Treatment of Diabetic Nephropathy Based on Theory of Generation and Restriction Among Five Elements / 中国实验方剂学杂志

The theory of generation and restriction among five elements， as one of the basic theories in traditional Chinese medicine （TCM）， reveals that treating disease should focus on the root. Since its first record in Huangdi's Internal Classic （Huang Di Nei Jing）， this theory has been covered in many chapters of Synopsis of the Golden Chamber （Jin Gui Yao Lue） and further developed by physicians of later generations， allowing it to serve as a guide for clinical treatment of various diseases. Diabetic nephropathy （DN） is one of the most common complications of diabetes and also a main risk factor for death and disability by virtue of the long-term disease course and complex symptoms. At present， no specific drug is available in western medicine. Considering the close relationship of its complicated etiology and pathogenesis with the five zang organs， DN treatment should focus not only on the kidney， but also other zang organs. Guided by the theory of generation and restriction among five elements， this article believes that DN mainly results from kidney deficiency combined with spleen deficiency and its dysfunction in regulating the water passage. In addition， the exuberance of heart fire and the failure of liver to govern the free flow of Qi are also responsible for the occurrence of DN. Clinically， the therapeutic methods proposed based on theory of generation and restriction among five elements are recommended for DN treatment after the differentiation of actual manifestations into specific syndromes. Specifically， the method of replenishing Huo to nourish Tu is applicable to DN patients with spleen and kidney yang deficiency， the method of nourishing Shui to moisten Mu to those with liver and kidney yin deficiency， the method of mutual generation between Jin and Shui to those with lung and kidney yin deficiency， the method of banking up Tu to generate Jin to those with lung and spleen Qi deficiency， the method of purging the heart and tonifying the kidney to those with non-interaction between heart and kidney， and the method of banking up Tu to control Shui to those with spleen deficiency and fluid retention. Such timely and effective interventions are conducive to delaying the development of DN to end-stage renal disease （ESRD） and improving the clinical outcomes. This article discusses the application of the theory of generation and restriction among five elements in TCM to DN treatment， aiming to provide a theoretical basis for the future application of such new diagnosis and treatment ideas.

Clinical character and genetic mutation of 64 patients with Gitelman syndrome / 基础医学与临床

Objective To study the clinical and genetic profile of the patients with Gitelman syndrome ( GS ) . Methods We retrospectively analyzed the gene mutation and clinical character of 64 GS patients diagnosed in Pe-king Union Medical College Hospital from 2012 to 2016 .Results The age at diagnosis of these 64 patients ( 39 male,25 female) were (35±14) years.At admission, the serum potassium level of the patients was (2.86± 0.44) mmol/L, serum magnesium level was ( 0.62 ±0.14 ) mmol/L, 24-hour urine potassium was ( 82.27 ± 39.73)mmol/day, 24-hour urine calcium was (0.94±0.83)mmol/day and their average blood pressure was 110/69 mmHg.The gene mutation were divided into four groups including homozygous mutation ( n=5) , compound het-erozygous mutation(n=40), multiple mutations (n=9) and single heterozygous mutation (n=10) group.The compound heterozygous group had higher serum potassium concentration ( P<0.05 ) and the homozygous group had a relatively higher serum magnesium concentration but without significance .A total of 74 distinctly different mutation alleles were identified , of which 24 were new mutation alleles .p.Asp486Asn was a hotspot in our series which was found in 16 patients ( 25.0%) .Conclusions There exists great heterogeneity of gene mutation and clini-cal character in Gitelman syndrome .Patients with compound heterozygous have a relatively milder clinical character.p.Asp486Asn mutation is a hotspot in Chinese patients .

Brain natriuretic peptide and copeptin levels are associated with cardiovascular disease in patients with chronic kidney disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cardiovascular disease (CVD) is the leading cause of death in patients with end-stage renal disease (ESRD). We explored the relationship between CVD, plasma brain natriuretic peptide (BNP) and copeptin in non-dialysis patients with chronic kidney disease (CKD).</p><p><b>METHODS</b>BNP and copeptin were measured using ELISA in 86 non-dialysis patients with different degrees of CKD and in 20 control patients. The effects of BNP, copeptin levels and other biochemical indices on carotid ultrasound echocardiography and CVD history were determined using correlation analysis.</p><p><b>RESULTS</b>BNP and copeptin levels were significantly higher in the CKD group than in the control group. Both indices increased progressively, in parallel with the decline in glomerular filtration rate (GFR). BNP levels were (184.25 ± 65.18) ng/L in early phase CKD, (975.245 ± 354.09) ng/L in middle phase CKD, and (1463.51 ± 614.92) ng/ml in end phase CKD compared with levels of (101.56 ± 42.76) ng/L in the control group (all P < 0.01). Copeptin levels in the middle phase ((20.36 ± 9.47) pmol/L) and end phase groups ((54.26 ± 18.23) pmol/L were significantly higher than in the control group ((9.21 ± 2.64) pmol/L; both P < 0.01). There was no difference in copeptin levels between early phase CKD ((10.09 ± 5.23) pmol/L) and control patients. Stepwise multiple regression analysis identified GFR, intima-media thickness (IMT), left ventricular hypertrophy (LVH), and previous history of CVD as independent risk factors for elevated BNP and copeptin levels.</p><p><b>CONCLUSION</b>BNP and copeptin appear to provide sensitive biological markers for the evaluation of atherosclerosis in non-dialysis patients with CKD.</p>

Transfection of hBcl-2 gene protects the liver against ischemia/reperfusion injury in rats during liver transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of hBcl-2 gene transfer on rat liver against ischemia-reperfusion injury, and explore the feasibility of this approach to reduce ischemia-reperfusion injury in liver transplantation.</p><p><b>METHODS</b>We constructed the replication-deficient recombinant adenoviruses Adv-EGFP and Adv-Bcl-2 and transfected them into 293 cells and packaged into adenovirus particles for amplification and purification. The empty plasmid vector virus was constructed similarly. Male SD rats were randomized into Adv-Bcl-2-transfected group, Adv-EGFP-transfected group, ischemia-reperfusion group, and sham-operated group, and liver allograft transplantation model was established by sleeve method. In the transfected groups, the recombinant viruses were administered by perfusion through the portal vein, and the ischemia-reperfusion and sham-operated groups received no treatment. Real-time quantitative PCR and Western blotting were used to detect the mRNA and protein expressions of bcl-2 in the liver tissue of each group, and at 0, 60 and 180 min after reperfusion, serum AST, LDH, and MDA levels were measured. Histological changes of the liver cells were evaluated by HE staining.</p><p><b>RESULTS</b>Bcl-2 mRNA and protein expressions in Adv-Bcl-2-transfected group, as compared with those in Adv-EGFP-transfected group and control group, were significantly increased (P<0.01); the serum levels of AST, LDH and MDA in Adv-Bcl-2-transfected group were significantly lower than those of Adv-EGFP-transfected group and ischemia-reperfusion group (P<0.05 or 0.01). Compared with the sham-operated group, Adv-Bcl-2 treatment group showed lessened edema and vacuolar degeneration of the liver cells without patches or spots of necrosis. In ischemia-reperfusion and Adv-EGFP group, HE staining revealed hepatic lobular destruction and extensive liver cell swelling, enlargement, vacuolar degeneration, edema and occasional focal necrosis.</p><p><b>CONCLUSION</b>Adv-Bcl-2 transfection can induce the expression of bcl-2 gene to reduce ischemia-reperfusion injury of the liver graft in rats.</p>

Application of immunohistochemical labeling in the diagnosis of hepatobiliary intraepithelial neoplasia and/or cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the best combination of monoclonal antibodies for the diagnosis of hepatobiliary intraepithelial neoplasia and/or cancer.</p><p><b>METHODS</b>CK7, CK20, Villin, CEA, P53 and Ki-67 antigens were detected in the tissues of high-level hepatobiliary intraepithelial neoplasia and cancer by immunohistochemistry and the results were analyzed statistically.</p><p><b>RESULTS</b>Villin was 100% positive in hepatobiliary intraepithelial neoplasia and/or cancer, while 100% negative in the adjacent normal bile duct epithelium. The expression rate of CEA was significantly lower in high-level hepatobiliary intraepithelial neoplasia tissues than in the cancer tissues (P<0.05). Ki-67 indexes were significantly lower in most of the high-level hepatobiliary intraepithelial neoplasia than in the cancer tissue (P<0.01). P53 indexes were also lower in high-level hepatobiliary intraepithelial neoplasia (P<0.01).</p><p><b>CONCLUSION</b>Detection of multiple antigens (CEA, Villin, Ki-67 and P53) provides specific clues to the diagnosis of high-grade hepatobiliary intraepithelial neoplasia and/or cancer.</p>

Expression, genetic and epigenetic alterations of LTF gene in nasopharyngeal carcinoma cell lines / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of LTF mRNA in several nasopharyngeal cancer (NPC) cell lines, and analyze the relationship between the genetic and epigenetic changes and expression of LTF gene.</p><p><b>METHODS</b>The expression level of LTF was detected in NPC cell lines HNE1, HNE2, HNE3, CNE1, CNE2, 5-8F, 6-10B cells and tissues of 15 cases of chronic nasopharyngitis by RT-PCR. The LTF protein level was analyzed by Western blotting in 6-10B cells. Then LOH, mutation and methylation status of LTF was examined by microsatellites analysis, PCR-SSCP, MSP and bisulfite genomic sequencing, respectively.</p><p><b>RESULTS</b>15 chronic nasopharyngitis tissues showed stable LTF expression, while there were weak expression in 6-10B cells and absent expression in remaining detected NPC cell lines. There was a significantly lower LTF expression in chronic nasopharyngitis tissues (Z = -3.738, P = 0.000). No LTF protein expression was observed in 6-10B cells. LOH analysis demonstrated that allele loss of LTF wasn't found in NPC cell lines. LTF mutation was noted in 14.3% (1/7) of NPC cell lines. DNA sequencing confirmed the mutation point in the promoter region (-305 bp to -50 bp) was at -218 bp (del T) of LTF gene in the HNE1 cell line. Methylation of LTF gene was not found in chronic nasopharyngitis. However, methylation of LTF promoter was detected in all NPC cell lines. LTF mRNA expression was increased in 5-8F and 6-10B cell lines after treatment with 5-aza-2-deoxycytidine.</p><p><b>CONCLUSION</b>There is an inactivation of expression of LTF gene in the NPC cell lines. Its molecular mechanism may be related with methylation of promoter region and deletion mutation.</p>

Effects of protein tyrosine kinase within the brainstem nucleus tractus solitarius on the ventilatory responses of peripheral chemoreflex / 生理学报

The aim of the present study was to observe whether protein tyrosine kinase (PTK) within the nucleus tractus solitarius (NTS) was involved in the regulation of ventilatory responses of peripheral chemoreflex. The experiments were performed on anesthetized, immobilized and artificially ventilated rabbits. Peripheral chemoreflex was elicited by ventilating the animal with 10% O2-balance 90% N2. Changes in the peak amplitude and frequency of integrated phrenic nerve activity were observed. The ventilatory responses of peripheral chemoreflex following 0.1 microl microinjection within the NTS of either PTK inhibitor genistein (10 mol/L), AMPA glutamate receptor inhibitor CNQX (10 mol/L),or inactive PTK inhibitor daidzein (10 mol/L) were recorded. The results are as follows: Both genistein and CNQX attenuated the ventilatory responses of peripheral chemoreflex, while no changes occurred following daidzein. The amplitude of integrated phrenic nerve discharge and the phrenic burst frequency were decreased by (-21.77+/-6.93)% and (-24.70+/-7.61)% respectively after administration of genistein. CNQX resulted in similar decreases in the amplitude of phrenic nerve discharge (-27.13+/-7.63)% and the burst frequency (-21.34+/-4.88)%. In addition, the inhibitory effects of CNQX and genistein were the same whether they were applied alone or one after another, indicating that they had no cooperative effects. The results obtained suggest that PTK within the NTS regulates the peripheral chemoreflex control of respiration and that this regulation of PTK may be mediated through the phosphorylation of AMPA receptors in NTS neurons.

The locus coeruleus modulates the inspiratory inhibition induced by electrical stimulation of the Bötzinger complex / 生理学报

Experiments were done on urethane anesthetized adult rabbits. Long-train electrical stimulation was delivered to the Bötzinger complex (Böt.C) to observe the changes in the peak amplitude of integrated phrenic nerve activity. Then, a long-train electrical stimulation was delivered to the locus coeruleus (LC) or monosodium glutamate was microinjected into the LC . Within a certain period of time, another long-train electrical stimulation was delivered to the Böt.C to observe the responses of phrenic nerve activity. We investigated whether the LC could modulate the inspiratory inhibition induced by electrical stimulation of the Böt.C. The results are as follows: (1) Within a certain period of time after a long-train electrical stimulation applied at the LC, the inspiratory inhibition produced by electrical stimulation at the Böt.C was significantly attenuated. Comparing with the control stimulation that was only delivered at Böt.C without pre-stimulation of the LC, the inspiratory inhibition was decreased by (28.78+/-19.49)%. (2) Similarly, after chemical stimulation of the LC with microinjection of monosodium glutamate, the inspiratory inhibition produced by electrical stimulation of Böt.C was also significantly attenuated [decreased by (19.18+/-8.06)%]. The results obtained suggest that the LC plays a role in the modulation of the inspiratory inhibition of Böt.C stimulation.

Sustained electrical stimulation of Bötzinger complex on phrenic nerve discharges in rabbits / 中国应用生理学杂志

<p><b>AIM</b>To observe the effects of sustained electrical stimulation at Bötzinger complex (Böt. c) on phrenic nerve discharges.</p><p><b>METHODS</b>Sustained electrical stimulation (10--50 microA, 40-100 Hz, 0.3 ms, for 15-30 s) of Böt. C on 30 urethane anaesthetized, vagotomized, paralyzed and artificially ventilated rabbits.</p><p><b>RESULTS</b>Sustained electrical stimulation of Bot. C produced the inhibition or "inspiratory off-switch" of phrenic discharge during the stimulation. The inhibition of the phrenic discharges showed intensity and frequency dependence. Habituation was shown during the stimulation, showing the magnitude of the phrenic nerve discharge increased gradually. Post-stimulus rebound exhibited upon the cessation of the stimulation, showing the magnitude of the phrenic activity increased significantly. Short-term memory was shown in the habituation of the phrenic activity.</p><p><b>CONCLUSION</b>Non-associative learning is involved in the central control of respiratory modulation in the Böt. C and synaptic plasticity may exist in the respiratory neurons of Böt. C.</p>

Selective myelo-protection by MDR1 and MnSOD genes regulated by a specific promoter / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the specific protection of myeloid cells from chemotherapeutic agents and radiation.</p><p><b>METHODS</b>The recombinant retroviral vectors containing MDR1 gene and MnSOD gene regulated by APN myeloid promoter were constructed and introduced into myeloblastic cell line KG1a and hepatoma cell line BEL7402. The resistance of the cells to antitumor drugs and radiation were analyzed by cell survival assay. In vivo, the murine bone marrow cells were isolated and infected by the retroviral particles, which were transplanted into recipient mouse treated with paclitaxel or X-ray. The murine white blood cell (WBC) was counted in order to assay the effects of MDR1 or MnSOD gene on hematopoiesis in the course of chemotherapy and radiotherapy.</p><p><b>RESULTS</b>The resistance to chemotherapeutic agents such as cochicine, Vp-16, vincristine, doxorubcin and paclitaxel were elevated markedly by 10.6, 10.4, 11.2, 4.2 and 14.2 folds in KG1a cell line transduced with MDR1 gene. The resistance to radiation increased 3.7 folds at the dose of 10 Gy compared with parental cells in KGla cell line transduced with MnSOD gene derived by APN promoter. In contrast, the chemosensitivity and the radiosensitivity showed no significant change in BEL 7402 cell line transduced with MDR1 gene and MnSOD gene. In vivo, the WBC counts in the mouse introduced with MDR1 gene or MnSOD gene were higher than those in the control mouse (P < 0.01).</p><p><b>CONCLUSION</b>The expression of MDR1 gene and MnSOD gene regulated by APN myeloid promoter is effective on myelo-specific protection without enhancing the resistance of tumor cells in vitro. The hematopoiesis can be reconstituted in vivo during anticancer drug or radiation treatment. This study may provide experimental evidence and new clues for myeloprotection of cancer patients being treated with chemotherapy and/or radiotherapy.</p>

The effect of estrogen benzoate on normal human breast tissue implanted into athymic nude mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the proliferating effect of estradiol benzoate on normal human breast tissue.</p><p><b>METHODS</b>Three specimens of normal human breast tissue were implanted into nine 9-10-week-old intact female athymic nude mice which were randomly divided into group A, B and C. Each specimen was divided into three parts, and each part was implanted into mice of each group of three (A, B and C) respectively. Two weeks later, part of xenografts was taken out from group A as control group, and then group A, B and C were injected muscularly with 3, 6, 9 micrograms estradiol benzoate once daily, respectively. After therapy for 28 days, the grafts were taken out and expression of proliferating cell nuclear antigen (PCNA), estrogen receptor (ER) and proliferating receptor (PR) in grafts were examined using immunohistochemical method.</p><p><b>RESULTS</b>Compared with control group, PCNA was increased in group B and C (P < 0.05), but in group A had no significant change. ER was gradually down regulated by 3, 6 and 9 micrograms estradiol benzoate (P < 0.05); however, PR was increasingly up regulated (P < 0.05).</p><p><b>CONCLUSIONS</b>Certain dosage of estradiol benzoate has proliferating effect on normal human breast tissue.</p>

Clinical rehabilitation research on 90 infants with cerebral palsy / 中国康复理论与实践

@#ObjectiveTo investigate the effect of rehabilitative treatment on different groups of infants with cerebral palsy in different age, sickness extent and rehabilitative period.MethodsBy the prospective method, 90 patients from 3 to 36 months old were randomly divided into three groups including 3 to 12 months, 13 to 24 months and 25 to 36 months with 30 cases each. Every 10 cases in light, moderate and severe degrees were given rehabilitative care for 2 months and 3 months respectively. The personal ADL of every case was evaluated at the initial and the final state. The compared score was analyzed with T test.ResultsComparing the groups of different ages, there were prominent differences of treatment under the same sickness, among the light, moderate and severe degrees(P<0.01). The rehabilitative treatment effect of 3 months was better than that of 2 months. Furthermore the effect of the group of 3-12 months was better than that of 25-36 months.ConclusionsThe rehabilitative care is prominently effective in the light and moderate groups , the former is better and within 3 months is better than 2 months. The younger infant is, the better the rehabilitative treatment is.