Objective To construct a suppression subtractive hybridization (SSH) library of human renal cell carcinoma. Methods Poly A + RNA was isolated from human renal cell carcinoma (RCC) tissues and matched normal kidney tissues, respectively. Then double strand cDNA were synthesized and restricted by Rsa I.RCC cDNA were divided into two groups and ligated with either adaptor 1 or adaptor 2.After RCC cDNA hybridized with normal kidney cDNA twice and underwent nested PCR,the PCR products were cloned into pT Adv vector and transformed E.coli TOP10F′.Some positive clones randomly picked up were digested and sequenced. Results The SSH library contained 414 positive clones. Random analysis of 265 clones with enzyme restriction showed that 246 clones contained cDNA fragments which were distributed mainly between 300～900 bp. Among 40 arbitrary clones derived from the above 246 clones, No.170 clone is a previously unknown sequence and the other 39 clones were derived from 35 known genes. Conclusions The quality of the SSH library of human RCC is reliable and its construction would lay the foundation for further screening differentially expressed genes in RCC.

Objective To investigate the influence of Guhong injection (GHI) on ATPase activity and inflammatory response after cerebral ischemia/reperfusion (I/R) injury in rats, and evaluate its protective effects on cerebral I/R injury. Methods Seventy-two male Sprague-Dawley (SD) rats were divided into sham group, I/R group, nimodipine group, and the low-dose (2.5 mL/kg, GHI-L), medium-dose (5.0 mL/kg, GHI-M), and high-dose (10.0 mL/kg, GHI-H) of GHI groups according to the random number table method, with 12 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by the intraluminal suture method to prepare the model of focal cerebral ischemia, and reperfusion was performed after 1.5 hours of occluding the middle cerebral artery; the sham group had the same operation process except inserting the nylon thread. The injection of drug in various drug-treated groups was carried out via a tail vein at 0, 12, 24 hours after the onset of reperfusion, while the sham group and I/R group received the same amount of normal saline. At 12 hours after last drug administration, the scores of neurological deficit symptoms were evaluated; the cerebral infarction was observed by triphenyl tetrazolium chloride (TTC) staining; the Na+-K+-ATPase and Ca2+-ATPase activities in the brain tissue were measured by phosphorus determination; the contents of interleukin-6 (IL-6), monocyte chemotactic factor-1 (MCP-1), nitric oxide (NO) in serum were detected by enzyme linked immunosorbent assay (ELISA). Results Compared with the sham group, the neurological function score was significantly decreased, the cerebral infarction was serious, the activities of ATPase was obviously decreased, and the levels of serum inflammatory factors were significantly increased in I/R group. Compared with the I/R group, the neurological function scores were significantly increased in GHI-L group, GHI-M group, GHI-H group and nimodipine group (9.03±0.63, 10.54±2.55, 12.33±1.87, 12.06±1.89 vs. 8.17±1.05, all P < 0.05), the volumes of cerebral infarction were obviously reduced [(18.51±1.80)%, (15.98±1.34)%, (8.61±1.16)%, (8.09±0.96)% vs. (26.52±2.07)%, all P < 0.01], the activities of ATPase were significantly increased [Na+-K+-ATPase (μmol·mg-1·h-1):5.10±0.30, 5.34±0.26, 6.19±0.17, 5.86±0.31 vs. 3.98±0.35, Ca2+-ATPase (μmol·mg-1·h-1): 3.68±0.44, 4.43±0.29, 5.03±0.27, 4.17±0.30 vs. 1.87±0.46, all P < 0.01], and the levels of serum inflammatory factors were decreased obviously [IL-6 (ng/L): 51.61±5.55, 43.88±4.05, 39.71±2.22, 41.28±2.66 vs. 60.11±6.61, MCP-1 (ng/L): 227.82±7.07, 201.58±13.10, 177.23±10.46, 126.80±8.49 vs. 296.01±12.85, NO (μmol/L): 54.48±3.23, 46.84±2.69, 41.15±2.80, 48.62±2.34 vs. 65.25±3.88, all P < 0.05]. Conclusions GHI not only can improve the energy metabolism of brain tissue in a dose-dependent manner, but also inhibit the inflammatory cascade of damage after cerebral I/R in rats, which might be its protective mechanism on cerebral ischemia injury.

Megalencephalic leukoencephalopathy with subcortical cysts (MLC, OMIN: 604004) caused by mutations in the MLC1 gene, is an rare autosomal recessive disorder. More patients are with infancy and young children onset, whereas adult cases are rare. Only 2 patients from 1 family have been reported in domestic adult cases. Now a 58-year-old female MLC patient is reported. The clinical manifestations of the patient included large head circumference, slow responses, walking difficulties, seizures and paroxysmal loss of consciousness. The result of whole exome sequencing revealed a homozygous insertion mutation c.920_943dup in the MLC1 gene. The mutation in this patient has not been reported in the Human Gene Mutation Database.

Intestinal microbes play an important role in human health. The development of various clinical diseases, such as obesity, diabetes and cardiovascular disease, is closely related to the imbalance of intestinal microflora. With the development of high-throughput sequencing technology, there has been a breakthrough in the understanding of intestinal microorganism. The interaction between intestinal epithelial cells and intestinal microbes has become one of the hotspots and difficulties of current research. Because of the constraints of ethical review and experimental cost, people are more interested in the development of interaction models between the intestinal microflora and the host cells. In this paper, interaction models between intestinal microflora and host cells, and its working principle and application prospect are reviewed, hoping to provide new techniques and new ideas for studying functions of intestinal microbes.