After comparing the two surgical methods ocular surface disease index (OSDI) points, the degree of relaxation conjunctiva, tear meniscus height, BUT, surgical complications, the operation time to evaluate the two kinds of surgical methods of clinical efficacy. RESULTS: Bipolar coagulation therapy with crescent conjunctival resection in 8 weeks after the OSDI points, loose conjunctiva residual points, tear meniscus, BUT the difference was not statistically significant. 8 weeks after bipolar coagulation complications points lower than the crescent conjunctival resection is low, the difference was statistically significant (t = 4.67, P = 0.029); bipolar coagulation operating time (9.53 ± 3.15) min crescent than conjunctival resection time (18.59 ± 7.68) min short, the difference was statistically significant (t = 13.26, P > 0.01). CONCLUSIONS: Conjunctivochalasis line bipolar coagulation and removal of loose conjunctiva crescent with considerable effect, bipolar coagulation was significantly shorter operative time, a significant reduction in postoperative complications, surgical procedures easier.

Neuroprotective effects of curcumin via autophagy induction in 6-hydroxydopamine Parkinson's models.

Curcumin, a polyphenolic compound extracted from curcuma longa, acts as a nontoxic matter with anti-oxidant and anti-inflammatory effects as well as antiproliferative activities. Here, our research aimed to explore the neuroprotective effects of curcumin both in the 6-hydroxydopamine (6-OHDA)-lesioned rat model of Parkinson's disease (PD) in vivo and 6-OHDA-lesioned PC12 cells in vitro. In vitro, 6-OHDA caused a distinct decrease in cell viability of PC12 cells (150 µM). With the incubation of curcumin (1 µM), 6-OHDA-induced apoptosis was suppressed, increasing the autophagy markers (LC3-II/LC3-I, Beclin-1) and inhibiting phosphor-AKT/AKT, phosphor-mTOR/mTOR. In vivo, curcumin (50 mg/kg) reduced the accumulation of a-synuclein and led to higher parkinsonian disability scores in 6-OHDA-lesioned PD rats, contributing to induction of autophagy through inhibiting AKT/mTOR signal pathway. Moreover, treatment with autophagy inhibitors, such as 3-MA and chloroquine, abolished the neuroprotective effects of curcumin as evidence by compromised autophagy and declined motor behavior in PD rats. In conclusion, the present study demonstrated that curcumin repressed PC12 cell death in vitro and improved parkinsonian disability scores in vivo by inhibiting AKT/mTOR signaling pathway which mediated by autophagy, indicating a potential value of curcumin in the therapeutic intervention of Parkinson's disease.

[The expression of Ki-67, TGF-beta(2) and alpha-SMA after the femtosecond laser-assisted intrastromal scanning in rabbit cornea].

OBJECTIVE: To study how the cornea would express Ki-67, TGF-beta(2) and alpha-SMA and how the cornea would heal when it is scanned intrastromally. METHODS: Twenty New Zealand white rabbits were randomly numbered and used for the experiments. The 60 kHz Intralase femtosecond laser delivery system was used to scan 35 rabbit corneas (of these corneas, 5 right corneas were from 5 rabbits). The unoperated 5 left eyes were used as control. The laser settings were: spot/line separation, 10 microm; diameter, 8.5 mm; energy for scanning the stroma, 1.3 microJ; scanning depth, 135 microm, no edge cuts were performed. Slit lamp was used to observe the cornea at 0.5 h, 1 h, 2 h, and 3 months after surgery. On day 1, 3, 7, 30, 90 after surgery, 6 and 1 scanned corneas at each time point were taken for Western blot and immunocytochemical detection respectively to detect the expression of TNF-alpha, Ki-67, TGF-beta(2) and alpha-SMA. RESULTS: Microbubbles were found in corneal stroma after surgery. The number of microbubbles tended to decrease significantly with time. At 2 h, the cornea turned to be transparent again. During 3-month follow-up, the cornea was always transparent and no haze appeared. Western blot analysis and immunocytochemical detection demonstrated that the expression of Ki-67 began to increase at day 1(0.0670 + or - 0.0008) and reached the peak at day 3 after surgery (0.6923 + or - 0.005). While there were significant difference with the control group at day 90 after surgery (t = 24.12, 57.22, 43.26, 39.78, 18.35;P < 0.05), a gradual decrease in expression of ki-67 could be detected from day 3 to 90. There were no significant change in the expression of TGF-beta(2) (t = 0.933, 0.856, 0.934, 0.970, 1.132) and alpha-SMA (t = 1.126, 1.235, 0.993, 1.175, 1.211) at all time points after surgery (P > 0.05). CONCLUSIONS: After intrastromal scanning, the expression of Ki-67 increases and the keratocyte is activated and proliferated. However, due to no epithelium injured and TGF-beta(2) confined to the epithelium, the activated and proliferated keratocytes could not be transformed into myofibroblasts.

The Advantages of Levodopa-Carbidopa Intestinal Gel for Patients with Advanced Parkinson's Disease: A Systematic Review.

BACKGROUND: Levodopa-carbidopa intestinal gel (LCIG) is a new type of administration that results in steadier levodopa plasma concentrations in advanced Parkinson's disease (PD) patients and effectively reduces poor mobility and dyskinesia. METHODS: Electronic databases were searched up to January 1, 2018. The inclusion criteria for this review were as follows: LCIG vs oral medication in advanced PD patients. RESULTS: Five trials, with a total of 198 patients, met all the inclusion criteria. The quality score of these studies ranged from 3 to 5. Two clinical trials showed that compared with oral medication, LCIG had a better treatment effect on on-time with troublesome dyskinesia (TSD) (p = 0.02) and on-time without TSD (p < 0.00001) in advanced PD patients. In addition, four of the 5 studies showed that the LCIG may have better efficacy than oral medication for improving the scores of the UPDRS, and two studies found that LCIG demonstrated better efficacy for improving the PDQ-39 scores. The video recording results indicated a potential decline in both dyskinesia and the "off" state in LCIG-treated patients. The incidence of adverse events was not significantly different between the LCIG and oral medication groups. CONCLUSION: Compared with oral treatment, LCIG exerts its effectiveness, mostly by reducing the time of on-time with TSD, increasing the time of on-time without TSD and scores of UPDRS and PDQ-39. It is suggesting that LCIG was likely to be a new type of administration used in clinical applications. However, due to methodological flaws, these findings should be viewed with caution, and more RCTs are needed in the field to complement our findings.

[The tear proteomics analysis of conjunctivochalasis].

OBJECTIVE: To analysis tear proteins by using shotgun strategy, and to study pathogenesis mechanism of conjunctivochalasis by comparing tear proteins between the conjunctivochalasis and normal body. METHODS: Tears were obtained from 8 conjunctivochalasis cases and 8 normal controls. Fifteen microliters of tears was collected by microcapillary tubes from each eye. Shotgun strategy was used for tear protein analysis. Trypsin digestion in-solution, separation of peptide mixture by reverse-phase high-pressure liquid chromatograph (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS/MS) identification and bioinformation analysis were used in this study. RESULTS: Three hundred and fifty-six proteins were excised in conjunctivochalasis patients and 352 proteins in the normal controls, and among them 119 proteins were the same. The functions of tear proteins were classified with GOA, which found some apoptosis regulation proteins and apoptosis related proteins and inflammatory response related proteins in conjuntivochalasis but not in normal controls group. Defensin was also found in conjuntivochalasis. CONCLUSION: Shotgun strategy can separate and analysis tear proteins effectively, which provide a new method for tear protein component of conjunctivochalasis. The special component in conjunctivochalasis tear show conjunctivochalasis maybe related to cell apoptosis and inflammatory.

[Epidemiologic study of conjunctivochalasis in populations equal or over 60 years old in Caoyangxincun community of Shanghai, China].

OBJECTIVE: To investigate the prevalence and relative factors of conjunctivochalasis among people over 60 years old in Caoyangxincun community of Shanghai. METHODS: A cross-sectional study based on local residents in this community was carried out from September 2008 to October 2008. Cluster sampling was used in randomly selecting individuals aged > or = 60 years. Slit-lamp ophthalmoscope was used for eye examination and diagnosis. The survey was preceded by a pilot study where operational methods were refined and quality assurance evaluation was carried out. A database was set using SPSS 10.0 software, Chi-square test was used for data analysis. RESULTS: 2110 residents took part in this study, with the response rate 94.85%. 930 cases (1762 eyes) were confirmed as conjunctivochalasis, so the prevalence rate is 44.08%. The prevalence of conjunctivochalasis was increasing with age (chi(2) = 10.44, P < 0.01). 556 cases (943 eyes) were confirmed as degree I, 246 cases (647 eyes) as degree II, 120 cases (162 eyes) as degree III and the rest 8 cases (10 eyes) as degree IV. Several symptoms, such as blurred vision, asthenopia and eye pain were more frequent in patient group than in the normal ones (chi(2) value was 15.44, 20.54 and 19.74, respectively, P < 0.01). In the disease eyes, the conjunctiva usually piled up on the nasal and temporal side (944 eyes, 53.58%), the location of inferior lid margin was usually above corneal limbus (2589 eyes, 61.35%) and inferior lid margin entropion and introversion were always found (7.04% and 6.63%). Abnormal lacrimal river was found in 87.23% of the diseased eyes, more than the non-conjunctivochalasis eyes (chi(2) = 1615.81, P < 0.01). CONCLUSIONS: Conjunctivochalasis is a common eye disease in aged population, with obvious signs and symptoms of ocular surface and lacrimal river impairment. In the local residents, most patients were classified as minimal or moderate degree. The location of inferior lid margin and the tension of inferior lid may be related factor of conjunctivochalasis.

Levodopa/Benserazide PLGA Microsphere Prevents L-Dopa-Induced Dyskinesia via Lower ß-Arrestin2 in 6-Hydroxydopamine Parkinson's Rats.

Prolonged pulsatile administration of Levodopa (L-dopa) can generate L-dopa-induced dyskinesia (LID). Numerous research has reported that continuous dopamine delivery (CDD) was useful in reducing the severity of LID. 6-OHDA lesioned rats were divided into two groups to receive intermittent L-dopa stimulation (L-dopa/benserazide) or Levodopa/benserazide PLGA microsphere (LBPM) for 3 weeks. rAAV (recombinant adeno-associated virus) vector was used to overexpress and ablation of ß-arrestin2. We found that LBPM developed less AIM severity compared with standard L-dopa administration, whereas selective deletion of ß-arrestin2 in striatum neurons dramatically enhanced the severity of dyskinesia by LBPM. On the contrary, the effects of LBPM in terms of ALO AIM were further relieved by ß-arrestin2 overexpression. Furthermore, no significant change in motor behavior was seen either in inhibition or overexpression of ß-arrestin2. In short, our experiments provided evidence that LBPM's prevention of LID behavior was likely due to ß-arrestin2, suggesting that a therapy modulating ß-arrestin2 may offer a more efficient anti-dyskinetic method with a low risk of untoward effects.

Striatal overexpression of ß-arrestin2 counteracts L-dopa-induced dyskinesia in 6-hydroxydopamine lesioned Parkinson's disease rats.

Prolonged administration of Levodopa (L-dopa) therapy can generate L-dopa-induced dyskinesia (LID). Accumulating evidence indicates that hyper-activation of the dopamine D1 receptor (D1R) and the cAMP signaling cascade in the medium spiny neurons (MSNs) of the striatum are involved in LID. Previous studies have shown that striatal ß-arrestin2 overexpression significantly reduces LID severity and have indicated that ß-arrestin2 may play a causal role in the dyskinesia sensitization process. L-dopa-induced changes in the expression of signaling molecules and other proteins in the striatum were examined immunohistochemically and by western blot. A rAAV (recombinant adeno-associated virus) vector was used to overexpress and ablate ß-arrestin2. We found that striatal overexpression of AAV-mediated ß-arrestin2 produced less severe AIMs (abnormal involuntary movements) in response to L-dopa, whereas selective deletion of ß-arrestin2 in the striatal neurons dramatically enhanced the severity of dyskinesia induced by L-dopa. Furthermore, no significant improvements in motor behavior (FFT: forelimb functional test) were seen with the inhibition or overexpression of ß-arrestin2. Finally, overexpression of ß-arrestin2 diminished L-dopa-induced D1R and phosphor-DARPP32/ERK levels. Viral deletion of ß-arrestin2 markedly enhanced the key biochemical markers in the direct pathway. We found that increased availability of ß-arrestin2 ameliorated dyskinesia severity with no influence on the anti-Parkinsonian action of L-dopa, suggesting a promising approach for controlling LID in Parkinson's disease. In addition, overexpression of ß-Arrestin2 prevented the development of LID by inhibiting G protein-dependent D1R and phosphor-DARPP32/ERK signaling in dyskinetic rats.

ß-arrestin2 alleviates L-dopa-induced dyskinesia via lower D1R activity in Parkinson's rats.

The cause of the L-dopa-induced dyskinesia (LID) has been ascribed to G-protein coupled receptor (GPCR) supersensitivity and uncontrolled downstream signaling. It is now supposed that ß-arrestin2 affects GPCR signaling through its ability to scaffold various intracellular molecules. We used the rAAV (recombinant adeno-associated virus) vectors to overexpress and ablation of ß-arrestin2. L-dopa-induced changes in expression of signaling molecules and other proteins in the striatum were examined by western blot and immunohistochemically. Our data demonstrated that via AAV-mediated overexpression of ß-arrestin2 attenuated LID performance in 6-OHDA-lesioned rodent models. ß-arrestin2 suppressed LID behavior without compromising the antiparkinsonian effects of L-dopa. Moreover, we also found that the anti-dyskinetic effect of ß-arrestin2 was reversed by SKF38393, a D1R agonist. On the contrary, the rat knockdown study demonstrated that reduced availability of ß-arrestin2 deteriorated LID performance, which was counteracted by SCH23390, a D1R antagonist. These data not only demonstrate a central role for ß-arrestin2/GPCR signaling in LID, but also show the D1R signal pathway changes occurring in response to dopaminergic denervation and pulsatile administration of L-dopa.